Vous êtes sur la page 1sur 384

FOOD SCIENCE AND TECHNOLOGY

FOOD CHEMISTRY

No part of this digital document may be reproduced, stored in a retrieval system or transmitted in any form or
by any means. The publisher has taken reasonable care in the preparation of this digital document, but makes no
expressed or implied warranty of any kind and assumes no responsibility for any errors or omissions. No
liability is assumed for incidental or consequential damages in connection with or arising out of information
contained herein. This digital document is sold with the clear understanding that the publisher is not engaged in
rendering legal, medical or any other professional services.
FOOD SCIENCE AND TECHNOLOGY

Additional books in this series can be found on Nova‘s website


under the Series tab.

Additional E-books in this series can be found on Nova‘s website


under the E-book tab.

FOOD AND BEVERAGE CONSUMPTION


AND HEALTH

Additional books in this series can be found on Nova‘s website


under the Series tab.

Additional E-books in this series can be found on Nova‘s website


under the E-book tab.
FOOD SCIENCE AND TECHNOLOGY

FOOD CHEMISTRY

DONGFENG WANG
HONG LIN
JIANQIAN KAN
LINWEI LIU
XIAOXING ZENG
AND
SHENGRONG SHEN
EDITORS

New York
Copyright © 2012 by Nova Science Publishers, Inc.

All rights reserved. No part of this book may be reproduced, stored in a retrieval system or
transmitted in any form or by any means: electronic, electrostatic, magnetic, tape, mechanical
photocopying, recording or otherwise without the written permission of the Publisher.

For permission to use material from this book please contact us:
Telephone 631-231-7269; Fax 631-231-8175
Web Site: http://www.novapublishers.com

NOTICE TO THE READER

The Publisher has taken reasonable care in the preparation of this book, but makes no expressed or
implied warranty of any kind and assumes no responsibility for any errors or omissions. No
liability is assumed for incidental or consequential damages in connection with or arising out of
information contained in this book. The Publisher shall not be liable for any special,
consequential, or exemplary damages resulting, in whole or in part, from the readers‘ use of, or
reliance upon, this material. Any parts of this book based on government reports are so indicated
and copyright is claimed for those parts to the extent applicable to compilations of such works.

Independent verification should be sought for any data, advice or recommendations contained in
this book. In addition, no responsibility is assumed by the publisher for any injury and/or damage
to persons or property arising from any methods, products, instructions, ideas or otherwise
contained in this publication.

This publication is designed to provide accurate and authoritative information with regard to the
subject matter covered herein. It is sold with the clear understanding that the Publisher is not
engaged in rendering legal or any other professional services. If legal or any other expert
assistance is required, the services of a competent person should be sought. FROM A
DECLARATION OF PARTICIPANTS JOINTLY ADOPTED BY A COMMITTEE OF THE
AMERICAN BAR ASSOCIATION AND A COMMITTEE OF PUBLISHERS.

Additional color graphics may be available in the e-book version of this book.

Library of Congress Cataloging-in-Publication Data

Food chemistry / editors, Dongfeng Wang ... [et al.].


p. cm.
Includes index.
ISBN: B
1. Food--Analysis. 2. Food--Composition. I. Wang, Dongfeng.
TX531.F555 2011
664'.07--dc23
2011042522

Published by Nova Science Publishers, Inc. † New York


CONTENTS

Preface vii
Contributors ix
About the Editors xi
Chapter 1 Introduction 1
Dongfeng Wang
Chapter 2 Water 9
Jianqian Kan and Guoqing Huang
Chapter 3 Carbohydrates 35
Dongfeng Wang, Jipeng Sun, Guoqing Huang,
Xiaolin Zhou and Liping Sun
Chapter 4 Lipids 107
Shengrong Shen, Dongfeng Wang and Undurti N. Das
Chapter 5 Proteins 137
Hong Lin, Lisha Wu and Shuhui Wang
Chapter 6 Vitamins 191
Yibin Zhou, Dongfeng Wang and Ping Dong
Chapter 7 Minerals 223
Dongfeng Wang, Lina Yu, Haiyan Li, Bin Zhang,
Shuhui Wang and Xingguo Liang
Chapter 8 Food Flavors 247
Xiaoxiong Zeng and Guaoqing Huang
Chapter 9 Food Additives 273
Linwei Liu and Shiyuan Dong
Chapter 10 Toxicants in Foods 305
Wang Dongfeng, Guoqing Huang and Shuhui Wang
Index 353
PREFACE

Foods consist of a large quantity of compounds, of which, some are original from plant or
animal materials, some are new ones generated during processing or preservation, some are
intentionally added by manufacturers, and some are contaminants produced during
processing, preservation or packaging. These compounds undergo various changes during
processing and storage and it is hence necessary to understand the effects of processing or
storage on these compounds so as to enhance the nutrition, palatability and safety of foods.
The purpose of Food Chemistry is to elucidate the structure, physicochemical properties,
nutrition and safety of major food constituents and their changes occurred during processing
and storage. Due to the extreme importance, Food Chemistry has been accepted as a major
fundamental course for food-related majors.
Though food chemistry has a history of more than 200 years, it developed into a
relatively independent system in the late 1960‘s. Since then, the United States, Japan,
Germany and other countries published several authoritative food chemistry textbooks,
including Latest Food Chemistry edited by Hayashi Junzo and Kitamura Mitsuo (Japan),
Food Chemistry by Sakurai Yoshito (Japan), Food Chemistry by Owen R. Fennema (United
States), Food Chemistry by Belitz HD (Germany), Food Chemistry by Zhang Wang (China),
and Food Chemistry by Dongfeng Wang (China). Of the works, the publications edited by
Fennema and Belitz HD have been widely chosen by university students as textbook.
However, the two books contain too many contents and part of them overlaps with those
stated in Biochemistry and Organic Chemistry. Besides, the two books are too expensive for
readers in developing countries.
Hence, there is an urgent demand to publish a simplified Food Chemistry textbook that
most university students can afford, which is the case of this book. This book presents the
chemistry and properties of the six essential nutrients contained in foods, including water,
carbohydrates, lipids, proteins, vitamins and minerals, and their changes occurred during food
processing and storage. In addition, this book also deals with the chemistry and properties of
flavors, food additives and toxic substances in foods. This book is simplified and cheaper
than previously published books without reducing its academic level, and reflects the latest
advances in food chemistry. This work can be used as a textbook by university students and
especially suitable for students in developing countries and non-English speaking countries
for bilingual delivery.
The authors would like to thank the postgraduates of the Laboratory of Food Chemistry
and Nutrition of Ocean University of China, including Mei Ding, Yan Li, Lu Yu, Xingya Li,
viii Dongfeng Wang, Hong Lin, Jianqian Kan et al.

Xiang Gao, Wen Zhou, Zhe Xu, Min Wang, Mengqi Li, and Chunsheng Li, for assistance in
literature collection and typesetting, and Ocean University of China for funding the
publication.
CONTRIBUTORS

Undurti N Das
Jawaharlal Nehru Technological University, Kakinada-533 003, India

Liping Sun
College of Chemistry and Engineering, Kunming University of Science and Technology,
Yunnan Province, China

Jipeng Sun
Third Institute of Oceanography State Oceanic Administration, Xiamen, China

Lina Yu
Shandong Peanut Research Institute, Qingdao, China

Xiaoling Zhou
Medical College of Shantou University, Shantou, Guangdong Province, China

Bin Zhang
School of Food and Pharmacy & Medical School, Zhejiang Ocean University, Zhushang
City, Zhejiang Province, China

Haiyan Li
College of Food Science and Engineering, Ocean University of China, Qingdao, China

Banping Wang
College of Food Science and Engineering, Ocean University of China, Qingdao, China

Xingguo Liang
College of Food Science and Engineering, Ocean University of China, Qingdao, China
ABOUT THE EDITORS

Dongfeng Wang is a professor of the College of Food Science and Engineering at the
Ocean University of China. He has published many books related to food chemistry as editor-
in-chief, including Food Chemistry (2007), Advanced Food Chemistry (2009), Chemistry of
Toxic Substances in Foods (2005), Technology of Experiment & Study of Tea Biochemistry
(1997), Experiments on Food Quality & Food Safety (2004) and Technology of Experiments
on Food Science and Engineering (2007). He has published over 120 original papers that
reflect his research interests in food chemistry, tea biochemistry, carbohydrate chemistry, and
preservation. He has received many teaching & academic honors, including The Second Prize
for Advanced Science and Technology of China in 2010, The First Prize for Advanced
Science and Technology from Ministry of Education of the People‘s Republic of China in
2009, Distinguished Teacher Awards from Shandong Province of China in 2006, and Award
for Young Scientists from Anhui Province in 2000. Professor Wang received his BS degree of
agriculture in 1982 from Anhui Agricultural College (Anhui, China), the MS degree of tea
biochemistry in 1988 from Zhejiang Agricultural University (Zhejiang, China), and the PhD
degree of inorganic biochemistry of food in 1999 from University of Science and Technology
of China (Hefei, China).

Jianquan Kan is a professor of the College of Food Science at Southwest University of


China. He is the editor-in-chief of many books related to food chemistry, including The
Practical Chemistry of Oil and Fat (1997), Food Chemistry (2002, revised in 2006 and 2008),
Advanced Food Chemistry (2011), An Introduction to Food Safety (2009), Food Analysis
(2011) and Experimental Methods (2011). He is the author or corresponding author of over
140 original papers covering food chemistry, food analysis and nutrition. He has been
honored the Second-Class Prize of Chongqing Science and Technology Advancement (2009)
and the Second Chongqing Academic and Technological Leader (2008). Professor Kan
received the BS degree of chemistry from Nanchong Normal College (Sichuan, China) in
1986, the MS degree of Product Processing and Storage from Southwest Agricultural
University (Chongqing, China) in 1992, and the PhD degree of Product Processing and
Storage from Southwest Agricultural University (Chongqing, China) in 2003.

Lingwei Liu is a professor of the College of Food Science & Engineering in Northwest
A&F University, Yangling, ShaanXi, China. He has done intensive researches related to food
chemistry, food analysis, nutrition and food safety. Professor Liu received the BS degree of
food science from Northwest Agriculture University (ShaanXi, China) in 1982 and the PhD
xii Dongfeng Wang, Hong Lin, Jianqian Kan et al.

degree of food science from Northwest Agriculture & Forest University (ShaanXi, China) in
1995.

Hong Lin received the BS degree in 1984, the MS degree in 1990, and the PhD degree
in 1998 in seafood science from the Ocean University of China. Professor Lin is a famous
expert in the seafood safety field and his research area covers novel marine organism-derived
chemical and biological hazard discovery, quality control during seafood processing, and fast
hazard detection method development. Professor Lin has been granted 4 invention patents,
published more than 100 original articles, and edited 4 academic books, including Seafood
Safety (2010), Aquatic Nutrition and Safety (2007), Effective Use of Aquatic Resources
(2007), and Fish Preservation Technologies (2000).

Xiaoxiong Zeng received the BS degree from Hunan Agricultural University in 1985,
the MS degree from Zhejiang Agricultural University in 1988, and the PhD degree from
Shizuoka University (Shizuoka, Japan) in 2000. Dr. Zeng is now a professor of the College of
Food Science and Technology, Nanjing Agricultural University, China. He is one of the
authors or corresponding author of over 100 original papers related to food chemistry, food
biotechnology and glycobiology.

Shengrong Shen is a professor of the School of Biosystems Engineering and Food


Science of Zhejiang University. Professor Shen was granted the PhD degree by the
Department of Biophysics of Zhejiang University in 1997. His research area includes
structural analysis of such bioactive compounds as lipids, fatty acids, and polyphenols. He
has published more than 100 original papers concerning food chemistry, food safety and
applied nutrition in the latest 10 years. Besides, professor Shen has published 5 academic
books related to food and health, tea biochemistry and food chemistry.

Yibin Zhou is a professor of the Department of Food Science and Engineering at Anhui
Agricultural University, Anhui, China. He had edited Food Chemistry (in Chinese) as an
assistant, and is the author or corresponding author of over 40 original papers on
carbohydrates, food engineering, and biotechnology.

Guoqing Huang is a lecturer of the College of Food Science and Engineering, Qingdao
Agricultural University, Qingdao, China.

Shiyuan Dong is a lecturer of the College of Food Science and Engineering, Ocean
University of China, Qingdao, China.

Shuhui Wang is a PhD candidate in Biosystems Engineering Department College of


Agriculture - Ginn College of Engineering, Aubum University, Auburn, AL 36849-5417,
USA
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 1

INTRODUCTION

Dongfeng Wang
College of Food Science and Engineering, Ocean University of China, Qingdao, China

ABSTRACT
Food Chemistry is a fundamental discipline for students, engineers, and
professionals engaged in the food industry. This chapter provides an overview of this
discipline, including its definition, purpose, development, and its role in food science and
engineering.

1.1. Food Chemistry and Its History

1.1.1. What Is Food Chemistry


Nutrients refer to the indispensable substance that provides nourishment essential for
maintenance of life, growth and development of human being. The human body needs a lot of
nutrients. Based on chemical structure, the nutrients can be divided into six major categories,
including water, carbohydrates, proteins, lipids, vitamins and minerals.
A minor difference between terms foodstuff and food should be noted first. Foodstuff
refers to materials containing nutrients; while foods are materials that have been processed
from foodstuff (ranging from simple cleaning to a modern factory processing) in order to
meet people‘s nutritional and sensory requirements. In another word, a food shall be
characterized by both nutrition and sensory satisfaction.
The nutritional compositions of foods can be determined easily, but sensory satisfaction
is a much complex issue and is related to the color, texture, and shape, flavor of foods in
addition to the cultural background and dietary habits of consumers.
The chemical compositions of foods are very complex (Figure 1-1). Of the components,
some are intrinsic in animal or plant materials, some are generated during processing and
storage, some are intentionally added by manufacturers, some are contaminants originated
from the environment or microorganisms, and some are migrated from packing materials of
2 Dongfeng Wang

food. The purpose of Food Chemistry is to elucidate the structure, physical and chemical
properties, nutritional value as well as safety of these components, their changes undergone
during storage and processing, and the effects of these changes on food nutrition and
palatability. The knowledge is of great importance in improving food quality, developing new
food resources, evolving food processing and storage technologies, upgrading food packaging
materials, and increasing food safety and quality.

Water
Carbohydrates
Proteins
Lipids
Minerals
Natural
Vitamins
Pigments
Hormones
Flavor components
Toxic substances

Food components

Natural additives
Food Addtives
Synthetic additives
Unnatural
From processing
Contaminants
From environmental pollution
Figure 1-1. Composition of foods.

Food Chemistry is a comprehensive discipline and partially overlaps with chemistry,


biochemistry, physical chemistry, botany, zoology, food nutrition, food safety, polymer
chemistry, environmental chemistry, toxicology, molecular biology, and many other subjects.
Food Chemistry associates the most closely with chemistry and biochemistry and it is the
extension of the two subjects to the food area. However, the subjects have different contents
and focuses. The chemistry subject deals mainly with the composition, property, and
reactions of molecules, biochemistry focuses on the reactions and changes of various
components in organisms under suitable or moderately suitable conditions, while food
chemistry is interested in the changes of components occurred in such unsuitable conditions
as freezing, heating, and drying, their interactions during these processes, and the effects of
these changes on the nutrition, safety, and sensory properties (such as color, flavor, taste, and
shape) of foods.

1.1.2. History of Food Chemistry


It is a short time since Food Chemistry is accepted as an independent subject. However,
the researches and reports related to this subject have been started since the last 1700s. Many
components were separated from foods by chemists and botanists at that time and Researches
on the Chemistry of Food by Justus von Liebig in 1847 is recognized as the first book related
to food chemistry.
Introduction 3

As the trading of foods between regions and countries increased, both consumers and
manufacturers had urgent needs on the information of water contents and the presence non-
food components in foods. Meanwhile, driven by the rapid development of analysis measures,
the desire to understand the natural characteristic of foods also grew. In 1860, German
scholars Hanneberg W. and Stohman F. invented a method for the simultaneous
determination of water, crude fat, ash, and nitrogen contents. Several years later, diets
containing solely proteins, lipids, and carbohydrates were found insufficient for maintaining
life.
In 1900s, with the advancement of analytical techniques and the biochemistry subject and
the rapid development of the food industry, requirements on new food processing
technologies and prolonged storage life emerged, which drove the quick development of food
chemistry. During this period, a growing number of researches papers were published and the
quantity of related journals increased significantly as well, including Archives of Biochemistry
and Biophysics (initiated in 1942), Journal of Agricultural and Food Chemistry (initiated in
1953) and Food Chemistry (initiated in 1966). Due to the emergency of increasing deep and
systematic publications, Food Chemistry gradually developed into an independent subject.
Chinese scholars Yanbin Xia and Ruijin Yang divide the history of Food Chemistry into
four stages.
Stage one: Many natural components were separated from plants and animals and were
identified, including lactic acid, citric acid, malic acid, and tartaric acid. The knowledge was
not systematic yet and was reported mainly by chemists.
Stage two: In the early 1900s (1820 ~ 1850), food chemistry developed quickly along
with the development of agricultural chemistry and gained much importance in Europe.
Specialized food chemistry laboratories were established and many professional journals
related to food chemistry were issued. Meanwhile, adulteration became a serious issue and
the need for impurity determination propelled the development of food chemistry. In this
stage, Justus von Liebig invented an optimized method for quantitative analysis of organic
substances and published Researches on the Chemistry of Food in 1847.
Stage three: In the middle 1900s century, the British scientist Arthur Hill Hassall reported
the microscopic images of pure and adulterated foods and food chemistry came into the
microanalysis time. In 1871, Jean Baptis M.D.M. proposed that diets containing only
proteins, carbohydrates and lipids were insufficient to sustain human‘s life. The interests on
the nutritional requirements further accelerated the development of food chemistry. Until the
first half of the 20th century, the majority of components in foods were identified and the
number of literatures related to chemistry food increased markedly. Food chemistry then
turned to be a mature and independent subject in mid-20th century.
Stage four: Food chemistry is now in the fourth stage. With the rapid development of
society, economy, science and technology, and the improvement of living standards,
consumers raise higher requirements on food security, nutrition, palatability, and
convenience. Meanwhile, to realize the transformation from traditional to scaled,
standardized, and modernized processing of foods, more and more new technologies,
materials, and equipment are used, which markedly drive the rapid development of food
chemistry. Besides, the advancement of basic chemistry, biochemistry, instrumental analysis
and other related subjects guarantee the rapid development of food chemistry. Food chemistry
has become a most important subject for food scientists [1, 2].
4 Dongfeng Wang

1.1.3. Food Chemistry Textbooks


A series of food chemistry textbooks were published between 1976 to 1985, including
Latest Food Chemistry by Hayashi Junzo and Kitamura Mitsuo (Japan), Food Chemistry by
Sakurai Yoshito (Japan), Food Chemistry by Owen R. Fennema (United States), and Food
Chemistry by Belitz HD (Germany), in which, the works of Fennenma and Belitz HD
contributed a lot to the development of food chemistry and has been widely chosen by
university students as textbook. Food Chemistry has been chosen as a fundamental course for
food related majors.

1.2. The Role of Food Chemistry in Food Science and Engineering

Foodstuff undergoes various chemical and biochemical reactions during storage,


transport, and processing. These reactions might yield products that are either beneficial to
food nutrition and palatability or harmful to consumers. The knowledge of food chemistry is
hence of extreme importance, because the purpose of this subject is to elucidate the changes
of various food components occurred during storage, transport, and processing and the effects
of these changes on food quality. In recent years, the control of composition, property,
structure, and interaction of various food components, the chemical nature of the nutrition and
palatability of complex food systems, and the exploitation of new food resources constitute
the new contents of food chemistry. With the development of science and technologies and
the extension of other fundamental subjects to the food industry, more and more toxic and
harmful chemicals in foods are identified and food chemistry has turned to be the theoretical
foundation for guaranteeing food quality and safety. Food chemistry plays an important role
in food science and engineering and is developing quickly.

1.2.1. Role of Food Chemistry in Technology Advancement


Nutrition, healthcare, safety, and enjoyment are the four fundamental attributes of foods
required by the modern food industry. The theories and application research results of food
chemistry are guiding the healthy and sustainable development of the food industry (Table 1-
1). Practice has proved that, no the theoretical guidance of food industry, no the ever growing
modern food industry.

Table 1-1. Impact of food chemistry on technological advancement of the


food industry [3, 4]

Food Industry Application


Flour improving; starch modification; new edible materials exploitation;
high-fructose syrup; food enzymes; molecular basis of food nutrition; new
Basic food sweetener and natural additive development; new oligosaccharide
industry production; oil modification; vegetable protein isolates; functional peptides
production; microbial polysaccharides and single cell protein development;
development and utilization of wild, marine, an edible drug resources, etc.
Storage and Chemical peeling; color protection; texture control; vitamin retention; de-
processing of astringency and debittering; coating and waxing; chemical preservation;
fruits and controlled atmosphere storage; bioactive packaging; enzyme-assisted
vegetables juicing, filtration and clarification; chemical preservation, etc.
Introduction 5

Table 1-1. (Continued)

Food Industry Application


Post-slaughter processing; juice preservation and tenderization; color
protection and development; enhancement of the emulsifying capacity,
Storage and
gelling capacity, and viscoelasticity of meat; frozen denaturation of
processing of
proteins; fresh meat packaging in supermarket; production and application
meats
of fumigation agent; artificial meat production; comprehensive utilization of
viscera, etc
Instant dissolution; ingredient floating and/or sinking inhibition; protein
beverage stabilization; water treatment; juice stabilization; juice color
Beverage protection; flavor enhancement; alcohol degree decrease; beer clarification;
industry beer foamability and bitterness improvement; chemical nature and
prevention of beer non-biological stability; off-flavor elimination; juice de-
astringency; soybean odor elimination, etc.
Yoghurt and juice milk stabilization; chymosin substitute development;
Dairy industry
whey utilization; nutrition fortification of diary products; etc.
Baking High-efficiency leavening agent development; crispness improvement;
industry bread color and texture modification; aging and mildewing inhibition; etc.
Edible oils Lipid refinement; lipid modification; development and utilization of DHA,
and fats EPA, and MCT; food emulsifier and anti-oxidant development; oil
industry absorption reduction of fried foods; etc.
Condiments Meat soup production; nucleotide-type flavor enhancers; organic iodine-
industry supplemented salt; etc.
Fermented Post-processing of fermented foods; flavor changes during post-
food industry fermentation; comprehensive utilization of biomass and residues; etc.
Source identification of exogenous toxicants and their prevention;
Food safety
identification of endogenous toxicants and their elimination; etc
Food Formulation of inspection standards; rapid analysis; biosensor development;
inspection fingerprint preparation of products; etc.

Due to the rapid development of food chemistry, some important reactions, including the
Millard reaction, caramelization, lipid auto-oxidation, starch gelatinization and aging,
polysaccharide hydrolysis and modification, protein hydrolysis and denaturation, pigment
discoloration, vitamin degradation, metal-catalyzed reactions, enzyme-catalyzed reactions, fat
hydrolysis and transesterification, lipid thermo-oxidative decomposition and polymerization,
flavor compound changes, action mechanisms of food additives, generation of harmful
ingredients as well as postharvest physiology, are identified in foods. The knowledge on these
reactions greatly enhances the development of the food industry.

1.2.2. Role of Food Chemistry in Human Nutrition and Health


It has been more than two centuries since proteins, carbohydrates and lipids were
identified as the three major nutrients for human. The two most important attributes of foods
are to provide consumers with nutrition and sensory satisfaction. One of the objectives of
food chemistry is to investigate the nutrition and flavor composition in food materials and
processed foods and the interactions of the components occurred during processing and
storage and effects of these interactions on food nutrition and palatability. The modern food
6 Dongfeng Wang

chemistry should not only ensure the healthcare and enjoyment attributes of food
components, but also guide consumers on rational diet selection. The concept of nutrition has
evolved significantly due to social development and the change of the healthy status of
consumers. How to reduce the incidences of diet-related diseases, such as cardio-
cerebrovascular diseases, cancers, and diabetes, has turned to be a new major task of food
industry. In addition to the healthcare attribute, foods should also provide desirable flavors so
that consumers enjoy the eating process. The emergence of biotechnologies and new food
processing technologies guarantees the safety of foods.
Contamination of foods by pollutants is currently a worldwide concern due to global
environmental deterioration. The analysis and identification of trace and ultramicro
substances are of vital importance to the nutrition value and the control of toxicants of foods.
The development of food chemistry has been associated with the healthy status and
civilization level of human.

1.3. Research Methods of Food Chemistry

Each type of food contains a large number of components and is thereby a much complex
system. Hence, the research methods of food chemistry are quite different from those of
common chemistry subjects. In food chemistry, the knowledge on the chemical composition,
physicochemical properties, and changes of food components must be associated with the
nutrition, enjoyment, and safety of foods. The experimental design of food chemistry should
reveal the complex composition of food systems and the changes of the nutrition value,
enjoyment, and safety of foods during processing and storage. The interactions between food
components and their changes occurred during storage and processing (such as ultra-high
pressure, high temperature, freezing, presence or absence of oxygen) are extremely complex.
Hence, many researches are carried out in simplified and stimulated models, which must be
then verified in real food systems.
The experiments of food chemistry include mainly physicochemical experiments and
sensory evaluation experiments. Physicochemical experiments reveal the composition of
foods and the structures of the components, including nutrients, toxicants, and flavors; while
sensory experiments evaluate the texture, flavor, and color changes of foods through visual
inspection.
Foods or food materials undergo a series of changes during storage, transport, processing,
and sales. The changes include: enzymatic and chemical reactions in raw and fresh materials;
changes caused by water activity variance; component decomposition, polymerization, and
denaturation under violent conditions (high temperature, high pressure, mechanical actions);
oxidation induced by oxygen or other oxidants; photochemical reactions; and migration of
packaging materials to foods. Of the changes, non-enzymatic browning, lipid oxidation and
hydrolysis, protein hydrolysis and denaturation, protein cross-linking, oligosaccharide and
polysaccharide hydrolysis, and change of the presence form of natural pigments and their
degradation, are the most important reactions for the food industry. Of the reactions, some are
desired, but some are unexpected and must be avoided during processing (Table 1-2). The
mechanisms and control of these reactions constitute the key contents of food chemistry.
Introduction 7

Table 1-2. Part reactions occurred during food processing and storage and their
influences on foods [3, 4]

Reaction Examples Influence on foods


Nonenzymic Color development in Desired or undesired color, smell, taste; loss of
browning bakery foods nutrition; harmful ingredients.
Oxidation Oxidation of lipids, Change color; desired flavor or off-odors and
vitamins, an phenols toxicants
Hydrolysis Hydrolysis of lipids, Increased soluble solids content; texture
proteins, and carbohydrates changes; desired color, flavor, taste, and
nutrition; toxicity loss of certain components

Isomerization cis-trans isomerization of Discoloration; formation or loss of certain


lipids functions
Polymerization Foam and insoluble brown Discoloration; loss of nutrition; off-odor
precipitate forming in development; toxicants formation
frying
Protein Egg white coagulation; Improved nutrition; toxicity loss of certain
denaturation enzyme inactivation components

The research fruits and methods of food chemistry have been widely absorbed by the
food industry and greatly promote the development of the food industry. In the last decades,
some new subjects and research areas, such as structural chemistry, free radical chemistry,
membrane separation, edible package, microencapsulation, extrusion, superfine comminution,
bioactive packaging, supercritical extraction, molecular distillation, membrane catalysis, bio-
reactor, toxicant chemistry of foods, molecular nutrition, and nutria-genomics, have been
established. These new technologies and subjects will undoubtedly facilitate the rapid
development of the food industry, which in turn benefits the improvement of the food
chemistry subject.

REFERENCES
[1] Wang, DF. Food Chemistry.1st edition. Beijing: Chemistry Industry Press; 2007
[2] Damodaran, S; Parkin, KL; Fennema, OR. Fennema’s Food Chemistry. 4th edition.
New York: CRC Press; 2007.
[3] Kan, JQ. Food Chemistry. 1st edition. Beijing: China Agricultural University Press;
2002.
[4] Wang, Z. Food Chemistry. 1st edition. Beijing: China Light Industry Press; 2005.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 2

WATER

Jianqian Kan1 and Guoqing Huang2


1
College of Food Science, Southwest University, Chongqing, China
2
College of Food Science and Engineering, Qingdao Agricultural University,
Qingdao, China

ABSTRACT
Water is an important component in many foods. Its content and occurrence status
significantly affect the flavor, texture, and stability of foods. This chapter deals with the
various physical and chemical properties of water and ice and the interactions with other
components in foods. Water occurs in multiple states due to interactions with solutes and
the interactions significantly affect the bioavailability of water to chemical reactions and
microorganisms.
To distinguish the differences between water content and its bioavailability, the term
water activity (aw) is proposed and its application in food stability predication are
detailed. The relationship between water content and aw can be presented by moisture
sorption isotherm (MSI), which is very useful in designing the concentration and
dehydration processes of foods.
In addition to aw, molecular mobility (Mm) has also been proposed to predict food
stability. Its definition and its effect on food stability are also a concern of this chapter.
Water is a predominant constituent in many foods (Table 1). Water in proper amount,
location, and orientation profoundly influences the structure, appearance, and taste of
foods and their susceptibility to spoilage.
Because medium water supports chemical reactions and water is a reactant in
hydrolytic processes, the removal of water from foods retards many reactions and inhibits
the growth of microorganisms, thus improving the shelf lives of a number of foods.
Through physical interaction with proteins, polysaccharides, lipids and salts, water
contributes significantly to food texture.
Water is essential to life: as an important governor of body temperature, as a solvent,
as a carrier of nutrients and waste products, as a reactant and reaction medium, as a
lubricant and plasticizer, as a stabilizer of biopolymer conformation, as a likely facilitator
of the dynamic behavior of macromolecules, including their catalytic (enzymatic)
properties, and in other ways yet unknown.
10 Jianqian Kan and Guoqing Huang

Table 1. Water contents of some foods [1]

Water content Water content


Food Food
(%) (%)
Pork, raw, composite of lean 53~60 Cereal flour 10~13
Beef, raw, retail cuts 50~70 Honey 20
Chicken, all classes, raw Avocado, bananas, peas
74 74~80
meat without skin (green)
Beets, broccoli, carrots,
Fish, muscle proteins 65~81 80~85
potatoes
Asparagus, beans
bananas 75 (green), cabbage, 90~95
cauliflower, lettuce
Berries, cherries, pears 80~85 Bread 35~45
Apples, peaches, oranges,
85~90 Biscuits 3~8
grapefruit
Rhubarb, strawberries,
90~95 Tea 3~7
tomatos
Butter, margarine 15 Edible oil 0
Milk powder 4

1. PHYSICAL AND CHEMICAL PROPERTIES OF WATER AND ICE


1.1. The Water Molecule and Its Association

1.1.1. The Water Molecule


The water molecule is comprised of two hydrogen atoms interacting with the two sp3
bonding orbitals of oxygen, forming two covalent σ bonds. A schematic orbital model of a
water molecule is shown in Figure 1.a and the appropriate van der Waals radii are shown in
Figure 1.b.

Figure 1. Schematic model of a single HOH molecule: (a) sp3 configuration, and (b) van der Waals radii
for a HOH molecule in the vapor state [1].
Water 11

In the vapor state, the bond angle of an isolated water molecule is 104.5°. The O-H
internuclear distance is 0.96 Å and the van der Waals radii for oxygen and hydrogen are 1.40
and 1.2 Å respectively.

1.1.2. Association of Water Molecules


Each water molecule has an equal number of hydrogen-bond donors and receptor sites
and is able to hydrogen-bond with a maximum of four water molecules. The resulting
tetrahedral arrangement is shown in Figure 2. The two unshared electron pairs (n-electrons or
sp3 orbitals) of oxygen act as H-bond acceptor sites and the H-O bonding orbitals act as
hydrogen bond donor site. The dissociation energy of this hydrogen bond is about 11-
25kJ/mol.
As mentioned above, each water molecule can hydrogen bond with at most four water
molecules and the resultant three-dimensional structure is quite stable. This structure is quite
different from those formed by other small molecules that also involved in hydrogen bonding
(such as NH3 and HF). Ammonia has three hydrogen-bond donors and one hydrogen-bond
receptor, while HF has one hydrogen and three receptor sites. Both the two chemicals do not
have equal numbers of donor and receptor sites and therefore can form only two dimensional
hydrogen-bonded networks. The above mentioned polarization of H-O bonds is transferred
via hydrogen bonds and extends over several bonds. Therefore, the dipole moment of a
complex consisting of increasing numbers of water molecules is higher as more molecules
become associated and is certainly much higher than the dipole moment of a single molecule.
Proton transport takes place along the H-bridges. It is actually the jump of a proton from one
water molecule to a neighboring water molecule. In this way a hydrate H3O+ ion is formed
with an exceptionally strong hydrogen bond (dissociation energy about 100kJ/mol). A similar
mechanism is valid in transport of OH- ions, which also occurs along hydrogen bridges
(Figure 3).

Figure 2 Hydrogen bonding of water molecules in a tetrahedral configuration. Open circles are oxygen
atoms and closed circles are hydrogen atoms. Hydrogen bonds are represented by dashed lines [1].

Figure 3. Proton transport in water [2].


12 Jianqian Kan and Guoqing Huang

Table 2. Coordination number and distance between two water molecules [2]

Coordination number O-H…O Distance


Ice (0°C) 4.0 0.276 nm
Water (1.5°C) 4.4 0.290 nm
Water (83°C) 4.9 0.305 nm

Table 3. Comparisons of the melting and boiling points of methanol, dimethyl ether, and
water

Formula Fp/°C Kp/°C


H2O 0.0 100.0
CH3OH -98 64.7
CH3OCH3 -138 -23

1.2. Structures of Water and Ice

1.2.1. The Structure of Water (Liquid)


Due to the strong tendency of water molecules to associate through H-bridges, liquid
water is highly structured as ice, but not sufficiently established to produce long-range
rigidity. The major difference between liquid water and ice lies in the coordination number
and the distance between neighboring water molecules (Table 4).
The degree of intermolecular hydrogen bonding among water molecules is temperature
dependent. Ice at 0°C has a coordination number of 4.0, with nearest neighbors at a distance
of 2.76 Å. As the temperature increases, the coordination number increases from 4.0 in ice at
0°C, to 4.4 in water at 1.50°C, then to 4.9 at 83°C. Simultaneously, the distance between
nearest neighbors increases from 2.76 Å in ice at 0°C, to 2.9 Å in water at 1.5°C, then to 3.05
Å at 83°C.
The increase in the distance between nearest neighbors during ice-water transformation
decreases the water density, while the increase in the coordination number increases water
density. The maximum water density is observed in 3.98°C and then declines gradually. The
hydrogen-bound water structure can be changed in the presence of dissolved salts or
molecules with polar and/or hydrophobic groups. For example, in salt solutions the n-
electrons occupy the free orbitals of the cations, forming ―aqua complexes‖.
Other water molecules then coordinate through H-bridges, forming a hydration shell
around the cation and disrupting the natural structure of water. In addition, hydration shells
are also formed by polar groups through dipole-dipole interaction or H-bridges, again leading
to the disruption of the structure of water. The three-dimensional hydrogen-bound structures
of ice and water impart them with unique properties and extra energy is needed for disrupting
the structures. Table 3. lists the comparisons of the melting and boiling points between
methanol, dimethyl ether, and water.
Water 13

Figure 4. Unit cell of ordinary ice at 0°C. Circles represent oxygen atoms of water molecules. Nearest-
neighbor internuclear O-O distance is 2.76 Å; θ is 109° [3].

1.2.2. The Structure of Ice


Ice is the orderly organized crystal of water molecules. The O-O internuclear distance
between nearest neighboring water molecules in ice is 2.76 Å and the O-O-O bond angle is
about 109°, which is very close to the perfect tetrahedral angle of 109°28'. As shown in
Figure 4, each water molecule is associated with four other water molecules 1, 2, 3, and W'.
Because pure water contains H3O+, OH–, and negligible isotope variants (such as those
containing 16O, 1H, 17O, 18O, and 2H) in addition to ordinary water molecules, actual ice is not
present as the perfect crystal shown in Figure 4.
Due to the presence of H3O+, OH– and their dislocation, ice crystals suffer both
orientational and ionic defects. Only at temperatures near -180°C or lower will all hydrogen
bonds be intact, and as the temperature is raised, the mean number of intact (fixed) hydrogen
bonds will decrease gradually.
The amount and kind of solutes present in foods influence the quantity, size, structure,
location, and orientation of ice crystals. The four major ice structures are hexagonal forms,
irregular dendrites, coarse spherulites, and evanescent spherulites. The hexogonal form,
which is most highly ordered, is found exclusively in foods, provided extremely rapid
freezing is avoided and the solute is of a type and concentration that does not interfere unduly
with the mobility of water molecules.

2. STATES OF WATER IN FOODS


2.1. Water-Solute Interactions

Mixing of solutes and water results in altered properties of both water and solutes.
Hydrophilic solutes change the structure and mobility of adjacent water, and water causes
changes in the reactivity, and sometimes structure, of hydrophilic solutes. Hydrophobic
groups of added solutes interact only weakly with adjacent water. Interactions between water
and specific classes of solutes are considered below.
14 Jianqian Kan and Guoqing Huang

2.1.1. Interaction of Water with Ions and Ionic Groups


Ions and ionic groups of organic molecules hinder the mobility of water molecules to a
greater degree than do any other types of solutes. The strength of electrostatic water-ion
bonds is greater than that of water-water hydrogen bonds, but is much less than that of
covalent bonds.
The normal structure of pure water (based on a hydrogen-bonded, tetrahedral
arrangement) is disrupted by the addition of dissociable solutes. Water and simple inorganic
ions undergo dipole-ion interactions. The example in Figure 5. involves hydration of the NaCl
ion pair.
In a dilute solution of ions in water, second-layer water is believed to exist in a
structurally perturbed state because of conflicting structural influences of first-layer water and
the more distant, tetrahedrally oriented ―bulk-phase‖ water. In concentrated salt solutions,
water structure would be dominated by the ions.
The ability of a given ion to alter net structure is related closely to its polarizing power
(charge divided by radius) or simply the strength of its electric field. Ions that are small
and/or multivalent (mostly positive ions, such as Li+, Na+, H3O+, Ca2+, Ba2+, Mg2+, Al3+, F–,
and OH–) have strong electric fields and are net structure formers.
These ions strongly interact with the four to six first-layer water molecules, causing them
to be less mobile and pack more densely than HOH molecules in pure water. Ions that are
large and monovalent (most of the negatively charged ions and large positive ions, such as
K+, Rb+, Cs+, Cl–, Br–, I–, NO3– , BrO3–, IO3– and CIO4– have rather weak electric fields and
are net structure breakers, although the effect is very slight with K+. These ions disrupt the
normal structure of water and fail to impose a compensating amount of new structure.
Ions, through their varying abilities to hydrate (compete for water), alter water structure,
influence the permittivity of the aqueous medium, and govern the thickness of the electric
double layer around colloids, profoundly influence the ―degree of hospitality‖ extended to
other nonaqueous solutes and to substances suspended in the medium. Thus, conformation of
proteins and stability of colloids (salting-in, salting-out in accord with the Hofmeister or
lyotropic series) are greatly influenced by the kinds and amounts of ions present.

Figure 5. Likely arrangement of water molecules adjacent to sodium chloride. Only water molecules in
plane of paper are shown [3].
Water 15

2.1.2. Interaction between Water and Neutral Groups Possessing Hydrogen-Bonding


Capabilities
Interactions between water and nonionic, hydrophilic solutes are weaker than water-ion
interactions and about the same strength as those of water-water hydrogen bonds. Therefore,
solutes capable of hydrogen bonding might be expected to enhance or at least not disrupt the
normal structure of pure water. However, in some instances it is found that the distribution
and orientation of the solute's hydrogen-bonding sites are geometrically incompatible with
those existing in normal water. Thus, these kinds of solutes, such as urea, frequently have a
disruptive influence on the normal structure of water. It should be noted that the total number
of hydrogen bonds per mole of water may not be significantly altered by addition of a
hydrogen-bonding solute that disrupts the normal structure of water. This is possible since
disrupted water-water hydrogen bonds may be replaced by water-solute hydrogen bonds.
Hydrogen bonding of water can occur with various potentially eligible groups (e.g.,
hydroxy1, amino, carbony1, amide, imino groups, etc.). This sometimes results in ―water
bridges‖, where one water molecule interacts with two eligible hydrogen-bonding sites on one
or more solutes. A schematic depiction of water hydrogen bonding (dashed lines) to two
kinds of functional groups found in proteins is shown in Figure 8. A more elaborate example
involving a three-HOH bridge between backbone peptide units in papain is shown in Figure
9.

Figure 8. Hydrogen bonding (dotted lines) of water to two kinds of functional groups occurring in
proteins [3].

Figure 9. Examples of a three-molecule water bridge in papain; 23, 24, and 25 are water molecules [4].
16 Jianqian Kan and Guoqing Huang

2.1.3. Interaction of Water with Nonpolar Substances


The mixing of water and hydrophobic substances, such as hydrocarbons, rare gases, and
the apolar groups of fatty acids, amino acids, and proteins, enhances the hydrogen bonding of
water molecules in the vicinity of hydrophobic groups due to the repulsion with water. This
process has been termed ―hydrophobic hydration‖. Because hydrophobic hydration is
thermodynamically unfavorable, water would tend to minimize its association with apolar
entities that are present. Thus, if two separated apolar groups are present, the incompatible
aqueous environment will encourage them to associate, thereby lessening the water-apolar
interfacial area. This process is thermodynamically favorable and is referred to as
―hydrophobic interaction‖. Two aspects of the antagonistic relationship between water and
hydrophobic groups are: formation of clathrate hydrates and association of water with
hydrophobic groups in proteins. A clathrate hydrate is an ice-like inclusion compound
wherein water, the ―host‖ substance, forms a hydrogen-bonded cage-like structure that
physically entraps a small apolar molecule known as the ―guest molecule.‖ The guest
molecules of clathrate hydrates are characteristically low-molecular-weight compounds with
sizes and shapes compatible with the dimensions of host water cages comprised of 20–74
water molecules. Typical guests include low-molecular-weight hydrocarbons and halogenated
hydrocarbons; rare gases; short-chain primary, secondary, and tertiary amines; and alkyl
ammonium, sulfonium, and phosphonium salts. Interaction between water and guest is slight,
usually involving nothing more than weak van der Waals forces. Clathrate hydrates are the
extraordinary result of water's attempt to avoid contact with hydrophobic groups. There is
evidence that structures similar to crystalline clathrate hydrates may exist naturally in
biological matter, and if so, these structures would be of far greater importance than
crystalline hydrates since they would likely influence the conformation, reactivity, and
stability of molecules such as proteins.

Figure 10. Schematic depiction of a globular protein undergoing hydrophobic interaction. Open circles
are hydrophobic groups, ―L-shaped‖ entities around circles are water molecules oriented in accordance
with a hydrophobic surface, and dots represent water molecules associated with polar groups [2].
Water 17

Because exposure of protein nonpolar groups to water is thermodynamically unfavorable,


association of hydrophobic groups or ―hydrophobic interaction‖ is encouraged, and this
occurrence is depicted schematically in Figure 10. Hydrophobic interaction provides a major
driving force for protein folding, causing many hydrophobic residues to assume positions in
the protein interior. Hydrophobic interactions also are regarded as being of primary
importance in maintaining the tertiary structure of most proteins. It is therefore of
considerable importance that a reduction in temperature causes hydrophobic interactions to
become weaker and hydrogen bounds to become stronger.

2.1.4. Interaction of Water with Amphiphilic Substances


Water functions as the dispersion medium of amphiphilic compounds, such as fatty acid
salts, lipoproteins, glycolipids, polar lipids, and nucleic acids, in some foods. Water
associates with the hydrophilic entities (COO–, OH, PO4–, –C=O, or those containing the
nitrogen atom) and dissolves the compounds. Amphiphilic compounds occur as micelles in
water and each micelle contains hundreds or thousands of the molecules. The apolar groups
are directed to the interior of the micelles, while polar groups are distributed in the water
environment.

2.2. Water in Foods

Foods are composed of proteins, polysaccharides, minerals, pigments, and many other
constituents in addition to water. These constituents interact with water and significantly
affect the properties and status of water. Generally, the water in foods can be classed as ―bulk
water‖ and ―bound water‖.

2.2.1. Bound Water


―Bound water‖ is water that exists in the vicinity of solutes and other nonaqueous
constituents and binds to other solutes through covalent bonds. According to the binding
strength, bound water is further divided into the following three types:
Constitutional water: Water of this type is a constituent of other compounds and binds
the most tightly. Water in hydrates belongs to this type.
Monolayer water: Water of this type is the first layer water bound to the hydrophilic
groups of solutes. The forces involved include mainly water-ion or water-polar association,
followed by hydrogen bonding between water and solutes.
Multilayer water: Water of this type refers to water distributed in multiple layers around
nonaqueous components. The forces involved are water-water and water-solute hydrogen
bonding. Multilayer water binds tightly to nonaqueous components, but the strength is lower
than that of monolayer water. Besides, multilayer water has changed properties compared
with ordinary water.

2.2.2. Bulk Water


Bulk water or free water is not chemically bound to nonaqueous compounds and mainly
includes water that is physically entrapped. Based on the physical interaction, bulk water is
further divided into two types:
18 Jianqian Kan and Guoqing Huang

Entrapped water: Water of this type is entrapped by microstructures or ultrastructures


and cannot flow freely as pure water.
Capillary water: Water of this type is restricted in the gaps between cells or the
capillaries of food structures. Capillary water has similar reduced fluidity and vapor pressure
as entrapped water.
As mentioned above, the states of water in foods depend on the composition of foods and
the physical status of the components. Water states and contents significantly influence the
structure, processing properties, and stability of foods. The differences between bulk water
and bound water are shown in Table 4. According to Table 4, bound water and bulk water
differ in the following:

1. Bound water associates with nonaqueous constituents more tightly and its vapor
pressure is much lower than bulk water. More energy is required for removing bound
water than bulk water and the removal of bound water might irreversibly degrade the
flavor, texture, and other properties of foods.
2. Bound water freezes in much lower temperature than bulk water. This explains why
plant seeds and microbial spores can survive low temperatures. In contrast, juicy
fruits and vegetables have much higher water contents and their tissues are
susceptible to damage by ice crystals in low temperatures.
3. Bound water cannot dissolve solutes.
4. Bulk water can be utilized by microorganisms, while bound water cannot.

Table 4. Comparisons of bulk water and bound water

Item Bound water Bulk water


Occurs in vicinity of solutes and other Locates far away
nonaqueous constituents and includes from solutes and
General description
constitution water, monolayer water, occurs as water-water
and multilayer water hydrogen bonding
Not frozen even at temperatures lower Slightly lower than
Freezing point
than -40°C that of pure water
Solute solubilization
None Yes
capability
Molecular movement
compared with pure Markedly reduced or none Changed slightly
water
Enthalpy of vaporization
compared with pure Increased Nearly not changed
water
Percentage among total
water in high-moisture Less than 0.03% ca. 96%
(90%) foods

3. WATER ACTIVITY
Intensive researches have indicated that no relationship can be established between the
water content of a food with its physiochemical properties or stability. It has also been
Water 19

observed that various types of foods with the same water content differ significantly in
perishability. Thus, water content alone is not a reliable indicator of perishability. This
situation is attributable, in part, to differences in the intensity with which water associates
with nonaqueous constituents. The term ―water activity‖ (aw) was developed to account for
the intensity with which water associates with various nonaqueous constituents. Experience
shows that food stability, safety, and other properties can be predicted far more reliably from
aw than from water content.

3.1. Definition and Measurement of aw

The water activity (aw) is defined as follows:

P ERH
aw RVP
P0 100 (1)

where, RVP is the relative vapor pressure; P is the partial vapor pressure of food moisture at
temperature T; P0 is the saturation vapor pressure of pure water at temperature T, and ERH is
the equilibrium relative humidity at temperature T.
Equation (1) applies only to ideal solutions and thermodynamically equilibrium systems
and the values obtained are only approximate for food systems. The RVP of a food can be
determined by placing it in a closed chamber for a time sufficient to achieve apparent
equilibrium (constant weight) and then measuring either pressure or relative humidity in the
chamber. The vapor pressures of the aqueous solution of solutes are generally lower than that
of pure water and aw hence falls in the range 0~1.

3.2. Temperature Dependence of aw

aw is temperature dependent, and the modified Clausius-Clapeyron equation (3) can be


used to precisely present its relationship with the absolute temperature:

d ln aw H
d (1/ T ) R (3)

where, T is the absolute temperature, R is the gas constant, and ⊿H is the isosteric net heat of
sorption at the water content of the sample.
By rearrangement, equation (2-4) can be obtained:

H 1
Inaw k
R T (4)

where, R and T have the same meaning as those in Equation (2-3), ⊿H is the latent heat of
vaporization of pure water (40.5372kJ/mol), and k is calculated from the following formula:
20 Jianqian Kan and Guoqing Huang

Absolute temperature of the sample - Absolute temperature of pure water in vapor pressure p
k
Absolute temperature of pure water in vapor pressure p

Plots of Inaw versus 1/T are not always linear over broad temperature ranges, and they
generally exhibit sharp breaks with the onset of ice formation. Figure 11. is a plot of logaw
versus 1/T, illustrating that (a) the relationship is linear at subfreezing temperatures, (b) the
influence of temperature on RVP is typically far greater at subfreezing temperatures than at
above-freezing temperatures, and (c) a sharp break occurs in the plot at the freezing point of
the sample. Below freezing temperatures, the water activity (aw) can be calculated as follow:

p ff pice
aw
p0 ( SCW ) p0 ( SCW )
(5)

where, pff is the partial pressure of water in partially frozen food, p0(SCW) is the vapor pressure
of pure supercooled water, and pice is the vapor pressure of pure ice.
Two important distinctions should be noted when comparing aw values at above- and
below-freezing temperatures. First, at above-freezing temperatures, aw is a function of sample
composition and temperature, with the former factor predominating. At subfreezing
temperatures, aw becomes independent of sample composition and depends solely on
temperature; that is, in the presence of an ice phase aw values are not influenced by the kind
or ratio of solutes present. Hence, the knowledge of aw at a subfreezing temperature cannot be
used to predict aw at an above-freezing temperature. Second, as the temperature is changed
sufficiently to form or melt ice, the meaning of aw, in terms of food stability, also changes.
For example, in a product at -15°C (aw =0.86), microorganisms will not grow and chemical
reactions will occur slowly. However, at 20°C and aw 0.86, some chemical reactions will
occur rapidly and some microorganisms will grow at moderate rates.

Figure 11. Relationship between relative vapor pressure and temperature for a complex food above and
below freezing [5].
Water 21

4. MOISTURE SORPTION ISOTHERM


4.1. Definition and Zones of Moisture Sorption Isotherm

A plot of water content (expressed as mass of water per unit mass of dry material) of a
food versus aw at constant temperature is known as a moisture sorption isotherm (MSI). The
MSI of a food system is of great importance for the following reasons:

1. The ease of dehydration during concentration or drying is aw dependent;


2. The migration of water between materials during blending must be avoided;
3. It determines whether the determination of the moisture barrier properties of
packaging materials is necessary;
4. It can be used to predict the water content that inhibits microbial growth;
5. It can be used to predicate food stability.

Shown in Figure 12. is a schematic MSI for a high-moisture food plotted to include the
full range of water content from normal to dry. Omission of the high-moisture region and
expansion of the low-moisture region, as is usually done, yields an MSI that is much more
useful (Figure 13).
Several substances that have MSIs of markedly different shapes are shown in Figure 14.
Isotherms with an S shape are characteristic of most foods. Foods such as fruits, confections,
and coffee extract that contain large amounts of sugar and other small soluble molecules, and
are not rich in polymeric materials exhibit a J-type isotherm shown as curve 1 in Figure 14.

Figure 12. Schematic moisture sorption isotherm encompassing a broad range of moisture contents [3].
22 Jianqian Kan and Guoqing Huang

Figure 13. Generalized moisture sorption isotherm for the low-moisture segment of a food (20°C) [3].

Figure 14. Resorption isotherms for various foods and biological substances. Temperature 20°C, except
for number 1, which is 40°C: (1) confection (main component powdered sucrose), (2) spray-dried
chicory extract, (3) roasted Columbian coffee, (4) pig pancreas extract powder, (5) native rice starch
[6].

The MSI can be prepared in two ways. For high-moisture foods, the desorption isotherm
can be obtained by plotting the water content versus aw during dehydration. For low-moisture
foods, the resorption isotherm can be determined by plotting water content versus aw during
addition of water to the foods. The shape and position of the isotherm are determined by
several factors including sample composition, physical structure of the sample (e.g.,
crystalline or amorphous), sample pretreatments, temperature, and methodology
To deeply understanding the meaning and usefulness of sorption isotherms it is
sometimes appropriate to divide them into three zones as indicated in Figure 13.
Water 23

(1) Water present in Zone I of the isotherm is most strongly absorbed and least mobile.
This water associates with accessible polar sites by water-ion or water-dipole
interactions, is unfreezable at -40°C, has no ability to dissolve solutes, and is not
present in sufficient amount to have a plasticizing effect on the solid. It behaves
simply as part of the solid. The high-moisture end of Zone I (boundary of Zones I
and II) corresponds to the ―Brunauer-Emmett-Teller (BET) monolayer‖ moisture
content of the food. Zone I water constitutes a tiny fraction of the total water in a
high-moisture food material.
(2) Water added in Zone II occupies first-layer sites that are still available. This water
associates with neighboring water molecules and solute molecules primarily by
hydrogen bonding, is slightly less mobile than bulk water, and most of it is
unfreezable at -40°C, it exerts a significant plasticizing action on solutes, lowers their
glass transition temperatures, and causes incipient swelling of the solid matrix, leads
to acceleration in the rate of most reactions. Water in Zones I and Zone II usually
constitutes less than 5% of the water in a high-moisture food material.
(3) Further addition of water (Zone III) causes a glass-rubber transition in samples
containing glassy regions, a very large decrease in viscosity, a very large increase in
molecular mobility, and commensurate increases in the rates of many reactions. This
water is freezable, available as a solvent, and readily supports the growth of
microorganism. Zone III water is referred to as bulk-phase water. The bulk-phase
water of Zone III, either entrapped or free, usually constitutes more than 95% of the
total water in a high-moisture food.

4.2. Hysteresis of MSI

An MSI prepared by addition of water (resorption) to a dry sample will not necessarily be
superimposable on an isotherm prepared by desorption. This lack of superimposability is
referred to as ―hysteresis,‖ and a schematic example is shown in Figure 15. Typically, at any
given p/p0, the water content of the sample will be greater during desorption than during
resorption. MSIs of polymers, glasses of low molecular-weight compounds, and many foods
exhibit hysteresis. The following explanations have been proposed for the occurrence of
hysteresis:

1. Some moisture cannot be released during desorption due to the interaction with
nonaqueous components.
2. Different vapor pressures are needed for evacuating or filling the moisture entrapped
by capillaries.
3. The tissues of foods are changed during desorption. As a result, moisture cannot bind
the same tightly to the tissues during resorption and higher aw is resulted in the same
water content.
4. The magnitude of hysteresis, the shape of the curves, and the inception and
termination points of the hysteresis loop can vary considerably depending on factors
such as nature of the food, the physical changes it undergoes when water is removed
or added, temperature, the rate of desorption, and the degree of water removal during
desorption.
24 Jianqian Kan and Guoqing Huang

Table 6. Water activity and growth of microorganisms in foods [7]

Range
Inhibited Microorganisms Foods generally within this range
of aw
Highly perishable (fresh) foods and canned
Pseudomonas, Escherichia
fruits, vegetables, meat, fish, and milk;
Proteus, Shigella, Klebsiella,
1.00–0.95 cooked sausages and breads; foods
Bacillus, Clostridium
containing up to approximately 40% (w/w)
perfringens, some yeasts
sucrose or 7% sodium chloride
Salmonella, Vibrio Some cheeses (Cheddar, Swiss, Muenster,
parahaemolyticus, C. Provolone),cured meat (ham), some fruit
0.95–0.91 botulinum, Serratia, juice concentrates; foods containing up to
Lactobacillus, some molds, 55% (w/w) sucrose or 12% sodium
yeasts (Rhodotorula, Pichia) chloride
Fermented sausage (salami), sponge cakes,
Many yeasts (Candida,
dry cheeses, margarine; foods containing
0.91–0.87 Torulopsis, Hansenula,
up to 65% (w/w)sucrose (saturated) or
Micrococcus)
15% sodium chloride
Most fruit juice concentrates, sweetened
Most molds (mycotoxigenic condensed milk, chocolate syrup, maple
penicillia), Staphylococcus and fruit syrups; flour, rice, pulses
0.87–0.80
aureus, most Saccharomyces containing 15–17% moisture; fruit cake;
(bailii) spp., Debaryomyces country-style ham, fondants, high-ratio
cakes
Most halophilic bacteria, Jam, marmalade, marzipan, glacé fruits,
0.80–0.75
mycotoxigenic aspergilli some marshmallows
Xerophilic molds (Aspergillus Rolled oats containing approximately 10%
chevalieri, A. candidus, moisture; grained nougats, fudge,
0.75–0.65
Wallemia marshmallows, jelly, molasses, raw cane
sebi),Saccharomyces bisporus sugar, some dried fruits, nuts
Osmophilic yeasts
(Saccharomyces
Dried fruits containing 15–20% moisture;
0.65–0.60 rouxii), few molds
some toffees and caramels; honey
(Aspergillus echinulatus,
Monascus bisporus)
Pasta containing approximately 12%
0.50 No microbial proliferation moisture; spices containing approximately
10% moisture
Whole egg powder containing
0.40 No microbial proliferation
approximately 5% moisture
Cookies, crackers, bread crusts, etc.
0.30 No microbial proliferation
containing 3–5% moisture
Whole milk powder containing 2–3%
moisture; dried vegetables containing
0.20 No microbial proliferation approximately 5% moisture; corn flakes
containing approximately 5% moisture;
country style cookies, crackers
Water 25

Figure 15. Hysteresis of moisture sorption isotherm [5].

Figure 16. Relationships among relative water vapor pressure, food stability and sorption isotherms. (A)
Microbial growth versus p/p0. (B) Enzymatic hydrolysis versus p/p0. (C) Oxidation (nonenzymatic)
versus p/p0. (D) Maillard browning versus p/p0. (E) Miscellaneous reaction rates versus p/p0. (F) Water
content versus p/p0. All ordinates are ―relative rate‖ except for F [3].
26 Jianqian Kan and Guoqing Huang

5. WATER ACTIVITY AND FOOD STABILITY


It has been widely recognized that aw is a much better indicator of food stability than
water content. The data in Figure 16 and Table 6. provide examples of these relationships.

5.1. Water Activity (aw) and Growth of Microorganisms in Foods

Shown in Table 6. are various common microorganisms and the range of aw permitting
their growth. Most bacterial growth is affected above water activity 0.90 and most yeast and
molds, however, can grow above water activity 0.80. No microorganisms survive in water
activity lower than 0.5.

5.2. Water Activity (aw) and Chemical and Enzymatic Reactions in Foods

The relationship between water activity and the rates of chemical and enzymatic reactions
is very complex. First, water is a reactant of many chemical and enzymatic reactions and its
content significantly influences the balance of the reactions. Second, water can bind to polar
or ionic groups through hydration and significantly affects their contact with other reactants.
Third, many biomolecules swell in the presence of water and more reaction sites are exposed,
leading to accelerated reaction. However, high water contents dilute solutes and retard the
proceeding of reactions.
As shown in Figure 16, all chemical and enzymatic reactions, except oxidation reactions,
have the lowest reaction rates at the boundary of Zone I and Zone II, corresponding aw
0.2~0.3.

5.3. Water Activity (aw) and Lipid Oxidation

Figure 6.(c) indicates the relationship between lipid oxidation rate and aw. Within a
specific aw range, the rate of lipid oxidation decreases with the increase of aw. When aw
further increases to the boundary of Zone II and Zone III, the lipid oxidation rate starts to rise.
Generally, the lowest lipid oxidation rate is found in aw 0.35.
The moisture in food might either enhance or suppress lipid oxidation. When aw is lower
than 0.35, lipid oxidation is suppressed for the following reasons. Firstly, water covers the
oxidizable sites and prevents their contact with oxygen. Secondly, water hydrates ions and
eliminates the oxidation reactions initiated by ions. Thirdly, water hydrogen bonds with
hydroperoxides and retards the oxidation induced by them. Finally, water facilitates the
binding between free radicals and disrupts the chain reactions involving the free radicals.
When aw is greater than 0.35, water enhances lipid oxidation though two ways. Water
dissolves solutes and facilities their movement. Meanwhile, biomolecules swell in high water
content and more accessible sites are exposed.
Water 27

5.4. Water Activity (aw) and Maillard Reaction

Figure 16(c) illustrates the influence of aw on Maillard reaction. It could be seen that
Maillard reaction occurs mainly in aw range 0.3~0.7. In low aw, the water-solute hydrogen
bonding and the association of water with neighboring molecules retard the movement of
solutes and subsequently suppress the Maillard reaction. As the aw increases gradually,
reactants and products move more easily and the rate of Maillard reaction increase as a result.
When aw exceeds a specific value, solutes are diluted and the Maillard reaction is
consequently retarded.

5.5. Calculation of BET Monolayer Value

As shown in Figure 16, all the chemical enzymatic reactions, except oxidation, occur the
slowest in the boundary of Zone I and Zone II (corresponding to aw 0.2~0.3) and further
decrease of the water content does not change the minimum rates. The water content at the
first-encountered rate minimum is the ―BET monolayer‖ water content. The BET theory is a
theory proposed by Brunauer, Emett, and Teller in 1938 and the theory was named after them.
The BET monolayer value of a food provides a good first estimate of the water content
providing maximum stability of a dry product. One can use the BET equation to compute the
monolayer value:

aw 1 c 1
m(1 - aw ) m1c m1c (10)

where, aw is water activity, m is water content (in g H2O/g dry matter), m1 is the BET
monolayer value, and c is a constant.
From this equation, it is apparent that a plot of versus aw, known as a BET plot, should
yield a straight line. An example for native potato starch, with aw replaced by p/p0, is shown
in Figure 17. The linear relationship, as is generally acknowledged, begins to deteriorate at
p/p0 values greater than about 0.35.
The BET monolayer value can be calculated as follows:

1
Monolyaer value (m1 ) (11)
(y intercept) (slope)

From Figure 17, the y intercept is 0.6. Calculation of the slope from Figure 17 yields a
value of 10.7. Thus,

1
m1 0.088g H2O/g dry matter
0.6 10.7

In this particular instance, the BET monolayer value corresponds to a aw, of 0.2.
28 Jianqian Kan and Guoqing Huang

Figure 17. BET plot for native potato starch (resorption data, 20°C) [8].

6. FREEZING AND FOOD STABILITY


Although freezing is regarded as the best method of long-term preservation for most
kinds of foods, the benefits of this preservation technique derive primarily from low
temperature as such, not from ice formation. The formation of ice in cellular foods and food
gels has two important adverse consequences:

(1) All water converted to ice increases 9% in volume. Consequently, ice crystals formed
in a disperse system can cause locally increased pressures, which can in turn cause
mechanical damage. Hence water will leave the cells, which will shrink
considerably; enzymes will release into solution, whereby they become active; this
may result in a product of poor quality.
(2) Nonaqueous constituents become concentrated in the unfrozen phase, the unfrozen
phase changes significantly in properties such as pH, titratable acidity, ionic strength,
viscosity, freezing point (and all other colligative properties), surface and interfacial
tension, and oxidation reduction potential. In addition, solutes sometimes crystallize,
supersaturated oxygen and carbon dioxide may be expelled from solution, water
structure and water-solute interactions may be drastically altered, and
macromolecules will be forced closer together, making interactions more probable.
These changes in concentration-related properties often favor increases in reaction
rates. Thus, freezing can have two opposing effects on reaction rate: lowering
temperature, as such, will always decrease reaction rates, and freeze-concentration,
as such, will sometimes increase reaction rates.

7. MOLECULAR MOBILITY AND FOOD STABILITY


7.1. Molecular Mobility

In addition to water activity, molecular mobility (Mm) has also been used to predicate
and control the stability of foods. Molecular mobility involves all the movements of food
Water 29

components during storage that are related to the stability and processability and includes:
molecular movement or deformation caused by liquid movement or mechanical stretch;
Brownian movements or atomic rotation caused by molecular diffusion; relative movement of
materials in containers or pipelines. Some properties and behavioral characteristics of food
that are dependent on Mm are shown in Table 7. Mm is mainly influenced by hydration and
temperature. The water content and the interaction between water and nonaqueous
components determine the fluidity of the liquid phase. As temperature is increased, the
translational and rotational motion (Mm) becomes easier, while upon cooling to Tg,
translational motion of polymer segments stop.

7.2. State Diagrams

It is necessary to introduce the concept of state diagram before the discussion of the
relationship between Mm and the stability of dried, partially dried, or frozen foods, State
diagrams are supplemented phase diagrams, and contain equilibrium information as well as
information on conditions of nonequilibrium and metastable equilibrium ―states‖, and are
appropriate because foods that are dried, partially dried, or frozen do not exist in a state of
thermodynamic equilibrium. A simplified temperature-composition state diagram for a binary
system is shown in Figure 19.

Table 7. Some properties and behavioral characteristics of foods that are governed by
molecular mobility (diffusion-limited changes in products containing amorphous
regions) [9]

Dry or semidry foods Frozen foods


Moisture migration (ice crystallization,
Flow properties and stickiness
formation of in-package ice)
Lactose crystallization (―sandiness‖ in frozen
Crystallization and recrystallization
desserts)
Sugar bloom in chocolate Enzymatic activity
Structural collapse of amorphous phase
Cracking of foods during drying
during sublimation phase of freeze-drying
Texture of dry and intermediate moisture Shrinkage (partial collapse of foam-like
foods frozen desserts)
Collapses of structure during secondary
(desorption) phase of freeze-drying
Escape of volatile encapsulated in a solid,
amorphous matrix
Enzymatic activity
Maillard reaction
Gelatinization of starch
Staling of bakery products caused by
retrogradation of starch
Cracking of bakery goods during cooling
Thermal inactivation of microbial spores
30 Jianqian Kan and Guoqing Huang

Figure 19. State diagram of a binary system. Assumptions: maximal freeze concentration, no solute
crystallization, constant pressure, no time dependence. Tml is the melting point curve, TE is the eutectic
point, Tms is the solubility curve, Tg is the glass transition curve, and Tg‘is the solute-specific glass
transition temperature of a maximally freeze concentrated solution. Heavy dashed lines represent
conditions of metastable equilibrium. All other lines represent conditions of equilibrium [1].

Most foods are so complex that they cannot be accurately or easily represented on a state
diagram. Differential scanning calorimetry (DSC) can successfully determine the glass
transition temperature (Tg) of simple polymer systems, but is inapplicable to complex food
systems. The Tg of complex food systems is often determined by using dynamic mechanical
analysis (DMA) or dynamic mechanical thermal analysis (DMTA).
Glass transition temperature (Tg) is the temperature at which a supersaturated solution
(amorphous liquid) converts to a glass, and is dependent on solute type and water content. Tg‘
is a special Tg that applies only to samples containing ice, and only when ice has been formed
so maximum freeze-concentration occurs (very slow cooling). Below Tg or Tg‘ of a complex
sample, all but small molecules lose their translational mobility while retaining limited
rotational and vibrational mobility.
In the glassy state, the food will have greater stability (shelf life). As long as the
temperature remain below Tg‘, the composition of the system is virtually fixed. This implies
physical stability: crystallization, for instance, will not occur. But some chemical reactions
may still proceed, albeit very slowly because of the high viscosity and the low temperature.

7.3. Molecular Mobility, State Diagram, and Food Properties

7.3.1. Reaction Rates and Molecular Mobility


Mm is causally related to diffusion-limited properties of foods that contain, besides
water, substantial amounts of amorphous, primarily hydrophilic molecules, ranging in size
from monomers to polymers. Foods of this type include starch-containing foods, such as
pasta, boiled confections, protein-based foods, intermediate-moisture foods, and dried, frozen,
or freeze-dried foods. The utility of the Mm approach for predicting many kinds of physical
changes has been reasonably well established. However, situations do exist where the Mm
Water 31

approach is of questionable value or is clearly unsuitable. Some examples are (1) chemical
reactions whose rates are not strongly influenced by diffusion, (2) desirable or undesirable
effects achieved through the action of specific chemicals (e.g., alteration of pH or oxygen
tension), (3) situations in which sample Mm is estimated on the basis of a polymeric
component (Tg of polymer) and where Mm of small molecules that can penetrate the polymer
matrix is a primary determinant of the product attribute of interest, and (4) growth of
vegetative cells of microorganisms (p/p0 is a more reliable estimator than Mm). Examples of
diffusion-limited reactions are proton transfer reactions, radical recombination reactions,
acid-base reactions involving transport of H+ and OH-, many enzyme-catalyzed reactions,
protein folding reactions, polymer chain growth, and oxygenation/deoxygenation of
hemoglobin and myoglobin. At constant temperature and pressure, three primary factors
govern the rate at which a chemical reaction will occur: a diffusion factor, D (to sustain a
reaction, reactants must first encounter each other), a frequency-of-collision factor, A
(number of collisions per unit time following an encounter), and a chemical activation-energy
factor, Ea (once a collision occurs between properly oriented reactants the energy available
must be sufficient to cause a reaction, that is, the activation energy for the reaction must be
exceeded). For a reaction to be diffusion-limited, it is clear that factors A and Ea must not be
rate-limiting. Diffusion-limited reactions typically have low activation energies (8–25
kJ/mol). When a food is cooled and/or reduced in moisture content so that all or part of it is
converted to a glassy state, Mm is greatly reduced and diffusion-limited properties become
stable.

7.3.2. Free Volume and Molecular Mobility


The free volume of a food system decreases as the temperature decreases and the
translational and rotational motion become more difficult, which affects the motion of the
segments and local viscosity of polymers.
When the temperature decreases to below Tg, the free volume decreases significantly and
the translational motion of polymer segments stops. Hence, foods have stable diffusion-
limited properties in temperatures below Tg. The increase of free volume, which is often
unexpected, can be achieved by adding small molecular-weight solutes such as water or by
increasing the temperature. Both the two practices improve the translational motion of solutes
and are not beneficial for food stability.
However, this relationship is applicable only to certain food systems and free volume
cannot be used as a quantitative indicator of food stability to present.

7.3.3. Moisture Content and Tg


The moisture content has special effect on the Tg of food systems. The Tg of water is as
low as -135°C and water is a strong plasticizer. First, water has a much smaller size and
moves more easily than other solutes such as polysaccharides, proteins, and lipids.
The ease of motion provides space required for the movement of segments. Second,
water interacts with the polar groups on other components and replaces partial inter-molecular
or intra-molecular hydrogen bonds, which decreases the rigidity of the components and
consequently reduces the Tg. Generally, the increase of moisture content by 1% decreases the
Tg by 5~10°C.
32 Jianqian Kan and Guoqing Huang

It should be noted that the plasticizing effect is valid only when water gets entrapped in
the amorphous region of the components. In the absence of environmental effects, moisture
content is the predominant factor that affects Tg, especially in low-moisture foods.

Table 8. Relationship between the Tg and moisture content of pre-gelatinized starch and
wheat starch

Pre-gelatinized starch Native wheat starch


Moisture content Tg/°C Moisture content Tg/°C
0.153 62 0.151 90
0.166 53 0.164 67
0.181 40 0.178 59
0.222 28 0.221 40
0.247 25 0.256 33

For example, the Tg of the anhydrous mixture of 50% starch and 50% sucrose is about
60°C; when the moisture content increases to 2%, the Tg decreases to 20°C; when the
moisture content further increases to 6%, Tg falls to as low as 10°C. Table 8. lists the
relationships between the Tg and moisture content of native wheat starch and pre-gelatinized
starch. It could be seen that the Tg of both the materials increases along the decrease of
moisture content.

7.3.4. Carbohydrates, Proteins, and Tg


Carbohydrates and proteins are major constituents in many foods and their presence and
contents markedly influence the Tg of foods. Besides, the sizes of the components also affect
the Tg. Generally, carbohydrates or proteins with higher average molecular weight have more
compact structure, higher viscosity, lower free volume, and consequently higher Tg. Table 9.
lists the Tg of maltodextrin with different DE and concentration. It is seen that Tg decreases
as DE increases in the case of same moisture content. Generally, Tg is dependent on solute
type and water content, while Tg‘ is solely solute type dependent. For glycosides and polyols
with molecular weight less than 1200, Tg or Tg‘ increases as molecular weight rises. When
the average molecular weight exceeds 3000 (DE of 6 or more for starch hydrolysis products),
g becomes independent of MW, as shown in Figure 25. An exception occurs when
biomolecules are present in the entanglement networks form. In this case, g continues to
rise with increasing MW. Most biomolecules, including starch, maltodextrin, cellulose,
hemicellulose, carboxymethyl cellulose, glucan, xanthan gum, gluten, glutenin, gliadin, zein,
collagen, elastin, keratin, albumin, globulin, casein, and gelatin, have similar glass transition
curve and Tg‘, which approaches -10°C.

7.3. Mm, State Diagram and Food Stability

Knowledge on the relationship between Tg or Tg‘ and food components is of great


importance for the processing and storage of the foods.
Water 33

Table 9. Tg of maltodextrin with different Des

DE5 DE10 DE15


Moisture content Tg/°C Moisture content Tg/°C Moisture content Tg/°C
0.00 188 0.00 160 0.00 99
0.02 135 0.02 103 0.02 83
0.04 102 0.05 84 0.05 65
0.11 44 0.10 30 0.11 8
0.18 23 0.19 -6 0.20 -13

Figure 25. Relationship between average molecular weight and dextrose equivalent (DE) of commercial
starch hydrolysis products with Tg‘ [10].

7.3.1. Temperature, Mm, and Food Stability


Good correlation exists among temperature and Mm or viscosity for foods containing
amorphous regions in temperature range 10~100°C. Most molecules are present in the glassy
or rubbery state in temperatures below Tg or Tg‘. In this case, the motion of food components
is restricted and food stability is increased.

7.3.2. Food Stability Predication Based on State Diagram


The approximate stability of foods can be predicated according to the state diagram.
When foods are stored in temperatures lower than Tg or Tg‘, the diffusion of molecules is
restricted and the shelf life is markedly prolonged. In contrast, the foods are susceptible to
spoilage in temperatures higher than Tg or Tg‘. Hence, temperatures below or approaching Tg
or Tg‘ should be preferred during food storage.
Generally, Mm is more effective in predicating diffusion-limited properties, such as the
physiochemical properties of frozen foods and optimum freeze-drying conditions. However,
aw is more effective in predicating non-diffusion-limited properties and microbial growth of
34 Jianqian Kan and Guoqing Huang

ice-free foods. Because aw can be measured conveniently and quickly, it is still the major
indicator for judging food stability.

REFERENCES
[1] Fennema, OR. Food Chemistry. 3rd edition. New York: Marcel Dekker; 1996.
[2] Damodaran, S; Parkin, KL; Fennema, OR. Fennema’s Food Chemistry. 4th edition.
New York: CRC Press; 2007.
[3] Belitz, HD; Gorsch, W. Food Chemistry. 2nd edition. Berlin: Springer-Verlag, 1999.
[4] Berendsen, HJC. Specific interactions of water with biopolymers. In: Franks, F. Water -
A Comprehensive Treatise. New York: Plenum Press, 1975; 293-349.
[5] Fennema, OR. Enzyme kinetics at low temperature and reduced water activity. In:
Crowe, JH; Clegg, JS. Dry Biological Systems. New York: Academic Press, 1978; 297-
322.
[6] Ferry, JD. The evaluation of water activity in aqueous solutions from freezing point
depression. International Journal of Food Technology, 1980, 16, 21-30.
[7] Beuchat, LR. Microbial stability as affected by water activity. Cereal Foods World,
1981, 26, 345-349.
[8] Van den Berg, C. Vapour Sorption Equilibria and Other Water-Starch Interactions; A
Physico-Chemical Approach, PhD thesis, Wageningen: Wageningen Agricultural
University, 1981.
[9] Slade, L; Levine, H. Beyond water activity: Recent advances based on an alternate
approach to the assessment of food quality and safety. Critical Reviews in Food Science
and Nutrition. 1991, 30, 115–360.
[10] Levine, H., and L. Slade (1986). A polymer physico-chemical approach to the study of
commercial starch hydrolysis products (SHPs). Carbohydrate Polymers, 6, 213–244.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 3

CARBOHYDRATES

Dongfeng Wang1, Jipeng Sun2, Guoqing Huang1,3,


Xiaolin Zhou4 and Liping Sun5
1
College of Food Science and Engineering, Ocean Universityof China,
Qingdao, China
2
Third Institute of Oceanography, State Oceanic Administration,
Xiamen, China
3
College of Food Science and Engineering, Qingdao Agricultural
University, Qingdao, China
4
Department of Biology, Shantou University Medical College,
Shantou, China
5
College of Chemical Engineering, Kunming University of
Science and Technology, Kunming, China

ABSTRACT
Carbohydrates account for 3/4 of the dry weight of terrestrial plants and algae and
can be found in all the plants, animals and microorganisms that human can eat.
Carbohydrates are one of the major components in foods. The compounds not only
provide human beings with energy, but also impart foods with desired textures and tastes.
Carbohydrates undergo various changes during food processing and storage and yield
substantial compounds that affect the flavor, quality and safety of foods. This chapter
deals with the classification of carbohydrates and their most important functional
properties, in which, special attention is paid to the non-enzymatic browning reaction, its
influences on food quality, and its control. The structures, proportions, and applications
of most important polysaccharides and oligosaccharides in foods are then detailed one by
one. Dietary fiber as an important healthy diet component is also introduced at the end of
this chapter.
36 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

INTRODUCTION
Carbohydrates Classification

Carbohydrates are natural organic compounds converted from carbon dioxide and water
through photosynthesis of plants. According to the number of monosaccharide units,
carbohydrates are divided into monosaccharides, oligosaccharides and polysaccharides.
Monosaccharides are the simplest sugars in structure and can no longer be hydrolyzed. A
monosaccharide often contains three, four, five, or six carbon atoms and its functional group
might be the aldehyde or keto group. An oligosaccharide generally consists of 2 to 20
monosaccharide units and can be hydrolyzed to simple sugars. Oligosaccharides are often
found in glycoproteins or lipopolysaccharides. According to monosaccharide composition,
oligosaccharides are further divided into homo-oligosaccharides and hetero-oligosaccharides.
A homo-oligosaccharide is composed of only one type of monosaccharide, such as maltose
and dextrin with degree of polymerization less than 20. In contrast, a hetero-oligosaccharide
consists of two or more types of monosaccharide units. Polysaccharides, with degree of
polymerization greater than 20, are formed by dehydration of multiple monosaccharide units
and consist of homo-polysaccharides, such as cellulose and starch, and hetero-polysaccharide
such as seaweed and tea polysaccharides. Polysaccharides can also be classified into plant,
animal and microbial polysaccharides according to their origins or storage and functional
polysaccharides according to their biological functions.
Polysaccharides contain multiple hydroxyl groups and can covalently attach to the side
chains of proteins or peptides to form glycoproteins or protein polysaccharides.
Polysaccharides also react with carboxyl-containing molecules to form esters such as
lipopolysaccharide (LPS) and sulfate polysaccharides. Besides, polysaccharides can complex
with transition metals due to the presence of hydroxyl groups. These polysaccharide
derivatives are generally referred to as polysaccharide complexes.

Carbohydrates in Foods

Starch is one of the most abundant carbohydrates in plant-derived foods and is the most
abundant in seeds, roots and tubers. Glycogen is found in animal-derived foods especially in
muscle and liver, and it is structurally similar to amylopectin. Starch is insoluble in aqueous
solutions and does not contribute to the sweetness of foods, unless it was hydrolyzed into
oligosaccharides or glucose.
The majority of plant-derived foods contain only a small amount of free sugars and most
sugars are present as starch. For example, maize contains only 0.2%~0.5% D-glucose,
0.1%~0.4% D-fructose and 1%~2% sucrose. Free sugars not only provide the sweet taste for
foods, but also participate in flavor and color formation during thermal processing. An
increase in free sugar content during processing improves food quality. For example, to
increase the sweetness of sweet corn, sweet corn must be harvested before sugars are
converted to starch.
Many fruits are often harvested before they are fully mature for two reasons. Firstly, the
high rigidity of immature fruits facilitates their transport and storage. Secondly, starch is
Carbohydrates 37

converted to sucrose or other sweet sugars during transport and storage. This change makes
the fruits sweet and soft. The post-harvest ripening is the reverse of the starch synthesis
process in the grain, tuber and root of plants.
The contents of water-soluble sugars in processed foods are generally higher than the
corresponding materials, because sugars are intentionally added by manufacturers to meet the
requirements of consumers on flavor and color.

Carbohydrates and Food Quality

Carbohydrates are the main constitutes in foods and are closely related to the nutrition,
color, taste, texture and functionality of foods. Carbohydrates are essential nutrients for
human body and 70% of the energy needed by human body is provided by carbohydrates.
Reducing sugars with free aldehyde or ketone contribute to the formation of colors and
flavors in food thermal processing and thus affects food quality. Besides, many free sugars
are sweet and positively affect the mouth feel of foods. Some carbohydrates, such as guar
gum and carrageenan, have special viscoelastic properties and can give pleasant textures for
foods. Cellulose, pectin, and many other macromolecules can provide desired food texture
and adjust the intestine flora as dietary fibers. In addition, some polysaccharides or
oligosaccharides, such as lentinan and tea polysaccharides, have specific physiological
functions and can be directly added to foods as functional ingredients.

PHYSICOCHEMICAL PROPERTIES AND FUNCTIONS


OF CARBOHYDRATES

Carbohydrate Structure

Monosaccharide
Monosaccharide generally contains 5 or 6 carbon atoms with the general formula
Cn(H2O)n. Monosaccharides are asymmetric and optically active. Taking glyceraldehyde as an
example, the C atom in position 2 is chiral and glyceraldehyde therefore has two enantiomers.
D-glyceraldehyde is dextrorotatory and is distinguished by the prefix ―+‖ or ―d‖ and L-
glyceraldehyde is levorotatory and is labeled with prefix ―-‖ or ―l‖. Monosaccharides derived
from D-glyceraldehyde are termed D-ketones and those from L-glyceraldehyde are termed L-
ketones.
The carboxyl group of a monosaccharide ring can react with a free hydroxyl group in the
same molecule to yield more stable 5- or 6-membered hemiacetal or hemiketal, called lactol.
Lactol formation provides a new chiral center. Thus, there are two additional diastereomers
for each pyranose or furanose. These isomers are called anomers and are denoted as α- or β-
forms. Most naturally occurring simple sugars are present in the D form. Hence, it is
sometimes unnecessary to indicate the configuration of these sugars.
Monosaccharides can produce various biologically important derivatives after chemical
modification. Monosaccharide derivates that have been identified in foods include
38 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

monosaccharide phosphates, deoxy monosaccharides, glucosamines, aldonic acids, uronic


acids, saccharic acids, ascorbic acid, sugar alcohols, myo-inositol, glycosides, etc.

Sugar Alcohol, Inositol and Glycoside


Sugar alcohols are the hydrogenated products of monosaccharides and are also known as
polyols. Most sugar alcohols are the reducing products of their corresponding
monosaccharides except mannose, which occurs naturally in algae with high contents. Most
sugar alcohols are white crystals and soluble in water.

Table 3-1. Isomers of inositol


Carbohydrates 39

Sugar alcohols have lower calorific values and are sweeter than their monosaccharide
precursors. Sugar alcohols do not undergo the typical reactions of sugars and are stable
against heating and pH variations. Sugar alcohols share the same chemical properties as
common alcohols and are not involved in the Maillard reaction.
Inositol is a cyclic hexatomic alcohol and has 9 stereoisomers (Table 3-1), of which, 7 are
mesomeric and 2 are optically active. Among the isomers, only the myo isomer is biologically
active. Inositol is often present as free form in the muscle, heart, liver, and lung of animals.
The hydroxyl groups in inositol can react with phosphate acid to produce phosphoinositides.
In higher plants, all the six hydroxyl groups in inositol are phosphated to inositol
hexaphosphate. Phosphoinositides can complex with Ca2+ and Mg2+, forming calcium and
magnesium salts of phytic acid.
Glycosides are the condensation products of monosaccharides and non-saccharide ligand
through the hemiacetal hydroxyl of monosaccharide. The bond between sugar and ligand is
referred to as the glycosidic linkage.
Glycosides contain one furanose or pyranose ring and the new chiral center might be
present in the alpha or beta form. Most glycosides occur in the beta form in nature.

Oligosaccharides
Oligosaccharides are water-soluble and occur widely in nature. Generally, naturally
occurring oligosaccharides contain less than six monosaccharide units, of which, most are
disaccharides and trisaccharides. For example, sucrose and maltose are disaccharides, and
affinose is a trisaccharide.
Some high-molecular weight oligosaccharides, such as cyclodextrins (or schardinger
dextrin), have gained wide applications in the food industry. Cyclodextrins consist of 6~8 D-
glucopyranose units, corresponding to α-, β-, and γ- cyclodextrins respectively. In addition to
molecular weight, the three cyclodextrins differ in their water solubility and cavity size, as
shown in Table 3-2. X-ray diffraction reveals that α-cyclodextrin is a highly symmetric
cylinder. Six C6 hydroxyl groups are located in the bottom of the cylinder and 12 C2 and C3
hydroxyl groups are arranged on the cylinder top. The inner wall of the cylinder is covered
with C-H groups and hence more hydrophobic than the external surface. Cyclodextrins are
used to stabilize hydrophobic substances by entrapping them in the cavity during food
processing.

Table 3-2. Chemical and physical properties of cyclodextrins

Item α-cyclodextrin β-cyclodextrin γ-cyclodextrin


Number of glucose residues 6 7 8
Molecular weight 972 1135 1297
Solubility in water at 25°C (g/l) 145 18.5 232
Optical rotation +150.5 +162.5 +174.4
Inner diameter of cavity (nm) 0.57 0.78 0.95
Cavity height (nm) 0.67 0.70 0.70
40 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Polysaccharides

Structure
The degrees of polymerization (DP) of polysaccharides range from 21 to several
thousands. Polysaccharides consist of one type of structural unit (homoglycans) or multiple
types of structure units (heteroglycans) and the structural units can be linked in a linear (such
as cellulose and amylose) or branched pattern (such as amylopectin and glycogen). The
monosaccharide units generally occur periodically in polysaccharides and each periodic
repeat contains one or more alternative structural units. An example of the exceptions is the
carbohydrate components in glycoproteins, in which the monosaccharide compositions are
nonperiodic all along the chain.
The DPs of polysaccharides are heterogeneous, that is, polysaccharides have no fixed
molecular weight (MW) and display the Gauss distribution. The heterogeneity of
polysaccharide molecular weight is associated with the metabolic status of organisms. For
example, the MW of glycogen is closely dependent on the blood glucose level of animals.
When the blood glucose level is low, the liver glycogen is hydrolyzed and glycogen is
cleaved to small segments. In contrast, when the blood glucose level increases, glycogen is
synthesized in the liver and the glycogen MW is increased. In addition, many polysaccharides
are present as complexes, such as glycoproteins, glycopeptides and glycolipids. In this case,
the MWs of polysaccharides are determined by much more factors than polysaccharide alone.

Conformation
Polysaccharides are either straight or branched molecules, but they have much more
complex conformations. Some typical conformations are elucidated in the following by taking
glucans and some other polysaccharides as examples.

Extended or stretched ribbon-type conformation

This conformation is typical for 1,4-linked β-D-glucopyranosyl residues (Figure 3-1), for
instance that in cellulose fibers. This formula shows that the stretched chain conformation is
due to the zigzag geometry of monomer linkages involving oxygen bridging. The chain may
be shortened or compressed to enable formation of H-bonds between adjacent resides and
thus contribute to conformational stabilization. In this type of conformation, if the number of
monomers in turn is denoted as n and the pitch (advancement) in the axial direction per
monomer unit is denoted as h, n ranges from 2 to ±4 and h equals the length of a monomer
unit. Thus, the chain given in Figure 3-2(a) has n value of -2.55 and h value of 5.13 Å.
A strongly plated ribbon-type conformation might also occur, as shown by a segment of a
pectin chain (1,4-linked α-D-galactopyranosyl-uronate units) and an alginate chain (1,4-
linked α-L-gulopyranosyluronate units), as shown in Figure 3-3.

Figure 3-1. Conformation of 1, 4-linked β-D-glucopyranosyl residue.


Carbohydrates 41

Figure 3-2. Conformations of some β-D-glucans. Linkages: a 1→4, b 1→3, c 1→2.

(a) Peetin (b) Alginate

Figure 3-3. Plated ribbon-type conformation of pectin and alginate.

Since alginate contains multiple oxygen atoms, it can complex with many transition
metals. As shown in Figure 3-3(b), Ca2+ stabilizes the conformation of alginate. In this case,
two alginate chains are assembled in a conformation which resembles an egg box, which is
referred as the egg box type of conformation (Figure 3-4).

Hollow helix-type conformation

This conformation is typical for 1,3-linked β-D-glucopyranose units and occurs in the
polysaccharide lichenin, for example, as shown in Figure 3-5 (a). The formula shows that the
helical conformation of the chain is imposed by a U-form geometry of the monomer linkages.
Amylose (1,4-linked α-D-glucopyranosyl residues) also has such a geometry, and hence a
helical conformation (Figure 3-5 (b)).

Figure 3-4. Sketch map of egg box type of conformation.


42 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

(a) Lichenin. (b) Amylose

Figure 3-5. Conformation of lichenin and amylase.

a b c

Figure 3-6. Stabilization of helical conformations. a, clathrate compounds; b, coiled double or triple
helices; c, nesting.

The number(n) of monomers per turn and the pitch in the axial direction per residue (h)
might differ significantly in this conformation.
The value of n varies from 2 to ±10, while h can be near its limit value of 0. The
conformation of a β(1-3)-glucan, with n value of 5.64 and h value of 3.16 Å, is shown in
Figure 3-2(b).
The helical conformation can be stabilized in various ways. When the helix diameter is
large, inclusion (clathrate) compounds can be formed (Figure 3-6(a)). More extended or
stretched chains, with smaller helix diameter, can form double or triple stranded helices
(Figure 3.6(b)), while strongly-stretched chains, in order to stabilize the conformation, have a
zigzag, plated association and not stranded (Figure 3-6(c)).

Crumpled-type conformation

This conformation occurs with, for example, 1,2-linked β-D-glucopyranosyl residues


(Figure 3-2 (c)). This is due to the wrinkled geometry of the monomer O-bridge linkages.
Here, the n value varies from 4 up to −2 and h is 2–3 Å. The conformation reproduced in
Figure 3-2 (c), the n = 2.62 and h = 2.79 Å. The likelihood of such a disorderly form
associating into more orderly conformations is low. Polysaccharides of this conformational
type play only a negligible role in nature.

Loosely-jointed conformation

This is typical for glycans with 1,6-linked β-D-glucopyranosyl units, because they exhibit
a particularly great variability in conformation. The great flexibility of this glycan-type
conformation is based on the nature of the connecting bridge between the monomers. The
bridge has three free rotational bonds and, furthermore, the sugar residues are further apart.
Carbohydrates 43

Figure 3-7. Conformations of β-D-galactopyranosyl-4-sulfate and 3,7-anhydro-α-D-galactopyranosyl-2-


sulfate residues in ι-carrageenan.

Figure 3-8. Biosynthesis of ι-carrageenan.

Conformations of heteroglycans

The examples considered so far have demonstrated that a prediction is possible for a
homoglycan conformation based on the geometry of the bonds of the monomer units which
maintain the oxygen bridges. It is more difficult to predict the conformation of a heteroglycan
with a periodic sequence of several monomers, which implies different types of
conformations. Such a case is shown by ι-carrageenan, in which the β-D-galactopyranosy l-4-
sulfate units have a U-form geometry, while the 3,6-anhydro-α-Dgalactopyranosyl-2-sulfate
residues have a zigzag geometry (Figure 3-7).
Calculations have shown that conformational possibilities vary from a shortened,
compressed ribbon band type to a stretched helix type. X-ray diffraction analyses have proved
that a stretched helix exists, but as a double stranded helix in order to stabilize the
conformation.

Interchain interactions

As mentioned above, the periodically arranged monosaccharide sequence in a


polysaccharide can be interrupted by nonperiodic segments. Such sequence interferences
result in conformational disorders. This will be explained in more detail with ι-carrageenan.
44 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Initially, a periodic sequence of altering units of β-D-galactopyranose-4-sulfate (Figure 3-8, I)


and α-D-galactopyranose-2,6-disulfate (Figure 3-8, II) is built up in carrageenan biosynthesis:
When the biosynthesis of the chain is complete, an enzyme-catalyzed reaction eliminates
sulfate from most of α-D-galactopyranose-2,6-disulfate (Figure 3-7, II), transforming the unit
to 3,6-anhydro-α-D-galactopyranose-2-sulfate (Figure 3-7,III). This transformation is
associated with a change in linkage geometry. Some II-residues remain in the sequence,
acting as interference sites. While the undisturbed, ordered segment of one chain can
associate with the same segment of another chain, forming a double helix, the nonperiodic or
disordered segments cannot participate in such associations (Figure 3-9).
In this way, a gel is formed with a three dimensional network in which the solvent is
immobilized. The gel properties, e. g., its strength, are influenced by the number and
distribution of α-D-galactopyranosyl-2,6-disulfate residues, i.e. by a structural property
regulated during polysaccharide biosynthesis. The example of the ι-carrageenan gel-building
mechanism, involving a chain–chain interaction of sequence segments of orderly
conformation, interrupted by randomly-coiled segments corresponding to a disorderly chain
sequence, can be applied generally to gels of other macromolecules.
Besides a sufficient chain length, the structural prerequisite for gel-setting ability is
interruption of a periodic sequence and its orderly conformation. The interruption is achieved
by insertion into the chain of a sugar residue of a different linkage geometry (carrageenans,
alginates, pectin), by a suitable distribution of free and esterified carboxyl groups
(glycuronans) or by insertion of side chains. The inter-chain associations during gelling
(network formation), which involve segments of orderly conformation, can then occur in the
form of a double helix (Figure 3-10(a)); a multiple bundle of double helices (Figure 3-10(b));
an association between stretched ribbon-type conformations, such as an egg box model
(Figure 3-10(c)); some other similar associations (Figure 3-10(d)); or, lastly, forms consisting
of double helix and ribbon-type combinations (Figure 3-10(e)).

Figure 3-9. Schematic representation of a gel setting process.


Carbohydrates 45

Figure 3-10. Interchain aggregation between regular conformations.


a
Double helix.
b
double helix bundle.
c
egg-box.
d
ribbon–ribbon.
e
double helix, ribbon interaction.

Physiochemical Properties

Solubility
Most monosaccharides, such as sugar alcohols, glycosides, and oligosaccharides, are
water soluble. At 20°C, up to 195 g of sucrose can dissolve in 100 g of water. The solubility
of sugar alcohols varies significantly with species. For example, sorbitol has a higher
solubility than sucrose and reaches up 220 g per 100g water, while that of mannitol, erythritol
and isomaltitolto is only 17, 50, and 100 g per 100g water, respectively. Sugar alcohols
absorb much more heat than sugars when dissolution and thus produce cooling sensation in
the mouth. Sugar alcohols are added to candies and chewing gums to provide the cooling
sensation.
The solubility of glycosides is closely correlated with their ligands. Generally, glycosides
are more water soluble than corresponding ligands. For example, flavonoids are usually
insoluble, but the corresponding glycosides are soluble. Flavonoids provide foods with
different colors and tastes in the soluble glucoside form.
Each sugar unit in polysaccharides contains an average of three hydroxyl groups and each
hydroxyl group can hydrogen bond with one or more water molecules. In addition, the
oxygen atoms in the sugar ring and glycosidic bond can also form hydrogen bonds with
water. Therefore, monosaccharide units in polysaccharides can be completely solvated and
most polysaccharides have strong water-holding capabilities and are highly hydrophilic.
Polysaccharides in foods affect the movement of water and significantly influence the
functional properties of foods.
The presence of polysaccharides does not increase the penetrability or significantly
reduce the freezing point of water, although polysaccharides can be solvated by water.
Therefore, polysaccharides are good frozen stabilizers. Taking starch solution as an example,
when a starch solution is frozen, a two-phase system is formed, of which, one phase is
composed of crystal water and another phase is in the glass state consisting of 70% starch and
30% unfrozen water. Due to the extremely high polysaccharide concentration in the glass-
state phase, the viscosity is high and the movement of unfrozen water in the glass-state phase
is restricted. In addition, polysaccharides are concentrated in low temperatures. In this case,
46 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

the mobility of water is further restricted and water molecules can no longer adsorb to crystal
nucleus or the active sites for crystal growth. Both high and low molecular weight
polysaccharides can effectively protect food texture and structure from being damaged during
frozen storage.
Some polysaccharides occur in a highly ordered form. In these molecules, the chains are
tightly bound to form crystals. Because the number of exposed hydroxyl groups is
significantly reduced, these polysaccharides are insoluble in water unless the inter-chain
hydrogen bonds are broken. Taking cellulose as an example, the structural unit β-D-
glucopyranosyl residues are arranged orderly and linearly along the chain and form inter-
chain hydrogen bonds with parallel chains. The crystal regions of cellulose are not water
soluble and are very stable.
The majority of carbohydrates is not present as crystals and readily dissolves or swells in
water. Water-soluble polysaccharides and their derivates used in the food industry are often
termed gums or hydrophilic colloids. The solutions of most macromolecular polysaccharides
are viscous and the viscosity depends on the molecular size, shape, net charge and
conformation in solution. The thickening and gelling properties of polysaccharides
significantly affect the quality of foods.

Hydrolysis

Hydrolysis of Glycosides
Though glycosides occur in low contents in foods, they impart important physiological
effects and functionality to foods. For example, nature saponins are strong foaming agent and
stabilizer, flavonoids produce bitterness and color for foods. In addition to a small amount of
sweet glycosides such as stevioside and osladin, most glycosides taste bitter or astringent,
particularly when the ligand is larger than methyl. When glycosides are hydrolyzed, their
solubility is reduced and the bitterness or astringency is alleviated.
O-glycoside bonds are stable in neutral and weak alkaline solutions, but easily
hydrolyzed in acidic conditions. Most glycosides in foods (except for strong acidic foods) are
stable.
In the enzymatic hydrolysis of glycosides, the sugar component is transformed to highly
active half-chair conformation and the glycosidic bond is weakened. Then, a proton is
transferred from the enzyme to an oxygen atom in glycoside. When the oxygen atom is
separated from the carbon atom, a positively-charged carbon ion is generated. This carbon ion
then reacts with the negatively-charged -COO- group in the enzyme and is temporarily
stabilized until it is completely hydrolyzed by reacting with the -OH- group in solvent.
N-glycosidic bond is not as stable as O-glycosidic bonds and is susceptible to hydrolysis
in water. For example, glycosylamines are unstable in water and can be hydrolyzed to colored
products through a series of reactions. These reactions are the main reason of Maillard
reaction initiation (to be discussed in Section 3.2.4).
Thioglycosides, which contain S-glycosidic bonds, occur naturally in mustard and
horseradish and are very stable and water-soluble. Thioglycosides can be hydrolyzed by thio-
glucosidase to produce isothiocyanates, as shown in Figure 3-11.
Cyanogenic glycosides are another category of glycosides that significantly affect food
safety. Cyanogenic glycosides are widely present in apricot, cassava, sorghum, bamboo, and
lima beans and can yield toxic hydrocyanic acid upon hydrolysis. Amygdalin and
Carbohydrates 47

mandelonitrile are the most important cyanogenic glycosides. The complete hydrolysis of
amygdalin generates D-glucose, benzaldehyde, and hydrocyanic acid (Figure 3-12).
Table 3-3 lists the main thio-glycosides and their hydrolysates. Excessive intake of
cyanogenic glycosides leads to cyanide poisoning.
In addition to enzyme activity and environmental acidity, the hydrolysis of glycosides is
also affected by glycodic bond conformation, substitution of the sugar ring, and sugar ring
size. Generally, glycosides with β glycodic bond are hydrolyzed faster than those with α
glycodic bond. Substitution on the sugar ring reduces the hydrolysis rate and furanosides are
hydrolyzed faster than corresponding pyranosides (Table 3-4). The hydrolysis rate of
glycosides increases rapidly with elevated temperature, as shown in Table 3-4.

Hydrolysis of oligosaccharides and polysaccharides

Similar to glycosides, oligosaccharides are hydrolyzed easily by acid and enzymes and
are stable in alkaline solutions. Sucrose can be hydrolyzed by acid to yield equamolar mixture
of glucose and fructose that is called invert sugar.

Figure 3-11. Hydrolysis of thioglycosides by thio-glucosidase.

Figure 3-12. Hydrolysis of amygdalin.

Table 3-3. Main thioglycosides found in foods and their hydrolysates

Glycoside Occurrence Hydrolysates


Amygdalin and Almond kernel and dried D-Glucose + hydrocyanic acid + benzaldehyde
prunasin alpinia japonica
Linamarin Lima bean, linseed (flax), D-Glucose + hydrocyanic acid + acetone
cassava
Vicianoside Tare Vicianose + hydrocyanic acid + benzaldehyde
Linarin Sieve (black bean) and D-Glucose + hydrocyanic acid + acetone (to be
Chickpea horsebean confirmed)
48 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Table 3-3. (Continued)

Glycoside Occurrence Hydrolysates


Lotaustralin Arabicus of lotus D-Glucose + hydrocyanic acid + lotoflavin
Dhurrin Broomcorn and corn D-Glucose + hydrocyanic acid + salicylide
Sinigrin Black mustard D- Glucose + propyl isorhodanide + KHSO4
Glocoside Brassicaceae D-Glucose+ 5-ethenyl-2-oxazolidinethione, or
goitrogen+ KHSO4

Table 3-4. Effect of temperature on the hydrolysis rate of glucosidesa

Glycosides (in 0.5mol/L H2SO4) Ka


70°C 80°C 93°C
Methyl-α-D-pyran-glycoside 2.82 13.8 76.1
Methyl-β-D-fructofuranoside 6.01 15.4 141.0
Note: First order reaction rate constant, ×106 sec-1.
a

Polysaccharides are also prone to acidic or enzymatic hydrolysis, accompanying reduced


viscosity and increased sweetness. In the food industry, α-amylase and glucose glucoamylase
have been widely used in corn starch hydrolysis to produce D-glucose. The hydrolysate is
further treated by D-glucose isomerization to yield a balanced mixture of 54% D-glucose and
42% D-fructose, which is known as fructose syrup. This mixture has replaced sucrose as a
low-cost sweetener in many foods.

Oxidation
Carbohydrates with reducing free aldehydes or ketoses that can be converted to aldehyde
groups can be oxidized into aldonic acid in the presence of weak oxidants in alkaline
conditions. When strong oxidants are present, both the aldehyde and primary hydroxyl of
aldoses are oxidized to carboxyl and aldaric acids are produced as a result [3, 18].

Figure 3-13. Oxidation of D-glucose by glucose oxidase.

Some enzymes catalyze the oxidation of aldoses. For example, dehydrogenases oxidize
the primary hydroxyl of some aldoses to produce uronic acids. D-glucuronic acid, D-
galacturonic acid, and D-manuronic acid are the components of many heteropolysaccharides.
D-glucose can be oxidized to D-gluconic acid by glucose oxidase. Figure 3-13 illustrates
the preparation of D-gluconic acid and gluconolactones. D-gluconic acid-δ-lactone can be
transformed to γ-lactone and both can be hydrolyzed into D-gluconic acid at room
Carbohydrates 49

temperature. As the hydrolysis proceeds, the pH value decreases gradually. Hence,


gluconolactones can be used as a mild acidifier. Gluconolactones have gained applications in
meat, dairy and soy products and especially baked goods as a leavening agent.

Reduction
The carbonyl groups of monosaccharides can be reduced to the hydroxyl group.
Reduction of a ketose yields to two sugar alcohols isomers due to the formation of a new
chiral carbon atom. Figure 3-14 shows sugar alcohols produced by the reduction of glucose
and fructose.

Esterification and Etherification


Due to the presence of hydroxyl groups, sugars can be esterified by organic acids or some
inorganic acids, such as D-glucose-6-phosphate and D-fructose-1,6-diphosphate (Figure 3-
15). The ester derivatives of starch, such as starch succinate, have found wide applications in
the food industry. Another typical example is sucrose fatty acid ester, which is a commonly
used an emulsifier.

Figure 3-14. Reduction of D-glucose and D-fructose.

Figure 3-15. Esterification of glucose and fructose by phosphorus acid (left: D-glucose; right: D-
fructose).

Sugars can also be etherified, but naturally occurring sugar ethers are not as diverse as
sugar esters. Etherification of polysaccharides can significantly improve their functional
properties. Carboxymethyl cellulose (CMC) is an important ester of cellulose and is soluble
in water. CMC has been widely used in the food industry as thickening agent or emulsion
stabilizer.
50 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Properties of Carbohydrates

Hydroscopicity
Carbohydrates contain abundant hydrophilic hydroxyls and can bind water through
hydrogen bonding. For example, when monosaccharides or oligosaccharides are placed in
environments of different relative humidity (RH), the compounds can absorb moisture from
air (Table 3-5).

Table 3-5. Moisture adsorption of different sugars (%)

Sugar 20°C in different RHs


60%, 1h 60%, 9d 100%, 25d
D-glucose 0.07 0.07 14.5
D-fructose 0.28 0.63 73.4
Sucrose 0.04 0.03 18.4
Anhydrous maltose 0.80 7.0 18.4
Hydrous maltose 5.05 5.1 Not measured
Anhydrous lactose 0.54 1.2 1.4
Anhydrous lactose 5.05 5.1 Not measured

Figure 3-16. Hydroscopicity and water holding capacity of tea polysaccharide in different RHs (left:
RH=81%; middle: RH=43%; right: RH=43%).

All sugar alcohols, except mannitol and isomaltulose, exhibit hygroscopicity especially in
high RH environments. The hygroscopicity of sugar alcohols varies with their purity and low-
purity compounds have relatively higher hygroscopicity. Sugar alcohols are used as
moisturizing agent in cream foods and soft pastries. Polysaccharides also absorb moisture
from air and hence have good water holding capacity (Figure 3-16).
The hydroscopicity of carbohydrates, which is often referred to as moisture retention
capacity, is the most important attribute and determines their applications in foods. For
example, to overcome the sticky problem of icing sugar powders after packaging, sugars with
low hydroscopicity, such as lactose and maltose, are preferred in such products.
Carbohydrates 51

Viscosity and Gelling Property

Definition of Viscosity
Viscosity is a measure of the resistance of a fluid which is being deformed by either shear
stress or tensile stress. Viscosity can be measured with various types of viscometers and
rheometers, such as capillary viscometer, rotational viscometer, falling ball viscometer and
vibration-type viscometer.
The aqueous solutions of monosaccharides, sugar alcohols, oligosaccharides and soluble
macromolecular polysaccharides are viscous. Many factors affect the viscosity of
carbohydrates. Internal factors include the average molecular weight size and shape of
molecular chains and external factors are carbohydrates concentration, environmental
temperature, etc.

Viscosity of Polysaccharide Solutions


Polysaccharides (gums, hydrocolloids) are primarily used to thicken and/or gel aqueous
solutions and otherwise to modify and/or control the flow properties and textures of liquid
foods and beverages and the deformation properties of semisolid foods. They are generally
used in concentrations 0.25–0.50%, indicating their great ability to produce viscosity and to
form gels.
The viscosity of a polysaccharide solution is related with the molecular size, shape, net
charge and conformation of the polysaccharide in solution. Polysaccharides are often present
as random coils in solutions (Figure 3-17) and the specific conformations are closely related
to their compositions and connection modes.
Straight-chain and branched-chain polysaccharides of the same degree of polymerization
(DP) differ markedly in viscosity in aqueous solutions. Compared with their linear
counterparts of equal molecular weights and equal concentrations, solutions of branched
polysaccharides have a lower viscosity. It is assumed that the viscosity reflects the ―effective
volume‖ of the macromolecule. The ―effective volume‖ is the volume of a sphere with
diameter determined by the longest linear extension of the molecule. Molecules with larger
effective volumes undergo collision more frequently than those with smaller effective
volumes. Hence, linear polysaccharides can provide high viscosity even in low
concentrations.

Figure 3-17. Random coiled structure of polysaccharides.


52 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Figure 3-18. Effective volume of straight-chain and branched-chain polysaccharides with equal
molecular weights in solution.

Branched polysaccharides (amylopectin, glycogen) are more soluble in water than their
perfectly linear counterparts since the chain–chain interaction is less pronounced and there is
a greater extent of solvation of the molecules. This is especially the case for highly branched
polysaccharides, because they have much less effective volumes than their linear
counterparts, as shown in Figure 3-18.
In addition to the DP, unfolding degree, and rigidity, the viscosity of polysaccharides is
also affected by the shape and flexibility of polysaccharides after solvaiton.
The charge carried by solvated polysaccharides significantly influences the viscosity of
the solution. For example, straight-chain polysaccharides containing carboxyl, sulfate hemi-
ester or phosphate groups are often negatively charged. The electrostatic repulsion between
the molecules unfolds the molecules and increases their effective volume, leading to
increased viscosity. Hence, pH has significant impact on the viscosity of polysaccharide
solutions, since it is closely related to the charge statues of the molecules.
The viscosity of polysaccharide solutions decreases with the increase of temperature,
except xanthan solution, whose viscosity remains nearly unchanged in the temperature range
0~100°C. Besides, the viscosity of xanthan gum solutions decreases with shear rate increase.
Hence, xanthan gum solutions are pseudoplastic. A practical use would be in salad dressing:
xanthan gum makes it thick enough at rest in the bottle to keep the mixture fairly
homogeneous, but the shear forces generated by shaking and pouring thins it, so it can be
easily poured. When it exits the bottle, the shear forces are removed and it thickens back and
clings to the salad.

Gelation
Gelation is another important property of polysaccharides. In food processing,
polysaccharides or proteins and other macromolecules can form a sponge-like three-
dimensional network gel structure through hydrogen bonding, hydrophobic interaction, Van
der Waals attraction, ionic cross bridges, entanglement or covalent bonding (Figure 3-19).
Liquids containing small-sized solutes and polymers fill the holes of the gel network.
Carbohydrates 53

Gels possess the properties of both solids and liquids. They are not as flowable as
continuous liquids or rigid as orderly organized solids. Instead, gels are viscoelastic semi-
solids and can retain their shape even in the presence of external stresses. Polysaccharide gels
generally contain only about 1% polymer—that is, they may contain as much as 99% water.
Examples of polysaccharide gels are dessert gels, aspics, structured fruit pieces, structured
onion rings, meat-analog pet foods, and icings.
The firmness of a gel depends on the extent of junction zone formation. If the conjunction
zone area is not long enough, polysaccharide chains cannot bind tightly. In this case, the
chains can be separated by pressure or high temperature due to increased mobility of the
chains. Such gels are easily damaged and are heat-labile. If the conjunction area contains long
chain segments, the interaction between chains is strong enough to withstand applied pressure
or thermal stimulation. Gels of this type are hard and stable. Therefore, gels of different
hardness and strength are available by controlling the length of the conjunction zone.
Branched-chain polysaccharides or heteropolysaccharides cannot bind to each other well
as linear molecules and large enough conjunction areas cannot be formed between these
molecules. Hence, gels cannot be formed by branched-chain or hetero- polysaccharides. This
is also the case for charged molecules, such as those containing carboxyl groups, because the
Coulomb repulsion between chains hinders the formation of conjunction zone.

Figure 3-19. A diagrammatic representation of the type of three-dimensional network structure found in
gels.

The selection of a specific polysaccharide for a particular application depends on the


viscosity or gel strength desired, required rheology, pH of the system, temperatures during
processing, interactions with other ingredients, desired texture, and cost.

Flavor Retention
Aroma release from food matrix and the subsequent delivery of flavor to the olfactory
and gustatory receptors is greatly dependent on the type of food ingredients and on the
physicochemical properties of the aroma compound. It has been shown that macromolecules,
such as polysaccharides, are involved in the retention of volatile compounds. Polysaccharides
influence the volatility of aroma compounds and their partitioning between different phases
[1].
54 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

The use of carbohydrates may induce a significant decrease in flavor perception and/or
release. Some factors affecting the retention and release of volatile flavor compounds by
carbohydrates are depending on physicochemical properties of flavor compounds, type of
carbohydrates and their concentrations. Firstly, high molecular weight flavor compounds tend
to retain in carbohydrate than low molecular weight flavor compounds. Additionally, long
linear chain length molecules will be retained in polysaccharide matrix higher than short
chain molecules or aromatic one. Among the volatile flavor compounds such as alcohol,
aldehyde, ester and ketone, alcohol are usually the best retained in carbohydrates. The
retention of polar (hydrophilic) volatiles flavor compounds is expected to be very low in
carbohydrates complex. The second factor is depending on type of carbohydrates. Each type
of carbohydrate resents different structure that influence on the interaction between flavor
compounds and its structure and also the retention and release. Thirdly, the concentration of
carbohydrates generally shows that an increase in the concentration of sugar is proportional of
the release of flavor compounds due to the salting out effect. On the other hand, an increase in
polysaccharide concentration leads to a decrease the release of flavor compounds due to
complexation and viscosity effect of that polysaccharide themselves. This knowledge can be
used to optimize product quality in term of flavour retention during preparation or processing
and its release during eating [2].

Browning and Food Flavor


The non-enzymatic browning reaction of carbohydrates yields abundant volatile
substances in addition to melanoidin and imparts peanut, coffee bean, and bakery foods with
special flavors. Some compounds, such as maltol (3-hydroxy-2-methyl-4H-pyran-4-one) and
ethyl maltol (3-hydroxy-2-ethyl-4H-pyran-4-one), not only exhibit special flavors on their
own, but also enhance the flavors of other components in foods.
Maltol lowers the threshold of sucrose by a factor of two. Ethyl maltol is more effective
as a sweetness enhancer than maltol. Maltol enhances the sweet taste of foods, especially the
sweetness produced by sugars, and is able to mask the bitter flavor of hops and cola. Ethyl
maltol enhances the same sensation but is 4- to 6-times more powerful than maltol. In contrast
to maltol, ethyl maltol is not a natural constituent in foods.

Table 3-6. Relative sweetness of sugars and sugar alcohols to sucrose in 10% aqueous
solution

Sugar/sugar alcohol Relative sweetness Sugar/sugar alcohol Relative sweetness


Sucrose 100 D-Mannitol 69

Galactitol 41 D-Mannose 59
D-Fructose 114 Raffinose 22
D-Galactose 63 D-Rhamnose 33
D-Glucose 69 D-Sorbitol 51
Invert sugar 95 Xylitol 102
Lactose 39 D-Xylose 67
Maltose 46
Carbohydrates 55

Sweetness
Sweetness is a value relative to that of sucrose under the same conditions, which is often
referred to as 100. All sugars, sugar alcohols and oligosaccharides are sweet (Table 3-6) with
varying intensity of sweetness and some glycosides and polysaccharide complexes are
excellent sweeteners. For example, the sweetness of honey and most fruits is contributed by
sucrose, D-fructose or D-glucose. The sweetness perceived by consumers varies with the
composition, conformation, and physical aspects of sugars.
The intensities of sweetness of sugar alcohols differ markedly from those of
corresponding sugars. For example, sorbitol is sweeter than glucose and that of xylitol is
greater than xylose. Generally, all sugar alcohols, except xylitol, are less sweet than sucrose.
Sugar alcohols are usually incompletely absorbed into the blood stream from the small
intestines which generally results in a smaller change in blood glucose than sucrose. This
property makes them popular sweeteners among diabetics and people on low-carbohydrate
diets. As an exception, erythritol is actually absorbed in the small intestine and excreted
unchanged through urine, so it has no side effects at typical levels of consumption.

Nonenzymatic Browning Reactions

Nonenzymatic browning, or oxidative browning, is a chemical process that produces a


brown color in foods without the participation of enzymes. The two main forms of
nonenzymatic browning are caramelization and Maillard reaction. Ascorbic acid is also
involved in nonenzymatic browning under certain conditions. Because phenols are important
components in some foods and these compounds readily undergo autooxidation to yield
brown color, the nonenzymatic browning of phenols is also discussed in this section.

Types and Process of Nonenzymatic Browning Reactions

Maillard Reaction and Its Process


The Maillard reaction is a form of nonenzymatic browning and involves the complex
reactions between reducing sugars and amino acids/proteins. French chemist Louis-Camille
Maillard described the reactions for the first time in 1912.
John Hodge then named the reaction after Maillard and summarized the reaction process
for the first time in 1953, as shown in Figure 3-20. The process of the Maillard reaction is
often divided into three stages.
The initial stage of the Maillard reaction involves the condensation of a carbonyl group,
for example from a reducing sugar such as glucose, with a free amino group, typically the
epsilon amino group of lysine residues within proteins. This glycation reaction results in the
formation of an unstable Schiff base (aldimine) that spontaneously rearranges to the more
stable 1-amino-1-deoxy-2-ketose (ketoamine), which is also known as the Amadori product
after the Italian scientist Mario Amadori. When the initial sugar is glucose, the Amadori
product is commonly known as fructoselysine (FL) (Figure 3-21).
56 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Figure 3-20. Maillard reaction scheme recommended by John Hodge [3].

Amadori products are degraded via various pathways in the intermediate stage, leading to
the formation of furfurals, reductones and fragmentation products (carbonyl and
hydroxycarbonyl compounds). Furfural or hydromethylfurfural (HMF) formation (Figure 3-
22) is favored under acidic conditions, while alkaline media favor the production of
reductones (Figure 3-23).
The products of 1-amino-1-deoxy-2-ketose fragmentation in alkaline media, such acetol,
diacetyl, pyruvaldehyde, etc, are able to react with amino acids via the Strecker degradation
(named after the German chemist Adolph Strecker, Figure 3-24) to give Strecker aldehydes of
the amino acids and aminoketones; the latter subsequently condense to form pyrazines.
Strecker aldehydes and pyrazines contribute to aroma formation in heated foods.
Carbohydrates 57

Figure 3-21. Scheme of the initial stage of the Maillard reaction.

Figure 3-22. Formation of hydromethylfurfural from 1-amino-1-deoxy-D-fructose.

Figure 3-23. Formation of reductones from 1-amino-1-deoxy-D-fructose.

Figure 3-24. Scheme of the Strecker degradation.


58 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Figure 3-25. Scheme of the aldolisation of aldehydes and ketones.

The majority of the compounds displaying color are formed in the final stage of the
Maillard reaction. Furfurals, reductones, aldehydes either undergo aldol condensation without
the intervention of amino compounds or react with amino compounds as other intermediates,
leading to the ultimate reaction products known as melanoidins. Hodge defined melanoidins
as ‗brown, nitrogenous polymers and copolymers‘. The structures of melanoidins remain
unknown till now. The pigment generated in the Maillard reaction is soluble in the early stage
of polymerization and exhibits no characteristic absorption pick in the visible range. Infrared
spectrum and chemical composition analysis indicate that melanoidins contain unsaturated
bonds, heterocycles, and complete amino acid residues.
The reaction mechanism of the Maillard reaction proposed by John Hodge is largely
unchanged after 60 years and is still widely cited. However, the hypothesis has some flaws.
Firstly, the mechanism presents only the general process of the Maillard reaction and the
details on the reaction are not described. Secondly, some new reactions have been identified
by other researchers. For example, the work of Japanese scientists Namiki and co-workers
found that the carbonyl fission products can also be formed directly from N-substituted
glycosylamine through a free radical-mediated pathway, which is known as the Namiki
pathway [4].

Mechanism of Caramelization
Heating of carbohydrates without the presence of nitrogen-containing compounds causes
a complex group of reactions termed caramelization. Unlike the Maillard reaction,
caramelization of carbohydrates is thermolysis as compared to the reaction with amino acids.
Mild heating or thermolysis in the early stage leads to anomeric shifts, ring size change,
glycosidic bond breakdown, and new glycosidic bond formation. Mostly, thermolysis causes
dehydration of the sugar molecule with introduction of double bonds or formation of anhydro
rings. Caramelization yields hundreds of flavor compounds and these compounds impart
foods with pleasant color and flavor. The caramels derived from different sugars have similar
compositions. Generally, the products of caramelization include caramel, which is the
polymerization product of sugar dehydration, and aldehydes, ketones, phenols, etc.
The caramelization process is divided into two stages.

Caramel formation

Figure 3-26 illustrates the formation of caramel by taking sucrose as the example. It can
be seen that caramelization is the removal of water from a sugar, proceeding to isomerization
and polymerization of the sugars into various high-molecular weight compounds.
Carbohydrates 59

The aqueous solutions of caramels are colloidal and the isoelectric points range from
3.0~9.0, with some exceptions with pI lower than 3. The presence of acids or certain salts
facilitates the reaction. The caramel prepared by heating sucrose solution in the presence of
acid or acidic ammonium salts has been widely used in food coloration.
The caramelization of sucrose consists of three steps:
Step 1: Caramelization of sucrose starts with the melting of the sugar at high
temperatures followed by foaming (boiling). At this stage sucrose decomposes into glucose
and fructose. This is followed by a condensation step, in which the individual sugars lose
water and react with each other to form the isosaccharosan (Figure 3-27). Isosaccharosan
loses the sweetness of sugar and tastes bitter instead.

Figure 3-26. General mechanism for thermal promoted caramelization of sucrose.

Figure 3-27. Structure of isosaccharosan.

Step 2: This step involves further dehydration reactions. Isosaccharosan dehydrates and
condensates to caramelan. Caramelan is one of the three main products of sucrose
caramelization. Caramelan are lightly brown and tastes bitter with formula of C24H36O18.
Caramelan melts at 138°C and is soluble in water and ethanol.

Step 3: This step includes both fragmentation reactions (flavor production) and
polymerization reactions (color production). With respect to color production, caramelan is
further dehydrated to form caramelen. If the mixture is further heated, polymerization
reaction occurs and the insoluble caramelin is generated. The caramelan melts at 154°C,
tastes bitter, and is soluble in water with formula of C36H50O25. Caramelin appears dark
brown and are not soluble in water with formula of C125H188O80.
60 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

The presence of iron enhances the color of caramels. Phosphates, inorganic salts,
alkalines, citric acid, ammonia, and ammonium sulfate catalyze sugar caramelization.
Ammonia and ammonium sulfate increase the yield of caramel. However, the addition of
ammonia and ammonium sulfate cause the generation of 4-methylimidazole at high
temperatures, which is an anticonvulsants and can cause nervous system damage after long-
term consumption. Ammonia and ammonium sulfate have been prohibited in the production
of caramels.

Caramel flavors from thermal fragmentation

a. Formation of aldehydes in acid media.


In acid media, aldoses or ketones undergo enolization to yield hexose-1,2-enediol when
heated, followed by a series of dehydration reactions, as shown in Figure 3-28.
b. Formation of aldehydes in alkaline media.
In alkaline conditions, the intermediate 1,2-enol hexose, such as fructose, is produced via
the tautomery of reducing sugars and is then fragmented when heated, as shown in Figure 3-
29.

Figure 3-28. Formation of aldehydes from sugars in acid media and thermal condition.
Carbohydrates 61

Figure 3-29. Thermal fragmentation of 1, 2-enediol-hexose in alkaline media.

Mechanism of Nonenzymatic Browning of Ascorbic Acid


Ascorbic acid is a well-known natural antioxidant. It can be readily oxidized in two ways.
In the presence of oxygen, ascorbic acid is oxidized to dehydroascorbic acid, which is then
dehydrated to yield 2,3-diketogulomic acid (DKG). DKG is further decarboxylated to yield
xylosone and subsequently reductones, which participate in the reactions in the intermediate
and final stages of the Maillard reaction. Ascorbic acid degrades very quickly in the presence
of oxygen and the specific rate is related to the level of dissolved oxygen and extraneous gas.
If the food matrix contains components with reduction potential higher than that of
ascorbic acid, ascorbic can be oxidized to dehydroascorbic acid even in the absence of
oxygen. In both cases, dehydroascorbic acid is then transformed to DKG in the presence of
water. DKG further undergoes decarboxylation and dehydration to yield aldofuranoses or
reductions. These products can participate in the Maillard reaction, leading to the formation
of brown pigments (Figure 3-21). Ascorbic acid is slowly oxidized to dehydroascorbic acid in
acidic solutions with pH < 5 and this process is reversible.

Mechanism of Polyphenols Browning


Some plant-derived foods contain high levels of phenolic compounds. For example, the
content of polyphenols in green tea is up to 30%. The phenolic hydroxyl group of
polyphenols is very susceptible to oxidation, especially in alkaline conditions. Polyphenols
undergo auto-oxidation at high temperature and high moisture conditions and the oxidation
products exhibit different colors. Polyphenols are an important cause of food browning and
the browning mechanism is elucidated by taking catechin as an example.
62 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Figure 3-30. Mechanism of ascorbic acid browning.

Structure of catechin

Catechin possesses two benzene rings (called the A- and B-rings) and a dihydropyran
heterocycle (the C-ring) with a hydroxyl group on carbon 3, as shown below:

Catechin contains multiple phenolic hydroxyl groups and is very susceptible to oxidation,
polymerization, and condensation. Catechin is white crystal and is oxidized to yellowish-
brown compounds in the air. Catechin is soluble in water, ethanol, methanol, acetone, and
acetic anhydride, partially soluble in ethyl acetate and acetic acid, and insoluble in chloroform
and anhydrous ether.
Carbohydrates 63

Mechanism of oxidation

The hydroxyl groups in catechin have different activities with respect to auto-oxidation.
Hydroxyl groups in adjacent and vicinal positions are oxidized easily, while the –OH in
carbon 3 on ring C cannot be oxidized.
The mechanism of the nonenzymatic auto-oxidation of catechin is very complex and not
well understood to the present. It is generally accepted that the process involves two
reactions. The first is the formation of quinine. Quinine is unstable and rapidly undergoes
condensation. The condensation product in the early stage is soluble, light yellow and tastes
bitter. As the reaction proceeds, the intermediates are further condensed to brown nitrogenous
polymer in the presence of amino compounds. The general process is illustrated in Figure 3-
31.

Effects of Nonenzymatic Browning on Foods


Nonenzymatic browning reaction is one of the most important reactions during the
storage and processing of foods. A large number of food components, such as carbohydrates,
amino acids, phenolic compounds, and ascorbic acid, are involved in the reaction. The
reaction yields multiple volatiles and nonvolatile compounds, which significantly influence
the color, aroma, taste, nutrition, and safety of foods.

Figure 3-31. Mechanism of nonenzymatic browning of catechin.

Effects on Color
The composition of nonenzymatic browning reaction products is very complex. Many
components affect the color of foods and their molecular weights range from several
hundreds to up to more than 100000 Dolton. Several coloring compounds have been isolated
from different model systems.
64 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Xylose – Lys model system

A yellow compound is isolated from the Maillard reaction product of the xylose – Lys
model system and MS and NMR reveals that its possible structure is one of the following two
[5]:

(1)

Furan-2-carboxaldehyde – L-alanine model system

Two red products are isolated and identified from the Maillard reaction product and their
structures are shown below [6]:

(2)

Xylose – L-alanie model system

A white compound is isolated:

(3)

Glucose – propylamine model system in ethanol solution.

A yellow product is identified:

(4)

Xylose – alanie model system in the presence of carbonyl compounds.

Two yellow compounds are isolated:


Carbohydrates 65

(5)

One red compound is isolated:

(6)

Starting materials, as well as reaction conditions, markedly affect the elemental


composition and structure of melanoidins. Thus far, the structures of the melanoidins have not
been elucidated, although some structural insights have been gained from model reactions. In
a glucose/amino acid Maillard reaction system, the carbonyl compound reacts mainly via the
Amadori product to form several deoxyosones which are able to react with each other in an
aldol-type condensation to form a basic melanoidin skeleton of amino-branched sugar
degradation products. Figure 3-32 shows the possible structure of a melanoidin formed from
3-deoxyhexosuloses in this way.

Figure 3-32. Part of possible melanoidin structure formed from 3-deoxyhexosulose involving amino
compounds [7].

Figure 3-33. Strecker degradation of L-Lys with dicarbonyl compound.


66 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Figure 3-34. pH changes of various sugar-amino acids solutions heated at 100°C. From up to down:
glucose, lactose, lactose + alanine, glucose +alanine, lactose + glycine, glucose + glycine, glucose +
lysine, lactose+ lysine.

Effects on Food Aroma and Taste


The intermediate and final products of nonenzymatic browning reactions markedly
influence the aroma and taste of foods. Under high temperatures, carbohydrates undergo
dehydration, fragmentation, isomerization, and oxidation/reduction, giving rise to formic acid,
acetic acid, lactic acid, acetol, acetoin, diacetyl, pyruvic acid, and many other products. The
dicarbonyl compounds generated in non-enzymatic browning can initiate the changes of other
food components. For example, amino acids are deaminated and decarbonylated to yield
abundant aldehydes, as shown in Figure 3-33:
Non-enzymatic browning can produce both desirable and undesirable flavors. Maltol and
isomaltol in breads gives the caramel-like flavor for breads and 4-hydroxy-5-methyl-3(2H)-
furanone delivers the flavor of roasted meat and is used as flavor and sweetness enhancer,
while some pyrazine and aldehyde compounds are the source of undesired burned taste in
some foods.
CO2 is a product of the Strecker degradation and its volume released is proportional to
the moiety of dicarbonyl compounds. Besides, the reducing ketones and aldehydes produced
can be readily oxidized to acidic compounds. Therefore, non-enzymatic browning reaction
reduces the acidity of foods. Figure 3-34 shows the changes of pH in different model systems.

Antioxidation
The antioxidant activity of Maillard reaction products (MRPs) was firstly reported by
Franzke and Iwainsky in 1954. The two researchers found that the thermal reaction product of
glycine and glucose improved the stability of margarine against oxidation [8]. However, this
research did not attract much attention until 1980s. The antioxidant components in various
MRPs are not identified yet and the antioxidant activity of MRPs is studied mainly by using
various model systems.
Carbohydrates 67

Elizalde et al. found that the volatile compounds of the MRP of the glucose-glycine
system showed a significant antioxidant activity and prolonged the induction period of
soybean oil thermoxidation by up to 3 times in relation to the control [9]. Bedinghaus and
Ockerman compared the lipid oxidation inhibitor activities of fifteen MRPs prepared by
heating one of three sugars (glucose, xylose, and dihydroxyacetone) with one of five amino
acids (arginine, histidine, leucine, lysine, and tryptophan) by using cooked ground pork
patties as the model food. It was found that the most effective MRPs were xylose-lysine,
xylose-tryptophan, dihydroxyacetone-histidine, and dihydroxy-acetone-tryptophan when
compared to controls [10]. Yamaguchi et al. obtained a fraction from the MRP of the xylose –
glycine system through multiple chromatographic steps and found that it is more effective in
preventing linoleic acid oxidation than BHA and propyl gallate [11]. Yoshimura et al.
investigated the MRP of the glucose-glycine system on the inhibition toward active oxygen
and fount that it inhibited more than ca. 90% of active oxygen species existing in the form of
hydroxyl radicals (•OH) [12].
Morales and Jiménez-Pérez investigated the DPPH· scavenging activities of MRPs
produced by heating glucose or lactose with lysine, alanine or glycine. It was found that all
the MRPs were effective DPPH· scavenging agents. Browning was not directly related to the
free radical scavenging properties of MRPs formed at prolonged heating conditions and
fluorescence measurement is more effective than browning to follow the formation of MRPs
with free radical scavenging activities, as shown in Figure 3-35.
Although the antioxidant effects of MRPs have been well recognized, their application as
effective antioxidants in foods is still limited due to insufficient knowledge on the structure
and antioxidant mechanism of MRPs. Early researches indicated that antioxidant capability of
MRPs is contributed by the intermediate reductones of the Maillard reaction and the metal
chelating ability of MRPs. Latest researches indicate that MRPs also have strong active
oxygen scavenging capabilities and can reduce peroxides.

Note: A: up to down: glucose+alanine, glucose+glycine, glucose+lysine.


B: up to down: lactose+alanine, lactose+glycine, lactose+lysine.

Figure 3-35. Free radical scavenging activity in sugar/amino acid mixtures heated at 100°C up to 24 h
[13].
68 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Effects on Nutrition
One of the most obvious negative consequences of the Maillard reaction in food is the
loss of nutritive value of proteins due to decreased digestibility, destruction and/or biological
inactivation of amino acids, including essential amino acids like lysine and tryptophan,
inhibition of proteolytic and glycolitic enzymes, and interaction with metal ions.
Lysine is the most liable to loss in non-enzymatic browning, followed by alkaline amino
acids, including L-Arg and L-His. In addition to sugar-amino acid reactions, Strecker
degradation is also involved in amino acid loss. The formation of complex macromolecules
reduces the solubility of proteins and reduces their nutritional value. The effect of the
Maillard reaction on amino acid availability has been investigated by using rainbow trout
(Salmo gairdneri) as the model. It was found that arginine and lysine exhibited the greatest
losses in the mixture of fish protein isolate and glucose stored for 40 d at 37°C and the
apparent digestibility and absorption of individual amino acids, particularly lysine, was lower
in trout fed browned protein than in those fed the control protein [14].
Vitamin C is also involved in browning and therefore suffers loss during the process.
The products of non-enzymatic browning reduce the bioavailability of minerals.
Whitelaw et al. employed the dialysis procedure to determine the effect of the Maillard
reaction on apparent 65Zn availability. In the presence of 65ZnCl2, the amino acids glycine, D-
leucine, L-proline, L-lysine and L-glutamic acid were combined with D-glucose and
autoclaved (110°-120°C, 15 atm, 10 min) to produce high molecular weight 65Zn binding
compounds that were not dialyzable (6-8KD). Experiments in stimulated gastrointestinal
digestive conditions revealed that the Maillard reactions significantly reduced the
bioavailability of Zn [15].

Harmful Compounds
The safety of MRPs has attracted wide attentions in recent years. With the development
of new instrumental analysis measures, more and more harmful compounds have been
isolated and identified. For example, mutagenic compounds have been found in instant and
caffeine-free coffee and the compounds consist of dicarbonyl compounds, methylglyoxal,
diacetyl and glyoxal, among which the methylglyoxal presented highest mutagenic activity
[16]. Also in both fried and grilled meat and fish, mutagenic compounds were identified,
mainly stemming from heterocyclic amines [17]. The reaction mechanism seems to have a
major influence on the mutagenicity of the reaction products. For instance, ketose sugars
showed a higher mutagenic activity than the corresponding aldose sugars [18]. However, due
to the complexity of non-enzymatic browning reactions and the poor stability of
intermediates, only very few harmful compounds have been well elucidated, of which,
acrylamide is the most intensively studied.
Acrylamide is a well-known cancerogen and can cause neurologic damage. Acrylamide
has been detected in nearly all the foods, but fried and roasted foods that are processed at high
temperatures have much higher acrylamide contents, as presented in Table 3-7.
Carbohydrates 69

Table 3-7. Content of acrylamide in some foods

Foods Acrylamide contents (μg/kg) Number of


Median Minimum ~ maximum samples
Potato chips 1,200 330-2,300 14
French fries 450 300-1,100 9
Biscuit 410 <30-650 14
Fried bread 140 <30-1,900 21
American breakfast 160 <30-1,400 15
Cornflakes 150 120-180 3
Bread 50 <30-160 20

Factors Influencing Nonenzymatic Browning

Type of Sugar and Amino Acid


The structures of sugars and amino acids significantly affect the rates of the non-
enzymatic browning reactions between them. Among common reducing sugars, pentoses are
10 times more susceptible to non-enzymatic browning than hexoses. Under the same
conditions, the reactivity of some pentoses follow the sequence ribose > arabinose > xylose
and that of some hexoses are galactose > mannose > glucose.
Among carbonyl compounds, -hexenal has the highest browning rate, followed by -
dicarbonyl compounds and ketones in sequence.
Alkaline amino acids undergo browning more readily than acidic and neutral amino acids
and those with the amino group locating at the end or ε- position are more susceptible to
browning than those with amino group at the α- position.
Non-enzymatic browning also occurs between proteins and carbonyl compounds, but the
reaction is slower than that of peptides.

Temperature and Time


Reaction temperature significantly affects browning. When the temperature increases by
10 °C, the browning rate can be 3~5 times higher. Generally, non-enzymatic browning
occurs rapidly at temperatures higher than 30 °C.
The degree of browning is also affected by reaction time. When the mixture of
disaccharide or monosaccharide and amino acid is heated at 100°C, the absorbance of the
solutions at 420nm increases with heating time, indicating that the formation of the brown
color is positively correlative with the reaction time (Figure 3-36).
In addition to the color and flavor of foods, reaction temperature and time influence the
generation of harmful compounds as well. For example, when equimolar amount of
asparagine and glucose is heated, the formation of acrylamide is detected in 120 °C and the
content continues increasing with temperature elevation until 170 °C. The formation of
acrylamide generated in the mixtures of glucose and glutamine, methionine, or asparagine
heated at 180 °C as a function of time (5~60 min) reveals that the contents of acrylamide
varies with amino acids. The highest acrylamide content is found in the glucose/asparagine
system and the highest content appears in the 5th minute. When the reaction proceeds, the
level of acrylamide keeps decreasing. In the glucose/glutamine system, the highest
70 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

acrylamide formation is detected within 10 min and the content remains unchanged thereafter.
While in the glucose/methionine system, the formation of acrylamide keeps increasing in the
first 30 min and then remains unchanged in remaining time.

pH
Medium pH affects multiple reactions of non-enzymatic browning. Aqueous solutions of
sugar (xylose or glucose) and amino acid (glycine or lysine monohydrochloride), are heated
without pH control for up to 120 min and the total reaction products are analyzed by HPLC
and compared with those of the corresponding model systems maintained at pH 5 throughout
heating.
For xylose-lysine, seven of these peaks are common to the systems heated both with and
without pH control for 15 min, while no peak is common to both glucose-lysine systems
heated for 120min, indicating different products are formed under the conditions [19].
Generally, the browning rate between sugars and amino acids increases with pH when the
medium pH is greater than 3.5 and is reversely proportional to pH values in the range 2.0~3.5.
Hence, browning can be suppressed by reducing pH. This is why browning does not occur
readily in acidic foods such as pickled vegetables. Sulfites inhibit nonenzymatic browning by
reacting with carbonyl intermediates, thereby preventing their further reaction to form brown
pigments [20].

Water Content and Metals


The rate of non-enzymatic browning is a function a water activity. Generally, non-
enzymatic browning occurs readily in the water activity range 10%~15% and is suppressed in
water activity less than 3%. Moderate water content facilitates the mobility of solutes, but too
high water contents lead to solute dilution and consequently reduced browning.
Metal ions are also involved in the process of non-enzymatic browning, as evidenced by
the fact that the browning of grapefruit juice is inhibited by the addition of a chelating agent
EDTA [21]. Cu(I), Cu(II), Fe(II), and Fe(III) speed up the browning of ascorbic acid and
phenols, but other metals, such as Pb, Zn, and Sn, seem to have little effect.

Figure 3-36. The time course of the browning develop in heated sugars -amines solutions at 100°CUp
to down: glucose+lysine, lactose +lysine, glucose+glycine, lactose+ glycine, glucose+alanine, lactose
+alanine, lactose, and glucose.
Carbohydrates 71

High Pressure
High pressure (100-1000 MPa), which is gaining increasing importance as a food-
processing technology particularly in combination with moderate temperatures (30-60 °C),
may influence the Maillard reaction and thereby affects the flavor, color, and nutritional value
of foods. It has been proposed that high pressure exerts its influence on non-enzymatic
browning by changing the medium pH. The effect of high pressure on Maillard reaction has
been investigated by using the glucose-lysine model system over a range of pH values (5-10)
at 60 °C either under atmospheric pressure or at 400 MPa. The results obtained showed that
high pressure affected in different ways the different stages of the Maillard reaction and that
such effects were strongly influenced by pressure-induced changes in the pH of the systems.
In unbuffered media, at an initial pH 8.0, the formation of Amadori rearrangement products
(ARP) was not considerably affected by pressure, whereas the intermediate and advanced
stages of the Maillard reaction were suppressed, suggesting a retardation of the degradation of
the ARP. In buffered media, at pH values 8.0, pressure slowed the Maillard reaction from the
initial stages. These effects are attributed to the pH drop caused by the pressure-induced
dissociation of the acid groups [22].

Control of Non-Enzymatic Browning


The non-enzymatic browning has both beneficial and adverse impacts on food qualities.
Researchers have developed rational approaches to minimize adverse consequences of
browning reactions and optimize beneficial ones.
Non-enzymatic browning can be prevented by chemical or biochemical methods [23]:

Sulfhydryl compounds

Sulfur-containing amino acids such as cysteine, N-acetylcysteine, and the tripeptide


glutathione play key roles in the biotransformation of toxic compounds by actively
participating in their detoxification. These antioxidant and antitoxic effects are due to a
multiplicity of mechanisms including their ability to act as (a) reducing agents, (b) scavengers
of reactive oxygen (free radical species), (c) destroyers of fatty acid hydroperoxides, (d)
strong nucleophiles that can trap electrophilic compounds and intermediates, (e) precursors
for intracellular reduced glutathione, and (f) inducers of cellular detoxification. Under certain
conditions, SH-containing compounds may be as effective as sodium sulfite in preventing
nonenzymatic browning of both apples and potatoes.

Acetylation of amino groups

Modifications of amino groups prevent them from participating in browning reactions.


For example, treatment of foods with the enzyme transglutaminase will transform lysine
amino groups to amide groups. The former initiate browning, whereas the latter do not.

Antioxidants

Oxygen seems to be required for some nonenzymatic browning reactions. Hence,


antioxidants could suppress browning in some foods. Besides, antioxidants can trap or
72 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

prevent the formation of intermediates in the Maillard and related reactions, thus prevent the
formation of undesirable compounds during food processing.

Deglycation

An extract from soil microorganisms catalyzed the deglycation of α- and ε-


fructosyllysines to lysine. This finding suggests that these purified enzymes could be used to
prevent or reverse Maillard reactions in foods and in vivo provided they are safe in other
regards.
Non-enzymatic browning can also be prevented by controlling processing conditions:

Low temperature

Non-enzymatic browning occurs slowly in low temperatures. Storage at low temperatures


can delay non-enzymatic browning in foods.

Sulfurous acid

The condensation product of carbonyl group and sulfurous acid can react with R-NH2
and the resultant product cannot be transformed to Schiff‘s base. Hence, SO2 and sulfites
suppress non-enzymatic browning.

pH

Non-enzymatic browning does not occur readily in acidic conditions as in alkaline


conditions.

Product concentration

Products with lower concentrations suffer lighter browning. For example, because lemon
juice is more susceptible to browning than orange juice, the concentration factor of lemon
juice is 4:1, which is lower than 6:1 of orange juice.

Insensitive sugar

The presence of free carbonyl groups is essential for non-enzymatic browning.


Replacement of reducing sugar with sucrose can prevent browning.

Removal of sugar

Some foods contain only trace amount of sugars and these sugars can be removed to
prevent non-enzymatic browning. For example, glucose oxidase and catalase have been used
to remove the trace glucose in dried egg yolk and dehydrated meat.

Calcium salts
Calcium can complex with amino acids to form precipitates.
Carbohydrates 73

IMPORTANT OLIGOSACCHARIDES AND POLYSACCHARIDE IN FOODS


Oligosaccharides and polysaccharides occur widely in the nature and some of them
contribute largely to the quality and nutrition of foods. Oligosaccharides and polysaccharides
have gained important applications in the food industry as thickening agent, gelling agent,
crystallization inhibitors, clarifying agent, stabilizing agent, film forming agent, flocculating
agent, controlled-release agent, expansive agent and encapsulation agent. Some carbohydrates
posses important physiological functions and many functional foods have been developed
based on them.

Oligosaccharides

Oligosaccharides occur naturally in a variety of foods, especially plant-derived foods,


such as vegetables, grains, legumes, and algae. Oligosaccharides have also been identified in
animal-derived foods, including milk, honey and insects. Sucrose, maltose, lactose and
cyclodextrin are the most important oligosaccharides for the food industry. Some
oligosaccharides, such as fructo-oligosaccharides, xylo-oligosaccharides, and Konjac
oligosaccharides, have obviouse physiological functions and can be used as the effective
ingredients of functional foods.
Cellobiose, maltose, isomaltose, gentiobiose and trehalose are common disaccharides. All
these disaccharides, except trehalose, contain a free semi-acetal group and hence are reducing
sugars.
Sucrose, lactose, lactulose, and melibiose are hybrid-oligosaccharides and each contains a
reducing group except sucrose. Of the sugars, lactose deserves special attention. Lactose is
found notably in milk and non-fermented dairy products and accounts for 2~8% of milk by
weight. In small intestine, lactose is hydrolyzed by lactase to D-glucose and D-galactose,
which are then absorbed by the human body. However, some consumers lack the lactase and
cannot digest or metabolize lactose. These consumers might suffer abdominal pain, bloating,
flatulence, diarrhea, nausea, and acid reflux and this is called lactose intolerance or lactase
deficiency.
Some natural foods contain functional oligosaccharides. These compounds cannot be
absorbed by human body and provide very few energy, but can enhance the proliferation of
intestinal bifidobacteria and prevent dental caries and colon. Some important functional
oligosaccharides are described below.

Soybean oligosaccharide

Soybean oligosaccharide has once been regarded as an antinutritional factor and has been
reported to increase the incidence of diarrhea in rats. However, recent researches reveal that
soybean oligosaccharide has the potential as new functional ingredients in functional foods.
Soybean oligosaccharide has recently been found to be ―probiotic material‖ and have been
approved by the Food and Drug Administration as generally recognized as safe material in the
USA. Composition analysis indicates that soybean oligosaccharide was composed of
74 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

galactose (65.3%), mannose (15.6%), fructose (7.8%) and glucose (8.7%) [24]. Intake of 3~5
g soybean oligosaccharide per day is sufficient to enhance the proliferation of bifidobacteria.

Fructo-oligosaccharide

Fructo-oligosaccharides (FOS) are fructose oligosaccharides containing a single glucose


moiety. FOS are mainly composed of 1-kestose (GF2), nystose (GF3), and 1-β-fructofuranosyl
nystose (GF4), in which fructosyl units (F) are bound at the β(2 → 1) position of sucrose
molecule (GF). FOS occur naturally in various fruits and vegetables, such as bananas, onions,
chicory root, garlic, asparagus, barley, wheat, jícama, tomatoes, and leeks, but commercial
FOS are often prepared by transfructosylation from sucrose and chemical or enzymatic
hydrolysis of inulin.
FOS are about 0.4 and 0.6 times as sweet as sucrose and have been used in the
pharmaceutical and food industries as functional sweeteners. FOS with low polymeric grade
have better therapeutic properties than those with a high polymeric degree. They present
properties such as low caloric values, non-cariogenic properties, decreased levels of
phospholipids, triglycerides and cholesterol, help gut calcium and magnesium absorption, and
are used as prebiotics to stimulate the bifidobacteria growth in the human colon [25]. Figure
3-37 illustrates the structure of several FOS.

Xylo-oligosaccharide

Xylo-oligosaccharides (XOS) consist of 2~7 xylose residues connected through by the β


(1→4) glycosidic bond. XOS are about 40 times as sweet as sucrose. XOS have good thermal
stability and are not decomposed in acidic conditions (pH2.5~7) when heated. XOS can be
used in yogurt, lactobacillus drinks, carbonated drinks and other acidic beverages.
Commercial XOS consists mainly of xylose, xylobiose, and less amount of polymers with
DP greater than 3. Xylobiose is the main constituent of XOS and its content determines the
quality of XOS products. XOS can be prepared by the enzymatic (xylanase) hydrolysis of
xylan-rich materials, such as corncob, bagasse, cottonseed hull, and bran. Many fungi and
bacteria can produce xylanase, of which, the endoxylanase produced by Chaetomiu globosum
has been used in the industrial production of XOS.
XOS cannot be digested but can selectively activate bacterial reproduction within
intestines and hence are prebiological substances. It can obviously improve intestinal
microecological balance, proliferate bifid bacteria and gastric function.

Chito-oligosaccharides

Chito-oligosaccharides (COS) are oligomers of N-acetyl-D-glucosamine and D-


glucosamine connected through the β-1, 4 glycosidic bond. COS are water soluble in contrast
to chitosan and chitin. COS carry positive charge, which allows COS to bind to negatively
charged cell surface strongly. This property contributes to many physiological functions of
COS, such as antitumor, immunostimulatory, and anti-inflammatory. Besides, COS stimulate
the proliferation of Bifidobacteria bifidium and Lactobacillus sp. [26].
Carbohydrates 75

Figure 3-37. Structure of FOS.

Other oligosaccharides

Palatinose (6-O-α-D-glucopyranosyl-D-fructose), also referred as isomaltulose, is a


reducing disaccharide identified during the processing of sugar from sugar cane. It is
completely digested and provides the same caloric value as sucrose, but it is non-cariogenic
and digested much slower, leading not only to a low glycemic response but also to a
prolonged glucose supply, indicating its potential as a parenteral nutrient acceptable to both
diabetics and non-diabetics. Its ingestion selectively promotes the growth of beneficial
bifidobacteria amongst the human intestinal micro flora. It is more stable than sucrose, which
facilitates the maintenance of its sweetness and taste in fermented foods and beverages. It has
been suggested as a non-cariogenic alternative to sucrose, and as such is currently widely
used as a sugar substitute in foods. This disaccharide has a sweet taste and very similar
physical and sensory properties to sucrose [27].
Lactulose (4-O-β-D-galactopyranosyl-D-fructose) is a synthetic disaccharide and is also
termed isomerized lactose, because it can be prepared from the isomerization of lactose. It is a
Bifidus factor in nutrition and is a very important humanizing factor in infant formula and is
added to commercial infant formula products and various milk products. This sugar has
greater sweetness and solubility than lactose and if produced economically, it could widely be
used in baking and confectionery applications [28].
76 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Starch and Glycogen

Starch
Starch is the major constituent of many foods and is the most important carbohydrate
source for human nutrition. Corn, wheat, potato, sweet potato, and rice are the most important
materials for starch production. Table 3-8 lists the contents of starch in some crops.
Starch can be divided into two fractions: amylose and amylopectin. Natural starches are
the mixture of amylose (10~20%) and amylopectin (80~90%). The percentages of amylose
and amylopectin in starches derived from different sources are shown in Table 3-9.
Gelatinization does not occur easily in starches with high amylose contents and the
gelatinization temperature can reach up to 100 °C. However, gelatinized amylose is unstable
and is prone to aging in contrast to the high stability of gelatinized amylopectin.
Due to the unique physicochemical properties and nutritional functions, starches are of
incomparable importance to human beings. Starch and its derivatives have been widely used
as thickening agent, bonding agent and stabilizing agent and as the materials of pudding,
soup, sauce, vermicelli, infant foods, pie, and mayonnaise.

Chemistry structure

Amylose is a linear polymer of α-D-glucopyranosyl residues connected through the 1→4


glycosidic bond.
The number of repeated glucose subunits is usually in the range from 300 to 3000, but
can be up to many thousands. For example, the polymerization degree of wheat starch is in
the range 500~6000, while in potato it can rise up to 4500. Amylose also contain few α(1→6)
bonds, accounting for 0.3%~0.5% of total glycosidic bonds.
Based on X-ray diffraction diagrams, native starches can be divided into types A, B, and
C. An additional form, called the V-type, occurs in swollen granules. While types A and B are
real crystalline modifications, the C-type is a mixed form. The A-type is largely present in
cereal starches, and the B-type in potatoes, amylomaize, and in retrograded starches. The C-
type is not only observed in mixtures of corn and potato starches, but is also found in various
legume starches [29].

Table 3-8. Contents of starch in some materials (%)

Species Content Variety Content


Unpolished rice 73 Potato 16
Corn 70 Wheat 66
Barley 40 Sorghum 60
Kidney bean 49 Buckwheat noodles 72
Sweet potato (fresh) 19 Pea 58

Table 3-9. Percentages of amylose and amylopectin in some starches (%)

Starch Amylose Amylopectin Starch Amylose Amylopectin


High amylose corn starch 50~ 85 15~50 Potato starch 21 79
Corn starch 26 74 Cassava starch 17 83
Waxy corn starch 1 99 Wheat starch 28 72
Carbohydrates 77

Type B amylose shows the left-hand double helices structure, which are packed in a
parallel arrangement. Hydroxyl groups are located outside of the chains and the double helix
is stabilized by the hydrogen bridges between amylose molecules. The internal channel of the
helix is hydrophobic and hence can enclose only hydrophobic compounds, such as lipids. The
A-type is very similar to the B-type, except that the central channel is occupied by another
double helix, as shown in Figure 3-38.
Amylopectin is the highly branched polymer of glucose. The backbone chain is
composed of glucose units linked in a linear way with α(1→4) glycosidic bonds and
branching takes place through α(1→6) bonds. In amylopectin, α(1→6) bonds account for
4%~5% of total glycosidic bonds.

Figure 3-38. Unit cells and arrangement of double helices (cross section) in type-A amylose (left) and
type-B amylose (right) [29].

Due to the presence of multiple linear branches, amylopectin has multiple reducing ends
and are hydrolyzed by enzymes faster than amylose. The branches of amylopectin are
arranged in parallel or double helix, as shown in Figure 3-39.
Amylopectin contributes to the crystalline structure of starch granules. The molecular
weight of amylopectin ranges from 5×107~5×108. The ratio of amylopectin in starches
generally exceeds 75% and the value can reach up to 99% in some species, such as waxy corn
starch, as shown in Table 3-9. Starches derived from potato also contains a high phosphorus
content (0.06~0.1%). Hence, potato starches are slightly negatively charged and swell more
rapidly in warm water due to coulombic repulsion.

Gelatinization

Starch granule: Starches are present as granules in plant cells. Starch granules can be
round, oval, and polygonal and their sizes range from 0.001~0.15 mm, depending on the plant
78 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

species, of which, potato starch granules have the largest size and cereal starch granules have
the smallest size, as shown in Figure 3-40. Polarization microscopy observation and X-ray
diffraction find double refraction and X-ray diffraction in starch granules, indicating the
presence of crystalline structure alternate layers of crystal region and amorphous region
(Figure 3-41 I). About 70% of the mass of a starch granule is regarded as amorphous and
about 30% as crystalline.
The amorphous regions contain mainly amylose and the crystalline regions consist
primarily of amylopectin. Amylose can enclose fatty acids and hydrocarbons due to the
presence of the hydrophobic internal channel and the complexes are termed inclusion
complexes. Amylose occur as double helices as in starch granules. Amylose and amylopectin
are arranged radially in starch granules, as shown in Figure 3-41 II.

Figure 3-39. Structural models (I, II) for amylopectin with parallel double helices. III is an enlarged
segment of I or II.
Carbohydrates 79

Figure 3-40. Shapes of starch grains in electron microscope (×1200).A, Green bean starch (mean grain
size: 0.016nm); B, Potato starch (mean grain size: 0.049nm; C, Common corn starch (mean grain size:
0.013nm); D, Sweet potato starch (mean grain size: 0.017nm).

Figure 3-41. Sketch of crystallization section and amorphous section of starches (I), and radial shape of
amylose and amylopectin in starches (I, II).

Gelatinization: Due to the inter-molecular hydrogen bonding, starch is not soluble in cold
water in spite of the abundance of hydroxyl groups. When the suspension is heated, the
vibration of starch molecules increases and the hydrogen bonding between molecules is
disrupted. As a result, more hydroxyl groups are exposed and starch-water hydrogen bonding
is formed instead. With the diffusion of water into starch granules, many long chains are
separated and the confusion degree increases markedly. Meanwhile, the number and size of
80 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

crystal regions reduce significantly. When the suspension is further heated, the swelling
becomes reversible. In this case, random coils are observed in amylopectin due to hydration
and the ordered structure of starch is destroyed. This process is termed gelatinization and the
temperature at which irreversible changes occur is called the gelatinization temperature.
Starch gelatinization can be divided into three phases. In phase I, water diffuses into
granules and absorbs to the polar groups in amorphous regions below the gelatinization
temperature. The starch can restore to their original form if dehydrated. In phase II, when the
gelatinization temperature is reached, bulk water enters starch granules and the granules swell
significantly. Due to the increase of volume, the fraction between swollen starch increases
and the suspension becomes viscous, which can be followed by Brabender amylograph. In
this phase, water molecules enter the microcrystalline areas and the original arrangement is
disrupted. When the temperature further increases, the viscosity of the suspension increases
accordingly. In phase III, the swollen starch granules are disintegrated and the suspension
viscosity drops sharply, as shown in Figure 3-42. It could be seen that the shape of the curve
varies greatly for different starches.
The gelatinization of starch is affected by the following factors:
Water activity: Gelatinization occurs readily in medium with high water activity. The
presence of high concentration of sugars significantly suppresses gelatinization, because the
molecules can compete for water with starch.


6ISCOSITY"RABENDERUNITSX 








   
4IMEMIN

Figure 3-42. Gelatinization properties of various starches. Brabender viscoamylograph. 40 g starch/460


ml water, temperature programming: start at 50 °C, heated to 95 °C at a rate of 1.5°C/min. Held at 95°C
for 30 min – potato, - - - waxy corn, −−− normal corn, and ••• amylomaize starch.
Carbohydrates 81

Helix amylose

Fatty acid

Figure 3-43. Scheme of the lipid-starch inclusion complex.

Starch structure: Starches with higher ratio of amylose is less susceptible to


gelatinization and their gelatinization temperature might reach up to over 100 °C.
Salts: Salts in high concentrations suppress the gelatinization of starch, but those in low
concentrations have no the inhibitory effect, except potato starch, because salts affect the
charge carried on phosphorus groups.
Lipids: Lipids can be enclosed inside the helix of starch (Figure 3-43). The enclosed
lipids cannot be easily removed from the helix and can prevent the diffusion of water into
starch granules. Hence, any lipids that can complex with starch can prevent starch swelling
and gelatinization. The addition of C16~18 monoacylglycerol increases the gelatinization
temperature, reduces the gelation temperature, and weakens the gel strength.
pH: When the medium pH is lower than 4, starch is hydrolyzed to dextrins, which
reduces the suspension viscosity. Hence, crosslinked starch instead of natural starch should be
preferred as thickening agent in acidic foods. Starch gelatinization is not affected in the pH
range 4~7. Starch gelatinization occurs rapidly in medium with pH greater than 10, but such
high pH is not found in foods.
Amylase: In the initial phase of gelatinization, starch granules start to absorb water and
swell and amylase is not inactivated yet. Amylase can hydrolyze starch and accelerate
gelatinization. This is why new rice is gelatinized more easily than old rice, because the
former contains higher amylase activity.

Retrogradation

When a hot starch paste is cooled down, the suspension generally transforms to a
viscoelastic gel and starch becomes insoluble. The process of soluble starch becoming
insoluble is called retrogradation. The presence of conjunction zones in the gel indicates that
retrogradation is actually the recrystallization of starch molecules. The qualities of many
foods are deteriorated during retrogradation. For example, the staling of breads and the loss of
viscosity of soups and sauces are partially caused by starch retrogradation.
The retrogradation of starch is influenced by the following factors:
Amylose to amylopectin ratio: Amylose is more susceptible to retrogradation due to its
linear structure, of which, amylose with DP of 100~200 is the most resistant to retrogradation.
Starch concentration: Retrogradation occurs readily in high starch concentrations due to
frequent molecular collision, but water content of less than 10% hinders the process. The
highest retrogradation rate is generally found in the water content of 30~60%.
Presence of inorganic salts: Inorganic salts hinder the repositioning of starch molecules.
The hindering effects of some common ions are SCN-, PO43-, CO3-, I-, NO1-, Br-, CI-, Ba2+,
Ca2+, K+, and Na+ in the decreasing order.
82 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Medium pH: Starch undergoes retrogradation readily in the pH range 5~7 and the process
is inhibited in alkaline or acidic solutions due to charge repulsion.
Temperature: The optimum temperature for starch retrogradation is 2°C~4°C and
retrogradation does not occur in temperatures above 60°C or below -20°C.
Cooling speed: When starch paste is cooled slowly, starch molecules have sufficient time
to align and retrogradation occurs readily. If the paste is cooled rapidly, the water in the paste
crystallizes quickly, which prevents the approaching of starch molecules. Hence,
retrogradation occurs slowly in rapid cooling.
Presence of other ingredients: Lipids and emulsifying agents prevent retrogradation.
Compounds such as glyceryl monopalmitate (GMP), other monoglycerides and their
derivatives, and sodium stearoyl 2-lactylate (SSL) are often added into doughs of bread and
other baked goods to prevent starch retrogradation and increase shelf life.

Hydrolysis of starch

Starches can be randomly hydrolyzed by acids and enzymes. Under mild conditions,
starch is only partially hydrolyzed by acids. This process is called thinning and the products
are termed acid-modified or thin-boiling starch. Acid-modified starch has increased gel
transparency and strength and is less susceptible to retrogradation. Acid-modified starch has
wide applications and can be used as film forming agent and adhesive agent in products such
as pan-coated roasted nuts and candy. Besides, they are also used as encapsulating agents and
flavor carriers.
Enzymatic hydrolysis has been used in the production of commercially important syrups.
In the production of high-fructose corn syrup, corn is firstly hydrolyzed by α-amylase and
glucoamylase to obtain high purity D-glucose. D-glucose isomerase is then added and D-
glucose is converted to D-fructose. The final product is the mixture of 58% D-glucose and
42% D-fructose. High-fructose corn syrup is commonly used as sweetener in soft drinks.
The degree of hydrolysis of starch is measured by dextrose equivalency (DE), which is
defined as the percentage of reducing sugar in the syrup. DE is related to the degree of
polymerization and is calculated in the following equation:

(1)

Glucose polymers with DE less than 20 are defined as maltodextrins and those with DE
in the range 20~60 are termed corn syrup. Table 3-14 lists the functional properties of some
hydrolysis products of starch.

Modified starch

Physical, chemical and biochemical modifications have been implemented to enhance the
functional properties of starches and modified starches have gained wide applications in the
food, pharmaceuticals, and chemical industry. Important modified starches that are widely
used in the food industry are described below.
Low viscosity starch: Starch of this type is also termed acid-modified starch. When starch
is exposed to acid below the gelatinization temperature, hydrolysis occurs only in the
Carbohydrates 83

amorphous region and the crystal region nearly remains intact. The modified starch produced
under such conditions is not soluble in cold water but is readily soluble in boiling water.
Compared with native starch, low viscosity starch has decreased viscosity and gel strength of
hot paste and elevated gelatinization temperature. Low viscosity starch can be used as
thickening and film forming agents.
Pre-gelatinized starch: This product is obtained by heating starch suspension above the
gelatinization temperature and then dried with drum drying, spray drying or extrusion. Pre-
gelatinized starch can dissolve and form gel in cold water. Pre-gelatinized starch can be used
in elder and infant foods, surimi products, ham, sausage, and bakery foods. Besides, pre-
gelatinized starch has gained applications in cooking-free instant foods due to its solubility in
cold water.

Table 3-14. The functional properties of hydrolysis products from starch

Properties enhanced by greater Properties enhanced in products of less


hydrolysis A conversion B
Sweetness Viscosity production
Hygroscopicity and humectancy Body formation
Freezing point depression Foam stabilization
Flavor enhancement Sugar crystallization prevention
Fermentability Ice crystal growth prevention
Browning reaction
Note: A high DE syrups; B low DE syrups and maltodextrins.

Etherified starch: All the three free hydroxyl groups in each D-glucopyranose unit can be
etherified. Hydroxyethyl starches of low degree of substitution (DS) have reduced
gelatinization temperature, increased swelling rate, and lowered tendency of pastes and gels
to retrogradation. Hydroxyalkyl starches, such as hydroxypropyl starch, can be used in salad
dressings, pie fillings, and other foods as thickening agent.
Esterified Starch: Starch can be esterified by acidic orthophosphates, acid
pyrophosphates, and tripolyphosphates to produce various starch esters. Compared with
native starch, monostarch phosphates gelatinize in lower temperatures and those with DS
greater than 0.07 can swell in cold water and have increased paste viscosity and transparency,
and decreased retrogradation. These properties are quite similar to those of potato starch,
which has high phosphorus content.
Because monostarch phosphates have good freezing-thawing stability, they are preferred
in frozen foods, such as frozen broth and frozen cream pie, as thickening agent to unmodified
starch.
Starch can also be esterified by various organic acids, such as acetic acid, long-chain fatty
acids (C6~C26), succinic acid, adipic acid, and citric acid. These esters are superior in
thickening and paste transparency and stability to native starch and can be used in bakery
products, soup powder, sauce, pudding and frozen foods as thickening agent and stabilizer, as
well as in dehydrated fruit as protective agent and in flavor processing as encapsulating
material.
Crosslinked starch: Starch can react with sodium trimetaphosphate, phosphorus
oxychloride, epichlorohydrin, and acid anhydride to yield crosslinked derivates. Crosslinking
84 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

prevents starch granule swelling and increases the stability against heating. Crosslinking by
phosphorus increases the stability of swollen starch granules, but the paste of distarch
phosphates is opaque in contrast to that of monostarch phosphate.

Table 3-15. Comparisons of the properties of native and modified starches

Starch Amylose/Amylopectin Gelatinization Properties


ratio temperature range (°C)
Common 1:3 62~72 Poor freezing-thawing stability
starch
Waxy starch 0:1 63~70 Low tendency of paste to
retrogradation
High-amylose 3:2-4:1 66~92 Less birefraction of granule
starch than common starch granules
Acid- Variable 69~79 Reduced viscosity of hot paste
modified
starch
Hydroxyethyl Variable 58~68 (DS0.04) Increased paste transparency
starch and decreased retrogradation
Monostarch Variable 56~66 Reduced gelatinization
phosphate temperature and retrogradation.
Crosslinked Variable Higher than unmodified Reduced peak viscosity and
starch starch, depending on the increased paste stability
degree of crosslinking.
Acetylated Variable 55~65 Increased paste transparency
starch and stability

Starches with high degree of crosslinking are very stable against high temperature, low
pH and mechanical vibration and their gelatinization temperature is proportional to the degree
of crosslinking. Some starches with high degree of crosslinking do not swell even in boiling
water.
Crosslinked starches are mainly used in infant foods, salad dressing, fruit pie filling and
cream-style corn as thickeners and stabilizers. They also provide resistance to gelling and
retrogradation, show good freeze-thaw stability, and do not undergo syneresis or weep on
standing.
Oxidized starch: Starch is hydrolyzed and oxidized when its aqueous suspension is
incubated with sodium hypochlorite below the gelatinization temperature and the carboxyl
group occurs once every 25 to 50 glucose residues in the resulting oxidation products.
Oxidized starch has relative low viscosity even in high concentrations and is used as
thickening agent in salad dressing and mayonnaise. Compared with low viscosity starch,
oxidized starch is less susceptible to retrogradation or forming opaque gel.
Table 3-15 lists the nature of a variety of starch before and after modification.

Glycogen
Glycogen is also called animal starch and is the major storage carbohydrate in muscle and
liver tissue of animals. Because glycogen accounts for only 0.02%~0.1% of fresh animal
Carbohydrates 85

tissues, it is less important for the food industry than starch. Glycogen is much branched than
starch and is similar to amylopectin in structure.

Cellulose and Hemicellulose

Cellulose
Cellulose is the major component of plant cell walls and often associates with
hemicellulose, pectin, and lignin. The mode and degree of their association significantly
affect the texture of plant-derived foods. Cellulose cannot be digested by the enzymes in
human digestive tract and hence is a good source of dietary fiber.
Cellulose is a linear homopolysaccharde consisting of D-glucopyranose units that are
connected through β-D-1,4-glycodic bonds. Native cellulose contains both crystal and
amorphous zones, of which, amorphous zone is more susceptible to solvent and chemical
reagent action. This difference has been used in the preparation of microcrystalline cellulose,
in which the amorphous zone is hydrolyzed by acids and the acid-resistance crystal zone is
remained. The molecular weight of microcrystalline cellulose ranges from 30~50kDal and its
tradename is avieol. Microcrystalline cellulose is insoluble in water and is often added to low-
calorie foods as fillings and rheology control agents.
The degree of polymerization (DP) of cellulose varies with the plant origin and ranges
from 1000~14000. Due to the large molecular weight and the presence of crystal structures,
cellulose is insoluble in water and its swelling power or the ability of absorb water is poor or
negligible.

Carboxymethyl cellulose (CMC)

CMC is the most widely used derivative of cellulose. It is obtained by treating alkaline
cellulose with chloroacetic acid. The DS of commercially important CMC ranges from
0.3~0.9 and DP from 500 to 2000.
CMC has long and rigid chains and carries positive charge. CMC solution is viscous and
stable due to the electrostatic impulsion. However, these properties depend on the DS and DP
of the product. Low-DS (≤ 0.3) CMC is insoluble in water but soluble in alkaline solutions,
while high-DS (> 0.4) are water soluble. Besides, the solubility and viscosity are also affected
by medium pH.
CMC with DS of 0.7~1.0 is used to increase the viscosity of foods. Their aqueous
solutions exhibit the characteristics of non-Newtonian fluid and the viscosity decreases with
temperature elevation. CMC solution is soluble in pH range 5~10 and is most stable in pH
7~9. Monovalent cations form soluble complex with CMC, divalent cations reduces CMC
solubility, whereas trivalent cations can cause gelling or precipitation of CMC.
CMC can be used to improve the solubility of such food proteins as gelatin, casein, and
soybean protein by complexing with these molecules. CMC can maintain the stability of the
dispersion system of proteins even in their isoelectric points.
Due to the excellent rheologic properties, safety, and indigestibility, CMC is added to
jellies, paste fillings, spreadable process cheeses, salad dressings, and cake fillings as binding
and thickening agent. Meanwhile, because CMC has strong water binding capacity, it is
86 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

widely used in ice creams and other foods to prevent ice crystal formation. CMC also
increases the volume and elongate shelf life of cakes and other bakery foods, improves the
mouthfeel of sucrose and prevent CO2 escape from low-calorie carbonated beverage.

Methylcellulose (MC) and hydroxypropyl methylcellulose (HPMC)

MC is the etherified derivative of cellulose and is prepared by treating alkaline CMC with
chloroform. The DS of commercially important MC lies in the range 1.1~2.2.
MC is characterized by its gelling properties. When a MC solution is heated, the initial
viscosity drops with rising temperature and then increases sharply. This can be explained by
the breakdown of hydration layer around MC molecules upon heating, which increases the
hydrophobic interactions between MC molecules. Electrolytes such as NaCl and
nonelectrolytes such as sucrose and sorbierite reduce the gelling temperature of MC by
competing for water molecules. MC cannot be digested by human body and is a calorie-free
polysaccharide.
HPMC is prepared by incubating cellulose with methyl chloride and cyclopropane in
alkaline solutions. The DS of commercial HPMC ranges from 0.002 to 0.3.
The initial viscosity of HPMC solution decreases with temperature elevation and the
formation of gel is reversible at specific temperature, which is similar to that of MC. The
gelling temperature and gel strength are related to the type of substation groups, DS and
concentration of the soluble gel. The hydroxypropyl group stabilizes the hydration layer of
HPMC and thereby increases the gelling temperature. Changing the proportion of methyl to
hydroxypropyl substituents can vary the jelling temperature within a wide range.
MC and HPMC increase the water retention and absorption capabilities of foods and
prevent excessive oil absorption in fried foods. MC can be added to functional foods as
fillings and dehydration and shrinkage inhibitors. The two derivatives are added to salad
dressings as thickening and stabilizing agents and to various foods as edible coatings and fat
substitutes.

Hemicellulose
Hemicelluloses occur along with cellulose in the cell walls of plant cells and consist of
multiple monomers in contrast to cellulose, such as xylose, mannose, galactose, rhamnose,
and arabinose in addition to glucose. The compositions of hemicelluloses vary with the plant
origin and tissue. For example, wheat and rye contain mainly arabinoxylans, while β-glucans
predominate in barley and oats.
In the food industry, hemicellulose is mainly added to bakery foods to increase the water
binding capability of flour, improve the kneading quality of dough by reducing the energy
required by kneading, and increase bread volume. Breads containing hemicellulose have
delayed hardening time compared with those without hemicellulose. Hemicellulose is an
important source of dietary fiber.
Carbohydrates 87

Pectin

Pectins are polymers consisting of D-galactopyranosyluronic acids connected through the


α-1,4-glycodic linkage. In addition to galacturonic acid, pectins might also contain rhamnose,
galatose, arabinose, and other sugars. Pectins are present in the cells and intercellular layers
of plants. Pectins originated from various sources differ mainly in their contents of methoxyl
groups or degrees of esterification (DE). The DE of pectins is defined as the percentage of the
number of esterified D-galacturonic acid residues among all D-galacturonic acid residues.
Pectins with DE greater than 50% are termed high-methoxyl pectin (HM) and those with
DE less than 50% are called low-methoxyl pectin (LM). Protopectins are the highly methyl
esterified and insoluble form of pectins and are found in immature fruits and vegetables.
Pectins with low DS are called pectinic acids and can be converted from protopectins by
the hydrolysis of protopectinase and pectin methyl esterase. Pectinic acids are either colloidal
or soluble in water depending on the degree of polymerization and degree of methyl
esterification, of which, water soluble pectinic acids are also referred as low-methoxyl pectin.
Pectinic acids can be completely hydrolyzed by pectin methyl esterase to pectic acids.
Various pectinases participate in the post-harvesting ripening of plants. During this
process, protopectinase converts protopectins to colloidal or soluble pectinic acids.
Pectinesterase removes methoxyl groups from pectins to produce poly-D-galacturonic acid or
pectic acid, which is then hydrolyzed by polygalacturonase to yield D-galacturonic acid units.
These enzymes work together in the ripening of fruits and are important for the texture
formation of fruits and vegetables.
Pectin is an important polysaccharide with applications in foods, pharmaceuticals, and a
number of other industries. Its importance in the food sector lies in its ability to form gel in
the presence of divalent ions such as Ca2+ or a solute at low pH. In the case of HM, gels are
formed only in low pH and high sugar concentrations. Generally, HM concentration 1%, pH
2.8~3.3 and sucrose concentration 58%~75% facilitate the gelation of HM. In HM, the cross-
linking of pectin molecules involves a combination of hydrogen bonds and hydrophobic
interactions between the molecules. Low pH suppresses the dissociation of carboxylic groups
and the loss of charges minimizes the electrostatic impulsion between HM molecules.
Meanwhile, sugars compete for water molecules and reduce the solvation of HM chains,
facilitating the hydrogen bonding between HM molecules and consequently their gelation.
HM gels can maintain their original properties even when heated at 100 °C.
The strength of pectin gels is positively proportional to the molecule weights and inter-
molecular association of pectins. Generally, the gelation time increases as the degree of
methyl esterification increases from 30%~50%, because the raise of carbomethoxy groups
increases the steric hindrance for hydrogen bonding between pectin molecules. Pectins with
degree of methyl esterification in the range 50%~70% have enhanced hydrophobic
interactions and therefore gel in shorter time. The gelling characteristics of pectins are the
function of degree of methyl esterification, as shown in Table 3-16.
In the case of LM, the presence of divalent ions, such as Ca2+, is a prerequisite for their
gelation and the ions function as the bridge between LM molecules. Increasing the
concentration of Ca2+, which is the only divalent cation allowed in the food industry,
increases the gelling temperature and gel strength of LM, which is similar to the role of Ca2+
in the formation of the egg-box structure in alginate gels. LM is not as sensitive to pH as HM
and can gel in a wider pH range 2.5~6.5. Though sucrose is not a prerequisite for LM
88 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

gelation, the addition of 10%~20% sucrose markedly improves the texture of gels. LM gels
are less rigid or elastic than common pectin gels, but sugars or plasticizing agents improve the
properties. LM pectin, since it does not require sugar for gelation, is used to make dietetic
jams, jellies, and marmalades.

Agar

Agar is a gelatinous polysaccharide extracted from some species of Rhodophyceae by a


hot water extraction process.

Structure and Properties


Agar is a linear heteropolysaccharide consisting of alternate β-D-galactopyranose and
3,6-anhydro-α-L-galactopyranose that are connected through 1→4 and 1→3 linkages
respectively.

Table 3-16. Effects of degree of methyl esterification on the gelling properties of pectin

Degree of methyl Gelling conditions Rate of


esterification pH Sugar concentration Presence of gelation
(%) divalent ions
>70 2.8~3.4 65% No High
50~70 2.8~3.4 65% No Low
<50 2.5~2.6 None Yes High

The hydroxyl groups on the chains are more or less esterified by methyl, sulfuric acid,
and pyruvic acid. Commercial agar is colorless or light yellow strips or powder. Agar is
insoluble in cold water, but dissolves slowly in hot water. The β-1,4-glycodic bonds can be
specifically hydrolyzed by the agarlytic enzymes derived from soil and marine organisms to
yield agar oligosaccharide. Figure 3-44 illustrates the structure of agarose.

Figure 3-44. Structure of agarose.


Carbohydrates 89

Figure 3-45. Molecular structures of the seven carrageenan classes.


90 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Application
Agar is a most potent gelling agent in the food industry and is widely used in candies,
puddings, ice cream, gelly, and many other foods as gelling agent. Besides, agar is
indigestible and is a good source of dietary fiber. In the fermentation industry, agar has been
widely used to encapsulate enzymes and microbes.

Carrageenan

Carrageenans are mixtures of several related galactans having sulfate half-ester groups
attached to the sugar units and are extracted from the red seaweeds (Rhodophyceae).
Carrageenans are important commercial hydrophilic gels.
Carrageenans are linear chains of D-galactopyranosyl units joined with alternating (1,3)-
D- and (1,4)-D-glycosidic linkages, with most sugar units having one or two sulfate groups
esterified to a hydroxyl group at carbon atoms C-2 or C-6. The molecular weights (MWs) of
carrageenan range from (1~5)×105 and the average MW of food-grade carrageenans is 2×105.
According to the positions of sulfate half-ester groups, carrageenans are divided into κ-, ι-, λ-,
μ-, υ-, θ-, and δ- classes. Figure 3-45 presents the structures of the 7 carrageenan classes.

Table 3-17. Properties of κ-, ι-, and λ- carrageenans

Item Condition κ-carrageenan ι-carrageenan λ-carrageenan


Solubility Hot water Soluble above 70°C Soluble above Soluble
70°C
Cold water Sodium salt soluble Na+ Ca2+ salt gives All salts soluble
salt soluble, From limited thixotropic
to high swelling of K+, dispersions
Ca2+ and NH4 salt
Hot milk Soluble Soluble soluble
Cold milk Insoluble Insoluble Disperses with
thickening
Cold milk Thickens or gels Thickens or gels Increased
(Tetrasodium thickening or
pyrophosphate) gelling
Concentrated sugar Soluble hot Difficulty soluble Soluble hot
solutions
Concentrated salt Insoluble cold and hot Soluble hot Soluble hot
solutions
Organic solvents Insoluble Insoluble Insoluble
Gelling Cations Hard gel with K+ Strong gel with Non-gelatinous
properties Ca2+
Gel type Rigid with syneresis Elastic without Non-gelatinous
syneresis
Effect of locust bean Synergetic Non-synergetic Non-synergetic
gum
Stability against Stable Stable Stable
neutral and alkaline
conditions
Stability in pH3.5 Hydrolysis, accelerated Stable hydrolysis
solutions upon heating; gel state
Miscibility Miscible with nonionic and anion surfactants, immiscible with cation surfactants
Carbohydrates 91

Structure and Properties


Commercial carrageenans are odorless, colorless or light yellow powder. Carrageenans
have strong gel forming ability and the gel is heat reversible, that is, formed gels can be
transformed to solutions by heating or vice versa upon cooling. All the carrageenans are
soluble in hot water or hot milk, but their potassium and calcium salts only absorb water in
cold water without dissolution. Carrageenans are not soluble in methanol, ethanol, propanol,
isopropanol, or acetone. Of the seven classes, κ-, ι-, and λ- carrageenans are commercially
important for the food industry and their properties are summarized in Table 3-17.

Applications in the Food Industry


Carrageenans have been used as nature food additives for hundreds of years. Carrageenan
utilization in food processing is based on the ability of the polymer to gel, to increase solution
viscosity and to stabilize emulsions and various dispersions. A level as low as 0.03% in
chocolate milk prevents fat droplet separation and stabilizes the suspension of cocoa particles.
Carrageenans prevent syneresis in fresh cheese and improve dough properties and enable
a higher amount of milk powder incorporation in baking. The gelling property in the presence
of K+ salt is utilized in desserts and canned meat. Protein fiber texture is also improved.
Protein sedimentation in condensed milk is prevented by carrageenans which, like κ-casein,
prevent milk protein coagulation by calcium ions. Carrageenans are also used to stabilize ice
cream and clarify beverages. Besides, because carrageenans are cheaper than agar, they have
replaced agar in some foods. The applications are summarized in Table 3-18.

Table 3-18. Applications of carrageenans in the food industry

Food Function Food Function


Ice cream Ice cream formation and Dessert gels Gelling
syneresis inhibition
Chocolate milk Coca suspension Fruit drinks Fruit particles suspension and gelling
Milk Smoothing and texture Bread Water retention increasing, staling
modifier delaying
Processed Syneresis inhibition, Pie Texture modification
cheese shape retention, and curd
formation reduction
Infant Fat and protein Seasonings Ingredient suspension and shape
formulations stabilization retention
Milk pudding Gelling agent and texture Canned foods Gelling and fat stabilization
modification
Milk shake Ingredients suspending Meats Syneresis prevention and bonding
and texture modification
Acidic dietary Texture modifier and Beer Clarification and stabilization
products

Alginate

Alginate, also termed algin or alginic acid, is an anionic polysaccharide obtained from
brown seaweeds. Commercial alginate is often available as sodium salt.
92 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Structure and Properties


Alginates are linear copolymers composed of -D-mannopyranosyluronic acid and -L-
gulopyranosyluronic acid units, which are joined by 1→4 linkages.
The ratio of the two sugars (mannuronic/guluronic acids, M/G) depends on the source
and is generally 1.5.
Pure alginates are colorless and amorphous and are insoluble in pure water, ethanol,
carbon tetrachloride or other organic solvents. Alginates can absorb much water and swell in
pH 5.8~7.5 to produce transparent and viscous solution. Dissolved alginates can be
precipitated by acid or calcium salts, which are the methods for the industrial production of
alginates. The presence of proteins, sugars, salts, glycerol, and trace amount of starch and
phosphates has no effect on the solubility of sodium alginate.
Alginate solutions are very viscous and the specific viscosity depends on alginate
molecular weight. Generally, the higher the molecular weight, the higher the viscosity of
alginate solutions. The viscosity of alginate solutions increases markedly with the increase of
alginate concentration. When alginate concentration exceeds 3%, solutions lose the fluidity.
High temperature reduces alginate viscosity. When the temperature increases by 1°C, the
viscosity decreases by about 3%. When the temperature exceeds 80°C, decarboxylation
reaction occurs and the viscosity drops dramatically. Alginates are liable to decreases of
degree of polymerization and viscosity due to the effects of temperature, light, metal ions, and
microbes during storage. Alginates are soluble in solutions with pH greater than 5.8. When
the pH decreases to below this value, the solubility decreases and gel is formed.
When a calcium salt is added to alginate sodium solution, the calcium ion replaces the
sodium ion and gelling occurs. Hence, alginate gels can be formed by adding either Ca2+ or
acids.

Applications in the Food Industry


Alginate is a powerful thickening, stabilizing and gel-forming agent. It has gained wide
applications in the food industry, as listed in Table 3-19.

Table 3-19. Properties of alginate and its major application in foods

Application Property utilized Alginate type


Cold drinks and snacks (ice cream) Thickening, hydratability, Sodium alginate, calcium
reaction with Ca2+ alginate, PGA1
Jams, mayonnaise, tomato paste, Gelling, thickening, and Sodium alginate, calcium
seasoning dressing emulsifying alginate, PGA
Noodles, instant noodle, macaroni, Gelling, thickening, acid PGA, sodium alginate
bread resistance
Jelly foods (meat jell, fish jell) Gelling PGA, sodium alginate
Wines (beer, white wine, fruit Foam stabilizing, PGA, sodium alginate
wine) coagulating, clarifying
Candies (cerealose, chocolate) Thickening, bonding Sodium alginate, calcium
alginate, PGA
Meat emulsion, fish emulsion Stabilizing, bonding Sodium alginate, PGA
1
Note: PGA, propyleneglycol alginate.
Carbohydrates 93

Chitin and Chitosan

Chitin is the second most abundant natural polysaccharide in the world and its annual
product is 1×1011 tones, which is only behind that of cellulose. It is also the most abundant
naturally occurring polysaccharide that contains amino sugars. Chitosan is the deacetylated
products of chitin and has improved solubility and functional properties. Their abundance,
combined with their specific properties and functions, impart them with wide applications in
the food industry.

Structure and Properties


Chitin is a linear polymer consisting of N-acetylglucosamine connected through β-1,4
linkage. The general formula of chitin is (C8H13NO5)n and the molecular weight is of the
magnitude 106. Chitin is white or gray amorphous powder and decomposes at 270°C. Chitin
is insoluble in water, ethanol, diluted acid or diluted alkaline solutions and is soluble in only
several solvents, including hexafluoroacetone, trichloroacetic acid-dichloropropane,
ketopyrrolidine-lithium chloride, and some chlorohydrins. Chitin can be hydrolyzed by
concentrated inorganic acids.
Chitosan is a linear polysaccharide composed of randomly distributed β-(1-4)-linked D-
glucosamine and N-acetylglucosamine (acetylated unit). It is obtained by the deacetylation of
chitin. Commercial chitosan is white or gray amphorous powder and decomposes at 185°C.
Chitosan is insoluble in water, diluted alkaline solutions or some diluted acid solutions, for
example, sulfuric acid, nitric acid, and phosphorous acid, but soluble in diluted organic acids
and some inorganic acids such as hydrochloric acid. Chitosan carries positive charges and is
hence a polar polymer.

Applications of Chitosan in the Food Industry

Antibacterial agent

Chitosan and its derivates have strong antibacterial activities. Chitosan carries positive
charges and can adhere to negatively charged membranes of bacteria, leading to the leakage
of intracellular proteins and other components.

Fruit preservative

Chitosan can prevent the spoilage and decay of peach, kiwi fruit, cucumber, pepper, and
strawberry and markedly increase their shelf lives. This is because chitosan has strong film
forming capability. The film formed hinders the diffusion of oxygen to fruits and the escape
of CO2 generated by respiration, but allows the release of ethylene.

Antioxidants

It has been reported that the addition of chitosan at 1% resulted in a 70% decrease in the
2-thiobarbituric acid (TBA) values of meat after 3 days storage at 4 °C. The mechanism by
which this inhibition takes place is believed to be related to the chelation of free iron released
94 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

from the hemoproteins of meat during heat processing or storage. This would in turn inhibit
the catalytic activity of iron ions [30].

Functional ingredients

After chitosan is absorbed, it can form indigestible complexes with triglycerides, fatty
acids, bile acid, and cholesterol. To complement for the bile acid that is associated with
chitosan, the liver increases the secretion of bile acid. Since bile acid is converted from
cholesterol in the liver, the content of cholesterol in liver and blood is deceased. Hence,
chitosan is an effective ingredient of functional foods.

Juice clarificant

Fruit juices become turbid after long-time storage due to the presence of negatively
charged pectin, cellulose, tannin, and pentosan. The positively charged chitosan can coagulate
these components by electrostatic attraction and hence is widely used in fruit juice
clarification. Chitosan has high affinity to polyphenolic compounds, such as catechin and
cinnamic acid. Chitosan has been added to grape wines to remove these components.
Chitosan-treated grape wines are dark golden yellow compared to the light yellow of
untreated products and hence have improved quality.

Water clarificant
Chitosan is more effective in removing polychlorobiphenyl (PCB) than active carbon.
Chitosan in combination with bentonite can effectively remove particles, color, and odor of
drinking water and that in combination with polysilicate, polyaluminosilicate, and ferric
chloride markedly reduces the COD and turbidity of water.

Applications of Chitooligosaccharides in Food Industry


Chitooligosaccharide (COS) is the partial hydrolysis products of chitin and chitosan and
its molecular weight is generally less than 104. COS has much better solubility than chitin and
chitosan and has been found to posses various physiological functions. COS has great
potential applications in the food industry.

Micro-ecological regulator

COS is a Bifidobacterium proliferative factor and can selectively enhance the growth of
intestine probiotics and inhibit the proliferation of harmful bacteria and formation of spoilage
substances.

Functional sweetener

Chitobiose and chitotriose have pleasant sweetness and are superior to sucrose in
thermostability. The two compounds cannot be digested by human body and are ideal
functional sweeteners for the diets of diabetic and fat patients.
Carbohydrates 95

Antibacterial agent

The antibacterial activity of COS is related to its molecular weight and medium pH. It is
generally recognized that COS of molecular weight 1500 has the highest antibacterial
activity. Medium pH affects the dissociation of amino groups and therefore influences the
antibacterial activity. COS is more effective in inhibiting microbial growth in acidic medium
than in neutral or alkaline medium. Short-chain COS, such as those with seven or more
monomers, can enter inside microbial cells and inhibit the biosynthesis of mRNA and
proteins by binding to DNA, thus killing the microbes.
COS is more effective in inhibiting the growth of molds in soy sauce than benzoic acid
and sodium benzoate without affecting the taste and color of soy sauce.

Preservative

COS has abundant free amino and hydroxyl groups and therefore has strong water
retention capability. The mixture of COS and high-molecular weight chitosan with strong
film forming ability has been used as fruit and vegetable coatings to prevent water
evaporation and Vc loss and elongate the shelf life. COS also significantly reduces the
spoilage rate of fruits and vegetables as chitosan.

Calcium absorption enhancer

COS of DP 3~7 reduces the excretion of fecal calcium and increases the fracture force of
rat thighbone in contrast to chitosan, which hinders the absorption of calcium.

Guar Gum and Locust Bean Gum

Guar gum and locust bean gum are important thickening polysaccharides and are widely
used in the food and other industries. Guar gum produces the highest viscosity of all natural
and commercial gums.
Guar gum is also known as guaran and occurs in the endosperm of the seeds of guar bean
(Cyamopsis tetragonolobus). It is composed of galactose and mannose and the backbone is a
linear chain of β-1,4-linked mannose residues to which galactose residues are 1,6-linked at
every second mannose, forming short side-branches. The structure of guar gum is shown in
Figure 3-46.
Guar gum readily absorbs water and is rapidly hydrated in cold water to produce highly
viscous and thixotropic fluids. The viscosity of guar gum solutions is affected by temperature
and the presence of other components.
Heating accelerates the dissolution of guar gum, but it can be decomposed in high
temperatures. Because guar gum yields very high viscosity even in low concentration, the
addition of guar gum in foods is generally less than 1%.
The aqueous solution of guar gum is neutral and its viscosity is not affected by pH.
Hence, guar gum is compatible with most food components. Ionic strength has little effects on
the viscosity of guar gum solutions, but the presence of high concentrations of sucrose
96 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

reduces the viscosity and delays the arrival of the peak viscosity. Wheat starch synergistically
improves the viscosity of guar gum solutions.
Guar gum prevents ice crystal growth and produces desired consistency, chewiness, and
heat stimulation resistance in ice cream. In cheese, guar gum is added to avoid syneresis. In
bakery foods, guar gum elongates the shelf life and reduces the water absorption of sucrose in
the glace of pastries.
Locust bean gum, also called carob gum, occurs in the seeds of carob tree (Ceratonia
siliqua). It structure is similar to that of guar gum, except that the galactose residues are
distributed randomly along the β-1,4-linked mannose backbone.
The ratio of mannose/galactose residues of locust bean gum ranges from 3:1 to 6:1 and its
molecular weight is about 31000. Due to the irregular distribution of galactose residues, some
mannose residues are exposed or naked without attached galactose residues. Due to this
difference, locust bean gum has different properties compared to guar gum. For example, the
viscosity of locust bean gum solutions is lower than that of guar gum solutions.
Locust bean gum is used as a thickener, binder and stabilizer in canned meats, salad
dressings, sausages, soft cheeses, and ice creams. It also improves the water binding capacity
of dough especially in the case of low-gluten flour. Locust bean gum can improve the
viscosity of other gelatinous polysaccharides. For example, the viscosity of the solution
containing 0.5% agar and 0.1% locust bean gum is 5 times higher than that of agar alone.

Gum Tragacanth

Gum tragacanth is a plant exudate collected from the plants of the Astragalus genus. The
gum has been used for over 2000 years as Arabic gum and is mainly produced in Iran, Syria,
and Turkey.
Gum tragacanth has a very complex structure and composition. When gum tragacanth is
suspended in water, one fraction is dissolved and this part is called tragacanthic acid.
Tragacanthic acid accounts for 60%~70% of gum tragacanth and consists of 43% D-
galacturonic acid, 10% fucose, 4% D-galactose, and 40% D-xylose and L-arabinose, of
which, the galacturonic acid residues form the backbone and the other residues are attached as
side chains. The molecular weight of tragacanthic acid is about 800000. The insoluble
fraction is termed bassorin, which contains 75% L-arabinose, 12% D-galactose, and 3% D-
galacturonase and L-rhamnose with molecular weight of 84000.

Figure 3-46. Structure of guar gum.


Carbohydrates 97

Gum tragacanth produces high viscosity and hence is used as thickening agents. Gum
tragacanth is used as a thickening agent and a stabilizer in salad dressings (0.4–1.2%) and in
fillings and icings in bakery goods. As an additive in ice creams (0.5%), it provides a soft
texture.

Microbial Polysaccharides

Microorganisms can produce multiple polysaccharides, of which, dextran and xanthan are
the most important for the food industry.
Dextran is an extracelluar polysaccharide produced by Leuconostoc mesenteroides,
Streptobacterium dextranicum, Streptococcus mutans and other species. Dextran is composed
of glucose units, of which, the monomers in the straight chain are connected through α-1,6
linkages, while branches begin mainly from α-1,3 linkages (Figure 3-47), with part 1,4- and
1,2- linkages. The type and number of the glycosidic linkages vary with the source. The
dextran produced by Leuconostoc mesenteroides NRRL B512 contains 95% 1,6- linkages and
the remaining 5% are 1,3- and 1,4- linkages.
Dextran is used in confections to improve viscosity and water retention and inhibit sugar
crystallization. In chewing gum and soft sweets, dextran is added as gelling agent. Dextran
inhibits ice crystal formation in ice creams and provides desired consistency and mouthfeel
for puddings.

Figure 3-47. Structure of dextran.

Figure 3-48. Structure of xanthan.


98 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Xanthan is secreted by multiple species of the Flavobacterium genus and the commercial
xanthan is produced by Xanthomonas campestris. Xanthan can be considered as derivative of
cellulose.
On an average, every second glucose residue bears in the 3-position a trisaccharide of the
structure β-D-Manp- (1→4)-β-D-GlcpA(1→2)-α-D-Manp as the side chain. The mannose
bound to the main chain is acetylated in position 6 and ca. 50% of the terminal mannose
residues occur ketalized with pyruvate as 4,6-O-(1-carboxyethylidene)-D-mannopyranose
(Figure 3-48). The molecular weight of xanthan is greater than 2×106 Dal.
Xanthan is readily soluble in both cold and hot water and provides high viscosity even in
low concentrations, but the jellification is weakened in high xanthan concentrations. Xanthan
solutions are pseudoplasitc fluids and exhibit obvious shear thinning. Xanthan is compatible
with most edible salts and edible acids and can coexist with them in foods. Xanthan interacts
with guar gum to give rise to a synergistic increase in solution viscosity and with locust bean
gum to produce a heatreversible gel.
Xanthan is widely used to stabilize aqueous dispersions, suspensions, and emulsions.
Xanthan is added to juices and canned foods as suspending and stabilizing agents. In starch
gels, the addition of xanthan gum substantially improves their freeze-thaw stability and
reduces syneresis. Due to the high stability of xanthan gels, xanthan can be used in seasonings
with high salt contents or acidity.

Glucomannan

Glucomannan is slightly branched polymer consisting of β-(1→4)-linked D-mannose and


D-glucose in a ratio of 1.6:1. Short side chains of 11-16 monosaccharides occur at intervals of
50-60 units of the main chain attached by β1→3 linkages. Some native glucomannan are
partially acetylated at C-2 and C-3 of certain of mannose residues [31]. The weight-average
molecular weights range from 105~106 and depend on source.
Glucomannan is soluble in water to produce highly viscous pseudoplastic fluids. In
alkaline conditions, glucomannan is deaceylated and aggregates to form the three-
dimensional network and strong thermo-irreversible gel [32]. However, glucomannan can
interact with xanthan to yield thermo-reversible gel.
Glucomannan has strong hydrophilicity, gelation capacity, and film-forming ability.
Glucomannan is added to jellies, jams, confections, dietary products, ice creams, meat
products, and breads as thickening and stabilizing agents.

Gum Arabic

Gum Arabic is extracted from the exudate of various Acacia species, primarily Acacia
senegal. Gum Arabic is a proteoglycan and its molecular weight lies in the range
260000~1160000. The glycan fraction accounts for 70% of the total gum and consists of L-
arabinose, D-galactose and D-glucuronic acid in the molar ratio of 3.5:1.1:2.9:1.6. The
backbone of gum Arabic consists of β-D-galactopyranosyl residues linked by 1→3 bonds and
side chains are attached at position 6 (Figure 3-49). Gum Arabic is present as acidic or neutral
salts and the counter ions are Ca2+, Mg2+, and K+. The protein fraction accounts for 2% of the
Carbohydrates 99

total gum, but the value can reach up to 25% in some species. The glycan fraction is
covalently connected to the hydroxyproline and serine residues of the protein fraction.
Gum Arabic is readily soluble in water and provides low viscosity, but the viscosity
increases dramatically in high gum Arabic concentrations. This property is different from
other polysaccharides, which can provide high viscosity even in low concentrations. The
solubility of gum Arabic reaches up to 50% by weight. Arabic gum solutions with
concentration less than 40% are Newtonian fluids, but those with higher concentrations
exhibits the pseudoplastic behavior.
In the presence of ions, the viscosity of gum Arabic solutions varies with pH and the
highest viscosity is obtained at pH 6~8. The viscosity reduces proportionally when the
valence and concentration of ions increases. Gum Arabic is compatible with most gums,
except gelatin and sodium alginate.

Figure 3-49. Structure of the glycan fraction in gum Arabic.


100 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Gum Arabic is used as emulsifier and stabilizer. It retards sugar crystallization and fat
separation in confectioneries and large ice crystal formation in ice creams, and can be used as
a foam stabilizer in beverages. Gum Arabic is also applied as a flavor fixative in the
production of encapsulated, powdered aroma concentrates. For example, essential oils are
emulsified with gum Arabic solution and then spray-dried. In this process, the polysaccharide
forms a film surrounding the oil droplet, which protects the oil against oxidation and other
changes.

DIETARY FIBER
Dietary fibers (DF) were once considered as nonnutritive crude fibers and unimportant
for human diets. However, with the rapid improvement of social economy and people's living
standards, people‘ dietary structure changes greatly. The proportion of plant-derived foods in
diets decreases obviously and that of animal-derived foods containing high calorie, high
protein content and high fat contents increases significantly instead, which impairs the
nutrition balance of the diets. Due to the imbalance, the incidences of adiposity, hypertension,
diabetes, cancer, and cardiovascular diseases keep rising. The emergency of these diseases
has been contributed to the decreased intake of DF and DF has been widely accepted as the
seventh essential nutrient.

Structure and Properties

Definition
Various definitions of dietary fibers have been proposed. In 1987, FDA and WHO
defines dietary fiber as ―the material isolated by AOAC method 985.29‖. In 1998, dietary
fiber was defined by a committee of dietary fiber experts of the American Association of
Cereal Chemists (AACC) as ―the edible parts of plants or analogous carbohydrates that are
resistant to digestion and absorption in the human small intestine with complete or partial
fermentation in the large intestine. Dietary fiber includes polysaccharides, oligosaccharides,
lignin, and associated plants substances. Dietary fibers promote beneficial physiological
effects including laxation, and/or blood cholesterol attenuation, and/or blood glucose
attenuation‖. The definition proposed by AACC has been widely accepted in the world.

Classification

By Solubility
Based o solubility, DF is subdivided into soluble DF (SDF) and insoluble DF (IDF).
SDF is soluble in warm and hot water and absorb water to gel. Dissolved SDF can be
precipitated by 4 volumes of ethanol. SDF mainly includes pectins, gums (gum Arabic, guar
gum, locust bean gum, and agar), mucilages, and some hemicelluloses. IDF is insoluble in hot
water and is the important component of plant cell wall, including cellulose, hemicellulose,
ligin, protopectin, vegetable wax and some animal-derived compounds, such as chitin and
chitosan.
Carbohydrates 101

SDF and IDF have different physiological and health functions. IDF cannot be
metabolized by human body, but can absorb water as it moves through the digestive system,
thus easing defecation. SDF can be partially fermented by the microorganisms inhabited in
the colon and produce active byproducts. SDF protects against cholesterol gallstone [33],
increase heavy metal evacuation, reduce cholesterol levels in serum and liver, inhibit
postprandial blood glucose increase, and prevent hypertension and heart disease. IDF
increases defecation and prevents obesity, laxation, and intestinal cancers. The ratio of IDF to
SDF markedly affects the physiological functions of DF.

By Source
DF can be obtained by extraction from plants (including algae) and animals or by
synthesis, of which, plants are the major source at present. Chitin and chitosan are typical
representatives of animal-derived DF. Dextran is a water soluble DF. In addition to
microorganism, it is also synthesized from dextrin by transglucosylation of dextrin dextranase
(EC 2.4.1.2) [34].

Physichemical Properties

Solubility and viscosity

DF molecules with ordered and less branched structures have strong inter-molecular
bonding capacities and thereby poor solubility. For example, the linear cellulose is insoluble,
but pectins with irregular structure are readily water soluble. Insoluble DF can be converted
to soluble DF by high temperature, high pressure, or shearing force. Conversion of IDF to
SDF is an important measure to improve DF quality. SDF are viscous upon ingestion and can
complex with heavy metals, fats, and cholesterol.

Water retention

Most DFs contain abundant hydrophilic groups and can absorb much water. The water
retention capacity of DFs varies with the source, composition, structure, preparation method,
and particle size. Generally, cellulose-based DFs have lower water retention capacity
compared with other DFs. Too low particle sizes reduce the index. Besides, high temperature
treatment, cooking, and enzymatic hydrolysis could change the physical properties of DFs
and thereby influence the water retention capacity.

Organic compounds absorption

The abundant active groups on the surface of DF can absorb bile acid, cholesterol, and
mutagens in the intestines of human body and therefore affects their metabolism and
adsorption.

Cation binding and ion exchange


102 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

Some DFs contains carboxyl groups, hydroxyl groups, amino acids, and some other
active groups. Various cations, such as Ca2+, Fe2+, Zn2+, Cu2+, and Pb2+, can bind to these
groups or exchange with the ions attached to these groups. The binding or exchange is
reversible and changes the pH, osmotic pressure, and redox potential of the intestine and
provides a buffering environment that is beneficial for digestion and absorption. Cations with
high polarizability, such as Pb2+, are preferred in absorption by DF. Hence, DF has the
function of removing toxic metals. The ions bound to DF can exchange with Na+ and K+. The
exchange reduces the Na+/K+ ratio in the blood and consequently lowers the blood pressure. It
should be noted that DF also reduces the adsorption of some beneficial elements. Hence, it is
necessary to supplement some minerals in DF-containing foods to avoid mineral imbalance.

Prebiotics

DFs are only partially fermented in the colon. DFs are the nutrition source of the
microbes in the colon and support the growth of multiple bacteria, of which, most are
beneficial for human body. Besides, DFs can induce the substantial propagation of aerobes
and thereby reduces the incidence of some cancers.

Bulking effect

DFs have markedly increased volume after swelling and can cause satiety. Besides, the
presence of DF affects the digestion and absorption of other food components and delays the
feeling of hunger. Hence, DFs are effective in preventing adiposity.

Metabolism

As mentioned in the definition, DFs cannot be digested in the mouth, stomach, or small
intestine and only partially fermented by the microbes in the large intestine. The degree and
rate of decomposition are associated with the solubility, chemical structure, particle size, and
ingestion mode of DF. Soluble DFs, such as pectin and seaweed gums, are readily digested in
the large intestine, but insoluble cellulose cannot be easily utilized by intestinal
microorganisms. Some of the metabolites, such as fatty acids, can be absorbed by human
body as energy source and some metabolites have important physiological functions.

Safety Aspects

Although DFs have multiple beneficial effects on human body, excessive intake of DFs
can cause discomfort and affect the absorption of fats, proteins, and essential elements. It
should be mentioned that these effects vary with DF composition and individuals.
Some IDFs, such as guar gum, can be fermented by intestinal microorganisms to produce
volatile fatty acids, carbon dioxide, and methane, which can cause abdominal distension. The
filling effect of DFs reduces the adsorption of proteins, fatty acids, and carbohydrates and the
increased viscosity of gastrointestinal fluids prevents the contact between these substrates
Carbohydrates 103

with digestive enzymes. Hence, DFs reduce their bioavailability. DFs can bind metal ions and
can reduce their absorption.
Some DFs interfere with the absorption of vitamins. It is reported that konjac mannan
reduces fat-soluble vitamin absorption by removing bile acids, but does not reduce fat-
insoluble vitamin absorption in the intestine [35].
No uniform standards have been formulated for the daily intake of DFs due to the
complexity of DFs and individual differences. The daily intake recommended by FDA is
20~35g for adults and that in Japanese is 20 g or more. The Chinese Nutrition Association
enacted the Chinese dietary intake references in 2000 and suggests that the daily dietary fiber
intake of adults is 30.2 g.

REFERENCES
[1] Terta, MG; Blekas, G; Paraskevopoulou, A. Retention of selected aroma compounds by
polysaccharide solutions: A thermodynamic and kinetic approach. Food Hydrocolloids,
2006, 20, 863-871.
[2] Naknean, P; Meenune M. Factors affecting retention and release of flavour compounds
in food carbohydrates. International Food Research Journal, 2010, 17, 23-34.
[3] Hodge, JE. Dehydrated Foods, Chemistry of Browning Reactions in Model Systems.
Journal of Agricultural and Food Chemistry, 1953, 1, 928-943.
[4] Namiki, M; Hayashi, T. Development of novel free radicals during the amino-carbonyl
reaction of sugars with amino acids. Journal of Agricultural and Food Chemistry,
1975, 23, 487-491.
[5] Arnoldi, A., et al., New Colored Compounds from the Maillard Reaction between
Xylose and Lysine. Journal of Agricultural and Food Chemistry, 1997, 45, 650-655.
[6] Hofmann, T. Characterization of the Chemical Structure of Novel Colored Maillard
Reaction Products from Furan-2-carboxaldehyde and Amino Acids. Journal of
Agricultural and Food Chemistry, 1998, 46, 932-940.
[7] Cämmerer, B; Jalyschko, W; Kroh LW. Intact Carbohydrate Structures as Part of the
Melanoidin Skeleton. Journal of Agricultural and Food Chemistry, 2002, 50, 2083-
2087.
[8] Franzke, C; Iwainsky, H. Antioxidant activity of melanoidins. Dtsch. Lebensm.
Rundsch., 1954, 50, 251-254.
[9] Elizalde, B; Rosa, M; Lerici, C. Effect of maillard reaction volatile products on lipid
oxidation. Journal of the American Oil Chemists' Society, 1991, 68, 758-762.
[10] Bedinghaus, AJ; Ockerman, HW. Antioxidative Maillard Reaction Products from
Reducing Sugars and Free Amino Acids in Cooked Ground Pork Patties. Journal of
Food Science, 1995. 60, 992-995.
[11] Yamaguchi, N. Antioxidant activity of the oxidation products prepared from
melanoidins. Developments in Food Science, 1986, 291-299.
[12] Yoshimura, Y., et al., Antioxidative Effect of Maillard Reaction Products Using
Glucos- lycine Model System. Journal of Agricultural and Food Chemistry, 1997, 45,
4106-4109.
104 Dongfeng Wang, Jipeng Sun, Guoqing Huang et al.

[13] Morales, FJ; Jiménez-Pérez, S. Free radical scavenging capacity of Maillard reaction
products as related to colour and fluorescence. Food Chemistry, 2001, 72, 119-125.
[14] Plakas, SM; et al., Effect of Maillard Browning Reaction on Protein Utilization and
Plasma Amino Acid Response by Rainbow Trout (Salmo gairdneri). The Journal of
Nutrition, 1985, 115, 1589-1599.
[15] Whitelaw, ML; Weaver, CM. Maillard Browning Effects on In Vitro Availability of
Zinc. Journal of Food Science, 1988, 53, 1508-1510.
[16] Nagao, M., et al., Mutagens in coffee and tea. Mutat Res, 1979, 68, 101-106.
[17] Martins, SIFS; Jongen, WMF; van Boekel, MAJS. A review of Maillard reaction in
food and implications to kinetic modelling. Trends in Food Science and Technology,
2000, 11, 364-373.
[18] Alink, GM; van Boekel, MAJS; Jongen, WMF. Mutagenicity of Heated Sugar-Casein
Systems: Effect of the Maillard Reaction. Journal of Agricultural and Food Chemistry,
2000. 48, 2271-2275.
[19] Monti, SM; Bailey, RG; Ames, JM. The influence of pH on the non-volatile reaction
products of aqueous Maillard model systems by HPLC with diode array detection.
Food Chemistry, 1998, 62, 369-375.
[20] Wedzicha, BL; Adamu, DJM. Reactivity of vegetables towards sulphur dioxide. Food
Chemistry, 1987, 26, 159-173.
[21] Kanner, J; Shapira, N. Oxygen- and Metal-Ion-Dependent Nonenzymatic Browning of
Grapefruit Juice, in Quality Factors of Fruits and Vegetables. American Chemical
Society, 1989, 55-64.
[22] Moreno, F.J., et al., High-Pressure Effects on Maillard Reaction between Glucose and
Lysine. Journal of Agricultural and Food Chemistry, 2002, 51, 394-400.
[23] Friedman, M., Food Browning and Its Prevention: An Overview. AU - Friedman,
Mendel. Journal of Agricultural and Food Chemistry, 1996, 44, 631-653.
[24] Chen, H., et al., Chemical composition analysis of soybean oligosaccharides and its
effect on ATPase activities in hyperlipidemic rats. International Journal of Biological
Macromolecules, 2010, 46, 229-231.
[25] Caicedo, L; Silva, E; Sánchez, O. Semibatch and continuous fructooligosaccharides
production by Aspergillus sp. N74 in a mechanically agitated airlift reactor. Journal of
Chemical Technology and Biotechnology, 2009, 84, 650-656.
[26] Lee, HW, et al., Chitosan oligosaccharides, dp 2-8, have prebiotic effect on the
Bifidobacterium bifidium and Lactobacillus sp. Anaerobe, 2002, 8, 319-324.
[27] Krastanov, A; Yoshida, T. Production of palatinose using Serratia plymuthica cells
immobilized in chitosan. Journal of Industrial Microbiology and Biotechnology, 2003,
30, 593-598.
[28] T., M., et al., Lactulose as a sugar with physiological significance. Bulletin 212,
International Dairy Federation, Brussels, 1987, 69-76.
[29] Galliard, T., Starch: Properties and Potential. 1st edition. Chichester: John Wiley and
Sons; 1987.
[30] Kamil, JYVA; Jeon, YJ; Shahidi,F. Antioxidative activity of chitosans of different
viscosity in cooked comminuted flesh of herring (Clupea harengus). Food Chemistry,
2002, 79, 69-77.
[31] Teleman, A., et al., Isolation and characterization of O-acetylated glucomannans from
aspen and birch wood. Carbohydrate Research, 2003, 338, 525-534.
Carbohydrates 105

[32] Huang, L., et al., Gelation Behavior of Native and Acetylated Konjac Glucomannan.
Biomacromolecules, 2002, 3, 1296-1303.
[33] Schwesinger, W.H., et al., Soluble dietary fiber protects against cholesterol gallstone
formation. The American Journal of Surgery, 1999, 177, 307-310.
[34] Yamamoto, K.;Yasuda, J; Too, K. Arrhythmias in newborn Thoroughbred foals.
Equine Veterinary Journal, 1992, 24, 169-173.
[35] Doi, K., et al., Influence of dietary fiber (konjac mannan) on absorption of vitamin B12
and vitamin E. Tohoku J Exp Med, 1983, 141, S677-S681.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 4

LIPIDS

Shengrong Shen1, Dongfeng Wang2 and Undurti N. Das3


1
College of Biosystem Engineering and Food Science, Zhejiang University, China
2
College of Food Science and Engineering, Ocean University of China, China
3
UND Life Science, USA Jawaharlal Nehru Technological University,
Kakinada, India

ABSTRACT
Lipids are a broad group of naturally-occurring molecules that are soluble in organic
solvents but insoluble or only sparingly soluble in water. Triacylglycerols account for
95% of total lipids in foods. Lipids are major energy source for human body and protect
human body from mechanical damage and heat loss. Lipids are also the carriers of fat-
soluble vitamins and the precursors of many bioactive molecules such as prostaglandin,
sex hormone, and epinephrine. This chapter deals with the nomenclature, classification,
and fatty acid distribution theories of lipids and their functional properties, including
polymorphism, plasticity, and emulsification. Lipids undergo multiple chemical reactions
during processing and storage, such as hydrolysis, oxidation, and thermal decomposition.
The effects of these reactions, especially oxidation, on food flavor and safety are
elucidated in detail. Lipid processing techniques, including refinement, hydrogenation
and interesterification, are also concerned in this chapter.

1. INTRODUCTION
It should be understood that though the term lipid is used majority of the times as a
synonym for fats, in fact, fats are a subgroup of lipids called triglycerides. Lipids encompass
molecules such as fatty acids and their derivatives including tri-, di-, and monoglycerides and
phospholipids, as well as other sterol-containing metabolites such as cholesterol. Humans and
other mammals have a variety of pathways to synthesize and metabolize lipids. Some lipids
cannot be synthesized by mammals and therefore must be ingested from the diet. Such lipids
are termed essential fatty acids (EFAs).
108 Shengrong Shen, Dongfeng Wang and Undurti N. Das

All plants and animals eaten by humans contain lipids. Various vegetables and most fruits
contain very little amounts of lipids, generally about 0.3%, except avocado, whose edible
portion contains about 20% lipids. The lipid content in the muscle tissue of lean beef, fish,
white poultry, and shell fish is about 2%, about 3.7% in cow‘s milk, about 2-4% in grains,
about 30% in fatty pork, 32% in an egg yolk, and about 35% in fillets of fatty fish. Oil-
bearing nuts and seeds contain from 20% fat in soybeans to 65% fat in walnuts.

1.1. Nomenclature

1.1.1. Fatty Acids


Fatty acids refer to any aliphatic monocarboxylic acids that are released by the hydrolysis
of naturally occurring lipids. More than 800 natural fatty acids have been identified, of which,
most contain a linear chain and even number of carbon atoms. Based on the presence or
absence of double bonds, lipids are divided into saturated and unsaturated fatty acids.
Palmitic acid and stearic acid are examples of saturated fatty acids. Unsaturated fatty acids
contain two or more double bonds. For instance, oleic acid has only one double bond, linoleic
acid has two double bonds, linolenic acid contains three double bonds, and arachidonic acid
contains up to four double bonds.
Fatty acids can be named by using one of the following four nomenclature systems.

Trivial nomenclature

Lipids are named according to their sources, such as palmitic acid, lauric acid, butyric
acid, stearic acid, and oleic acid.

Systematic nomenclature

Carbon atoms are counted from the carboxylic acid end.

Symbol nomenclature

Fatty acids can be represented by a simple numerical expression consisting of two terms
separated by a colon, with the first term depicting the number of carbon atoms and the second
the number of double bonds. For example, hexadecanoic acid,

CH3CH2CH2CH2CH2CH2CH2CH2CH2CH2CH2CH2CH2CH2CH2COOH

can be named as 16:0 and linoleic acid (9, 12-octadecadienoic acid) can be presented as 18:2.
This notation can be ambiguous, because some different fatty acids might have the same
numbers.

Omega-x or n-x nomenclature


Lipids 109

Unsaturated fatty acids can be named in the n-x or ω-x system. In this system, the
terminal methyl carbon is designated as ω or n and x represents the location of the first double
bond. Carbon atoms are counted from the terminal methyl carbon (namely ω or n) to the
carbonyl carbon. Because all adjacent double bonds in all natural polyenoic acids (containing
2 to 6 double bonds) are spaced by a methylene, the locations of all other bonds can be
determined if the first double bond is specified. For example, linoleic acid is presented as
18:2(ω-9). Because the first double locates in the 9th carbon, the second double occurs
between C12 and C13.
The geometric configuration of double bonds is usually designated by the use of cis and
trans, indicating whether the alkyl groups are on the same or opposite sides of the molecule.
Unsaturated fatty acids occur naturally in the cis form, but the trans form is more
thermodynamically stable.
If the four groups attached to the two carbon atoms of a double bond are different, the
configuration can be designated according to the Cahn-Ingold-Prelog rules. In this system,
each group is assigned a priority. If the two groups with higher priority (greater atom number)
locate in the same side of the double bond, the letter Z is used to designate the configuration.
If the two groups are on the opposite sides, the letter E is used.
Table 4-1 lists the systematic and common names of some fatty acids in naturally-
occurring lipids.

Table 4-1. Nomenclature of some common fatty acids

Abbreviation Systematic name Common name Symbol


4:0 Butanoic Butyric B
6:0 Hexanoic Caproic H
8:0 Octanoic Caprylic Oc
10:0 Decanoic Capric D
12:0 Dodecanoic Lauric La
14:0 Tetradecanoic Myristic M
16:0 Hexadecanoic Palmitic P
16:1(n-7) 9-Hexadecanoic Palmitoleic Po
18:0 Octadecanoic Stearic Sta
18:1(n-9) 9-Octadecanoic Oleic O
18:2(n-6) 9,12-Octadecanoic Linoleic L
18:3(n-3) 9,12,15- Octadecanoic Linolenic Ln
20:0 Arachidic Eicosanoic Ad
20:4(n-6) 5,8,11,14-Eicosatetraenoic Arachidonic An
20:5(n-3) 5,8,11,14,17-Eicosatetraenoic EPA
22:1(n-9) 13-Docosenoic Erucic E
22:5(n-3) 7,10,13,16,19-Docosapentaenoic
22:6 (n-6) 4,7,10,13,16,19-Docosahexaenoic DHA
Note: aSome authors use S for stearic, but this can be confusing, since S is also used for ―saturated‖
whenever triaclycerol composition is expressed in terms of saturated (S) and unsaturated (U) fatty
acids. For example, S3 or SSS=all three fatty acids saturated, SU2 or SUU = diunsaturated
monosaturated, and so on.
110 Shengrong Shen, Dongfeng Wang and Undurti N. Das

Figure 4-1. Structure of acylglycerols. R denotes fatty acid; there may be fatty acid, two fatty acids or
three fatty acids in the acylglycerols, which is mono-acyglycerol, di-acyglycerol or triacylglycerol
respectively; R1=R2=R3, the triacylglycerol is defined as simple triacylglycerol, otherwise mixed
triacylglycerol.

1.1.2. Acylglycerols
Lipids are the mixtures of mono-, di-, and triesters of glycerol with fatty acids, which are
termed monoacylglycerols, diacylglycerols, and triacylglycerols, respectively. Natural fats are
mainly present as triacylglycerols.
In order to designate the configuration of glycerol derivatives, the stereospecific
numbering (Sn) system is used in acylglycerol nomenclature. In this system, the carbon atom
that appears on top in that Fischer projection that shows a vertical carbon chain with the
hydroxyl group at carbon-2 to the left is designated as C-1 and the prefix 'sn' (for
stereospecifically numbered) is printed immediately preceding the glycerol term. An example
is shown in Figure 4-1. If R1 denotes stearic acid (St), R2 oleic acid (O), and R3 linoleic acid
(L) in the structure, this triacylglycerol is named Sn-glycerol-1-stearic acid-2-oleic acid-3-
linoleic acid, Sn-18:0-8:1-18:2, or Sn-StOL.
If the product is the racemic mixture of two antipodes, the prefix 'rac-' (for racemo) can
be used preceding the full name, such as in rac-StOM. In this case, the middle acid (O,
standing for the oleic acid) is in position 2 and the remaining two fatty acids (St and M,
standing for stearic and myristic acids respectively) are equally distributed in positions 1 and
3.
If the prefix β is used before the name, such as β-StOM, it indicates that the middle acid
is in position 2, but the distribution of the other two acids is unknown.
In the case of simple triacylglycerols or those with unknown fatty acids distribution, the
prefix can be omitted. For example, StOM can be used to express any mixture of Sn-StOM,
Sn-MOSt, Sn-OStM, Sn-MStO, Sn-StMO, and Sn-OMSt.

1.1.3. Phospholipids
Phospholipids refer to any lipids that contain phosphoric acid in the structures as mono-
or diester. According to the type of hydroxyl donors, phospholipids are divided into
phosphoglycerides and sphingolipids (sphingophospholipids), in the former structure the
alcohol is glycerol and the later sphingosine.

Figure 4-2. Structure of lecithin.


Lipids 111

In phospholipids, amino alcohols, such as choline, ethanolamine, serine, and inositol,


attach to phosphoric acid via an ester linkage, which are known as phosphatidylcholine,
phosphatidyethanolamine, phosphatidyserine and phosphatidylinositol respectively.
Phosphoglycerides are named as the derivatives of phosphatidic acids or similar to the Sn
system. For example, lecithin, whose structure is shown in Figure 4-2, is named as 3-sn-
phosphatidylcholine or sn-glycerol-1-stearic acid-2-linoleic acid-3-phosphocholine.
Sphingophospholipids adopt sphingosine as their backbone. The amino group in carbon 2
is connected to a long-chain fatty acid through the amido linkage to form ceramide and the
hydroxyl in carbon 3 is esterified by phosphoric acid. Choline or neovaricaine then attaches to
the phosphoric acid to produce various sphingomyelins.

1.2. Classification

Based on chemical structure and composition, lipids can be divided into simple lipids,
compound lipids, and derived lipids.
Simple lipids are compounds consisting of fatty acids and alcohols and include fats, oils,
and waxes.
Compound lipids contain many other groups in addition to fatty acids and alcohols.
Glycerophosphonolipids, sphingomyelins, cerebroside, and ganglioside are the examples of
compound lipids.
Derived lipids are the derivatives of simple and compound lipids and have the common
properties of lipids. Steroids, hydrocarbons, carotenoid, and fat-soluble vitamins are examples
of this group.
Most lipids in foods are acylglycerols. According to composition, lipids in animal and
plant-derived foods are divided into the following:

Milkfat

Milkfat exists in the latex of mammals. Milkfat consists mainly of palmitic acid, oleic
acid, and stearic acid. Compared with other animal-derived lipids, milkfat contain abundant
C4~C12 short-chain fatty acids, small amounts of odd- and branch-chain fatty acids, and
trans double bonds.

Laurates

Laurates are mainly found in palme plants such as coconut palm and babassu. Laurates
are featured by their high lauric acid contents, which reach up to 40%~50%. Laurates have
medium contents of C6, C8, and C10 fatty acids, low unsaturated fatty acid contents, and low
melting points.

Plant butter

Plant butter originates from the seeds of tropical plants and is characterized by narrow
melting point range. Although plant butter has a higher content of saturated fatty acids than
112 Shengrong Shen, Dongfeng Wang and Undurti N. Das

unsaturated fatty acids, trisaturated glycerides have not been identified in plant butter. Plant
butter is widely used in candies and cocoa butter is the most important plant butter.

Oleate-linoleate

Lipids of this category are the most abundant in nature and are found only in plants. The
lipids have high contents of oleic acid and linoleic acid and the contents of saturated fatty
acids are less than 20%. Peanut oil, corn oil, olive oil, palm oil, sesame oil, cottonseed oil,
and sunflower oil are important members of this category.

Linolenates

Linolenates contain large amount of linolenic acid and soybean oil, wheat germ oil, and
perilla oil belong to this category.

Animal fats

Lipids of this group, such as lard and tallow, are the storage fat of domestic animals.
Animal fats contain large amount of C16 and C18 fatty acids and medium contents of
unsaturated fatty acids, mostly oleic and linoleic acids, and small amount of odd-numbered
fatty acids. Animal fats contain appreciable amount of fully saturated triacylglycerols and
thus exhibit high melting points.

Marine oils

Marine oils contain abundant long-chain unsaturated fatty acids and are rich in vitamins
A and D. Eicose pentaenois acid (EPA) and docosahexenoic acid (DHA) are typical examples
of this category. Due to high unsaturation degree, marine oils are susceptible to oxidation.

1.3. Distribution of Fatty Acids in Nature Lipids

1.3.1. Theories of Triacylglycerol Distribution Pattern


In addition to fatty acid species and content, the properties of lipids are also affected by
the distribution of the fatty acids. Many theories on the fatty acid distribution of
triacyglycerols have been proposed and the most important are illustrated below:

Even or widest distribution theory

This theory was proposed by Hilditch and Williams in 1964 [1]. According to this theory,
the fatty acids in triacylglycerols tend to be distributed as broadly as possible. In this case,
when an acid, S, accounts for less than one-third of the total fatty acids present, it should not
appear more than once in any triacylglycerol. If X refers to the other fatty acids present, only
XXX and SXX species should be found. If a fatty acid accounts for between one-third and
two-thirds of the total fatty acids, it occurs at least once but never three times in any one
Lipids 113

molecule; that is, only SXX and SSX should be present. If a fatty acid forms more than two-
thirds of the total acids, it should occur at least twice in every molecule; that is, only SSX and
SSS should be present.
However, analysis of many natural fats, especially those of animal origin, reveals marked
deviation from this theory. Trisaturated acylglycerols have been found in fats containing less
than 67% saturated fatty acids. This theory is applicable only to triacyglycerols that consist of
two components and does not take positional isomers into consideration. This theory is no
longer considered valid.

Random (1, 2, 3-Random) distribution theory

According to this theory, fatty acids are distributed randomly both within each
triacylglycerol molecule and among all the triacylglycerols. Thus, the fatty acid composition
of all the three positions of glycerol should be the same and also equivalent to the fatty acid
composition of the total fat. The proportion of any given fatty acid species can be calculated
according to the following equation:

%sn-XYZ = (mol% X in total fat) × mol% Y in total fat) × mol% Z in total fat) × 10-4.

For example, if a fat contains four fatty acids, including 8% palmitic acid (P), 2% stearic
acid (St), 30% oleic acid (O), and 60% linoleic acid (L), 64 possible triacylglycerol species
would exist. The percentages of any species can be calculated by taking the following
equations as example:

Sn-OOO(%) = 30%×30% ×30% ×10-4 = 2.7.


Sn-PLSt(%) = 8×60×2×10-4 = 0.096.
Sn-LOL (%) = 60×30×60×10-4 = 10.8.

The fatty acid distribution of most lipids does not conform to the random distribution
theory. In natural lipids, the fatty acids in the Sn-2 position are different from those in
positions Sn-1 and Sn-3. Hence, great deviation exists between the predicted and actual
distribution of fatty acids in natural lipids. However, this theory can be used to predict the
fatty acid distribution in randomly interesterified fats.

Restricted random distribution theory

This theory was initiated by Kartha in 1953 [2]. According to this theory, saturated and
unsaturated fatty acids in animal fats are distributed randomly. Fully saturated triacylglycerols
(SSS) can be present, but only to the extent that the fat can remain fluid in vivo. Excess SSS,
according to this theory, can exchange with UUS and UUU to generate SSU and SUU.
Kartha's calculations do not account for positional isomers or positioning of individual acids.

1, 3-Random-2-Random distribution
114 Shengrong Shen, Dongfeng Wang and Undurti N. Das

This theory was proposed by Vander al, Coleman, and Fulton between 1960 and 1961
based on their researches of directed hydrolysis of Sn-1 and Sn-3 fatty acids by steapsin.
According to this theory, the fatty acid composition at position 2 is different and independent
from that of positions 1 and 3 and fatty acids are distributed randomly in Sn-2 and Sn-1,3.
Thus, the composition at positions 1 and 3 will, presumably, be identical. According to this
theory, the content of any given triacylglycerol can be calculated with the following equation:

Sn-XYZ(%) = (mol%X in positions 1 and 3)×(mol%Y in position 2)×(mol%Z in


positions 1 and 3)×10-4.

This theory is very useful in the exact predication of plant-derived lipids.

1-Random-2-Random-3-Random distribution theory

This theory was proposed by Tsuda in 1962. According to this theory, fatty acids are
separately but randomly distributed at each of the three positions of the triacylglycerol
molecules of a natural fat. Thus, the possibility of a given fatty acid appearing in each Sn
position would likely be different. The content of a given triacylglycerol species can be
calculated as:

%sn-XYZ = (mol% X at sn-1)×(mol% Y at sn-2)×(mol% Z at sn-3)×10-4.

This theory is applicable to the predication of common animal fat, milkfat, and seed fats.

1.3.2. Positional Distribution of Fatty Acids in Natural Fats


Generally, the Sn-2 position is preferred by unsaturated fatty acids, especially linoleic
acid, in seed-derived fats and saturated fatty acids are distributed at Sn1- and Sn-3 positions.
In most cases, the distributions of both saturated and unsaturated fatty acids at positions Sn-1
and Sn-3 are equivalent.
Animal-derived fats have higher saturated fatty acid contents at position Sn-1 than plant-
derived fats and their fatty acid compositions at positions Sn-1 and Sn-3 also differ markedly.
In most animal fats, palmitic acid is esterified at position Sn-1 with priority, and myristic acid
is esterified at position Sn-2.

Table 4-2. Positional distribution of fatty acids in triacylglycerols of some natural fats
[3]

Source Position 14:0 16:0 18:0 18:1 18:2 18:3 20:0 20:1 20:2 20:0
Coconut 1 34 50 12 1
2 2 2 87 9
3 37 53 9 -
Peanut 1 14 5 59 19 1 1 1
2 2 - 59 39 - - 0.5
3 11 5 57 10 4 3 6 3
Soybean 1 14 6 23 48 9
2 1 22 70 7
3 13 6 28 45 8
Lipids 115

Table 4-2. (Continued)

Source Position 14:0 16:0 18:0 18:1 18:2 18:3 20:0 20:1 20:2 20:0
Beef 1 4 41 17 20 4 1
2 9 17 9 41 5 1
3 1 22 24 37 5 1
Pig 1 1 10 30 51 6
2 4 72 2 13 3
3 - - 7 73 18

Lard has different fatty acid distribution patterns compared with other animal-derived
fats. For example, palmitic acid is mainly found in position Sn-2, stearic acid is distributed in
Sn-1, linoleic in Sn-3, and oleic acid in Sn-3 and Sn-1.
In milkfat, short-chain fatty acids attach specifically to position Sn-3, while the long-
chain polyunsaturated fatty acids are preferably esterified at position Sn-2 of marine oils.
The fatty acid distributions of some natural fats are listed in Table 4-2.

2. PHYSICAL PROPERTIES OF LIPIDS


Pure oils and fats are odorless and colorless, but processed fats and oils often display
yellow-green color due to the incomplete decoloration of such pigments as carotenoid and
chlorophyll. The odor of fats and oils is often attributed to the presence of non-lipid
impurities and lipid oxidation.

2.1. General Physical Properties

2.1.1. Density
The density of fatty acids and glycerides reduces with the increase of carbon chain length
and the decrease of degree of unsaturation. Generally, fatty acids and glycerides with high
hydroxyl and carbonyl contents have the highest density.
Natural lipids are the mixture of multiple acylglycerols and thus the relationship between
density and composition is rather complex. The densities of lipids are smaller than that of
water at room temperature and tend to decrease in higher temperatures. It is estimated that,
when a triacylglyceride is melted, its density decreases by about 10%.

2.1.2. Refractive Index


The refractive index of lipids rises with the increase of degree of unsaturation and chain
length. The refractive index of a monoacylglycerol is generally higher than that of the
corresponding triacylglycerol.
The refractive index is sometimes used in lipid identification to determine whether
adulteration occurs.
116 Shengrong Shen, Dongfeng Wang and Undurti N. Das

2.1.3. Melting Point


The melting point of saturated fatty acids is closely related to their chain length.
Generally, the index increases as the chain length rises. However, the melting points of odor-
numbered fatty acids are lower than that of the exactly adjacent even-numbered fatty acids
and the differences decrease as the chain length increases.
The melting points of unsaturated fatty acids are lower than their saturated forms and are
affected by the number, location, and configuration of double bonds. Fatty acids with more
double bonds have lower melting points and those with double bonds close to the two ends
exhibit higher melting points.
In the case of fatty acids with the same carbon number, branched ones are melted in
lower temperatures than those of straight chain ones and hydroxyl-containing ones have
higher melting points due to hydrogen bonding.
For acylglycerols, monoacylglycerols have the highest melting point, followed by
diacylglycerols and triacylglycerols in sequence. The melting point of triacylglycerols is
positively correlated with the content and chain length of fatty acids due to the increase of
inter-molecular action. The presence of trans fatty acids increases the melting point compared
with cis fatty acids, because the interaction between fatty acids is hindered in the cis
configuration. Similarly, because conjugation enhances the interaction between fatty acids,
lipids containing conjugated double bonds are melted at higher temperatures than those
without conjugated double bonds.
Natural lipids are the mixture of multiple acylglycerols and hence no specified melting
points can be given. The melting points of natural lipids are defined as a range of temperature
from lipid melting initiation to completion.

2.1.4. Smoke Point, Flash Point and Fire Point


The smoke point refers to the temperature at which a lipid begins to produce smoke,
which indicates the decomposition of the lipid. The smoke point of lipids is normally in the
range of 200–230 °C.
The flash point of a volatile liquid is the lowest temperature at which it can vaporize to
form an ignitable mixture in air.
The fire point, a higher temperature, is defined as the temperature at which the vapor
continues to burn for over 5 seconds after being ignited.
Smoke point, flash point, and fire point are three important properties that determine the
quality of lipids and can be used to determine the presence of impurities.

Figure 4-3. Crystalline forms of triacylglycerols.


Lipids 117

For example, the smoking point of refined edible oil is generally higher than 240 °C, but
the presence of free fatty acids dramatically decreases the index. The flash point of plant-
derived edible oil is generally over 225-240 °C and the fire point is usually 20-60 °C higher
than the flash point.

2.2. Polymorphism of Lipid

2.2.1. Crystallization and Polymorphism


Polymorphic forms are crystalline phases of the same chemical composition but differ in
structure and yield identical liquid phases upon melting. The polymorphic forms have
different stabilities and the less stable forms can automatically and unidirectionally transform
to the more stable forms without melting. If all the polymorphic forms are in their stable
states, the transformation is multidirectional and is decided by the temperature. When lipids
are placed in a temperature below freezing point, multiple crystal forms can be identified.
Polymorphism is an important property of lipids.
X-ray diffraction and infrared spectrum analysis indicate the existence of three crystal
forms, namely α, β’, and β in triacylglycerols, as shown in Figure 4-3. In the α form, the fatty
acid chains are arranged randomly. Hence, crystals in this form have lower melting point,
density and stability. Crystals in the β’ and β forms are arranged more orderly, especially for
the β form, in which the fatty acids are packed in the same direction. Hence, β’ and β form
crystals exhibit higher melting point, density, and stability.
The polymorphic behavior of natural lipids is affected by fatty acid composition and their
distribution in triacylglycerols. Generally, lipids consisting of relatively few closely related
triacylglycerol species tend to transform rapidly to the stable β form, while those consisting of
more triacylglycerol species are transformed slowly to the β' form. Soybean oil, peanut oil,
corn oil, olive oil, coconut oil, safflower oil, cocoa butter and lard are present mainly in the β
form, but cottonseed oil, palm oil, rapeseed oil, and milkfat are found mainly in the β' form.

2.2.2. Effects of Polymorphism on Food Processing


The different crystal forms in polymorphism have different physicochemical and sensory
properties. Conditions must be well controlled in food processing to yield desirable crystal
forms.
One example is the production of salad oil from cottonseed oil. Cottonseed oil must be
winterized to remove high-melting-point fats before further processing. During winterization,
the temperature must be decreased slowly to ensure sufficient time for bulk crystallization in
the β form, which is beneficial for subsequent filtration. If the temperature is reduced too fast,
fine crystals are formed and the filtration process becomes difficult.
Crystals in the β' form can impart desired coating performance and mouthful for
margarine. In margarine production, oils are firstly rapidly cooled to produce α form crystals
and then cooled in a slightly higher temperature to enable the transformation from α form to
the more stable β' form.
Chocolate should melt at 35 °C in the mouth without causing the greasy feeling. Besides,
chocolate must has smooth surface and the crystal particles cannot be too rough. For this
purpose, conditions must be controlled exactly to yield the desired β form. Cocoa butter is
118 Shengrong Shen, Dongfeng Wang and Undurti N. Das

heated to above 55 °C until completely melts and is then allowed to cool slowly until 29 °C.
The crystals are reheated to 32 °C to melt other crystal forms than the β form. The later
process is repeated until cocoa butter is completely transformed to the β form.

2.3. Plasticity

Lard and butter that are solids in room temperature are actually the mixtures of liquid oils
and solid fats. The lipids are completely solidified only in very low temperatures. Such
homogeneous mixtures of liquid oils and solid fats are defined as the plastic fat. Plastic fat
can maintain their shape even in exposure to external forces. The plasticity of fats is affected
by the following factors:

Fat crystal form

Fat crystals in the β' form have the best plasticity, because substantial amount of small
bubbles are trapped in the β' crystals. In contrast, only a small amount of air bubbles are
trapped in β crystals and the air bubbles are large in volume. Hence, fats in the β form have
less plasticity.

Melting point range

Fats with wider melting point ranges exhibit better plasticity.

Solid to liquid ratio

The fat/oil ratio significantly affects lipid plasticity. If the percentage of the solid phase is
too high, rigid crosslinking occurs and the fat exhibits high hardness and poor plasticity. If the
percentage of liquid phase is too high, the fat is flowable, soft, and susceptible to
deformation.

Figure 4-4. Heat content (H) or dilatometric (D) melting curve of a glyceride mixture [3].
Lipids 119

The solid phase to liquid phase ratio is also termed as the solid fat index (SFI). SFI is the
most important index that determines the plasticity of lipids and can be determined by
measuring the expansion of the fat, i.e. the volume increase of the fat during its transition
from solid to liquid.
The volumes of solid fat and liquid oil increase when heated. The expansion without
phase transformation is defined as thermal expansion. When solid fat is transformed to the
liquid state in high temperatures, the volume increase is called melting dilation. The melt-
expansion curve of plastic fats can be followed by measuring the specific volume change of
liquid oil and solid fat in different temperatures. As shown in Figure 4-4, solid fat starts to
melt at point X and is gradually melted until to point Y, at which the fat is completely
liquefied. The fat in point b is a mixture of solid and liquid. In this point, the percentage of
solid fat is ab/ac and that of liquid oil is bc/ac. Hence, the SFI in point b is ab/bc.
If a lipid melts in a very narrow temperature range, the slope between points X and Y is
high. Otherwise, the lipid exhibits a wide plastic range. Hence, the plastic range of lipids can
be modified by adding lipids with different melting points.
SFI can be measured exactly by using a dilatometer. However, measurement with
dilatometer is time consuming and is applicable only to fats with SFI less than 50%. Pulsed
nuclear magnetic resonance is the most common method for SFI determination. Because
ultrasonic travels faster in solid fat than in liquid oil, ultrasonic methods have been developed
for SFI measurement.

2.4. Lipid Emulsification and Emulsifier

2.4.1. Emulsion
Emulsions are dispersed system composed of two immiscible liquids. In an emulsion
system, one phase is dispersed in another phase in the form of liquid crystal or droplets with
diameter ranging from 0.1 μm to 50 μm and is thus designated as the inner phase or dispersed
phase and is surrounded by the outer phase or continuous phase. According to the natures of
the two phases, two emulsion systems, including oil-in-water (O/W) and water-in-oil
emulsions (W/O), have been identified. Milk is an example of the O/W emulsion and cream is
an example of the W/O emulsion.
The formation of small dispersed droplets increases the area of the interface between the
two phases and the interface area increases exponentially when the droplet diameter is
decreased. Due to the increased interface area, high energy is required to maintain positive
free energy for the interfaces. Hence, emulsions are unstable thermodynamic systems and
demulsification might occur under specific conditions. Emulsions can be destabilized in one
of the following ways:

Creaming

When the two phases differ in density, one phase migrates to the top or bottom of the
emulsion due to gravity. Large droplet size of the dispersed phase and density difference
accelerate the creaming process.
120 Shengrong Shen, Dongfeng Wang and Undurti N. Das

Flocculation

When the amount of static charge on the surface of dispersed droplets turns insufficient,
the repulsion between droplets is reduced. As a result, the droplets attach to each other and
flocculation occurs. The membranes of droplets remain intact after flocculation.

Coalescence

The membranes of dispersed droplets are disrupted and the drops move to each other,
leading to coalescence. Coalescence reduces the interface area of the dispersed phase and
planar interface might occur when coalescence is serious.

Coalescence can be prevented by adding emulsifiers. Emulsifiers are surfactants


containing both hydrophilic and lipophilic groups and are located in the oil/water interface.
Emulsifiers decrease the interfacial tension and thus reduce the energy required for emulsion
formation. Although surfactants reduce the interfacial tension, the interfacial free energy is
still positive and emulsions are still thermodynamically unstable.

2.4.2. Emulsifier
Emulsifiers are divided into anionic, cationic and nonionic types according to structure
and property. Based on origin, emulsifiers can be divided into natural and synthetic ones.
Based on functions, emulsifiers are divided into surfactants, viscosity reinforcing agents and
solid absorption agents. According to polarity, emulsifiers are divided into hydrophilic and
lipophilic ones.
Fatty monoglycerides and their derivatives (including glycerol monostearate and stearic
acid glyceride), sucrose fatty acid ester, sorbitan fatty acid ester and their derivatives (such as
sorbitan monooleate (Span 80) and polyoxyethylene sorbitan monostearate (Span 60)), and
phospholipids (such as modified soybean phospholipid), are widely used in the food industry
as emulsifiers.

3. CHEMICAL PROPERTIES OF LIPIDS


3.1. Hydrolysis

Lipids can be hydrolyzed by acid, alkali, heat, and enzymes to release free fatty acids in
the presence of water. Triacylglycerols are firstly hydrolyzed to diacylglycerols, followed by
monoacylglycerols and finally glycerol in sequence.
Free fatty acids are not found in living animals, but can be liberated by enzymatic
hydrolysis after animals are slaughtered. When mature oil seeds are harvested, lipid
hydrolysis occurs. Free fatty acids are less stable than the corresponding glycerides and are
more susceptible to oxidative rancidity. Hence, oils are neutralized by alkali in refinement to
reduce free fatty acid content and improve the quality and shelf life.
Lipids 121

When foods are fried, water might migrate into hot lipids, leading to lipid hydrolysis and
fatty acid release. Frying decreases the smoke point of lipids and deteriorate the quality of
foods.
Lipid hydrolysis is not preferred in most cases. However, moderate lipid hydrolysis can
create special flavors for some foods. For example, in cheese production, microbes and milk
lipases are added to produce desired flavor. In breads and yogurts, lipid hydrolysis also
contributes to flavor formation.

3.2. Oxidation

Lipid oxidation is a major cause of quality deterioration of lipid-containing foods. Lipid


oxidation generates various off-flavors and leads to rancid (oxidative rancidity), which render
these foods less acceptable. In addition, oxidation decreases the nutritional quality of foods
and produces potential toxicants. The prevention of lipid oxidation has been recognized as an
important issue of lipid chemistry.

3.2.1. Autoxidation
The autoxidation of lipids is a typical free radical chain reaction and the autoxidation rate
is markedly inhibited by chemicals which can interfere with free radical reactions. Light and
free radical-producing substances catalyze the reaction and the quantum yields exceed unity
when the oxidation reaction is initiated by light. Lipid autoxidation yields high levels of
hydroperoxides (ROOH) and the induction periods of pure substrates are longer than mixture
substrates.
The lipid autoxidation process is comprised of three steps: initiation, propagation, and
termination.

Initiation: RH→R• +H•


Propagation: R• + O2 → ROO•
ROO• + RH → ROOH + R•
Termination: R• + R• → R-R
R• + ROO• → R-O-O-R
ROO• + ROO• → R-O-O-R + O2

In initiation, the hydrogen atom on the α-carbon atom of the double bonds of unsaturated
fatty acids or their triacylglycerols (designated as RH) is detached by metal catalysis or
exposure to light or heat and a fatty acid radical known as the alkyl radical (R•) is produced.
R• then reacts with oxygen to produce peroxy radicals ROO•, which grabbles the hydrogen
atom from α-carbon atom of another molecules (RH) to yield hydroperoxides (ROOH) and
new free radicals R•. The reactions are repeated and lead to the propagation of lipid
autoxidation. Once these free radicals are combined to produce stable non-free radical
products, the chain reaction is terminated.
Initiation needs high activation energy and is therefore the limiting step of lipid
autoxidation.
122 Shengrong Shen, Dongfeng Wang and Undurti N. Das

3.2.2. Formation of Hydroperoxides


Hydroperoxides are the main product in the early phase of lipid autoxidation and their
structures vary with substrates (unsaturated fatty acids). After the hydrogen carbon is
detached from the α-methylene group neighboring a double bond, oxygen attacks the α
carbon atom connected the double bond, resulting in the formation of hydroperoxides. The
reaction is usually accompanied by a shift in position of double bonds.

Oleate

In autoxidation, the hydrogen atoms in C8 and C11 of oleates are detached and two
allylic radical intermediates are generated. Oxygen then attacks the end carbons of each
radical intermediates and an isomeric mixture of 8-, 9-, 10-, and 11-allylic hydroperoxides are
produced, as shown in Figure 4-5.
The percentages of 8- and 11-hydroperoxides are slightly higher than those of the 9- and
10-isomers in the mixture. When autoxidation occurs at 25 °C, the product mixture contains
equalmolar cis and trans 8- and 11-hydroperoxides and predominant percentages of trans 9-
and 10-isomers.

Linoleate

The hydrogen atoms attached to position 11 is highly active. Hence, only one radical
intermediate is produced upon autoxidation and an equalmolar mixture of conjugated 9- and
13-diene hydroperoxides is generated. The two hydroperoxide products undergo geometric
isomerization and hence are found in both the cis, trans and the trans, trans forms, as shown
in Figure 4-6.

Linolenate

Linolenate contains two 1, 4-pentadiene structures and their autoxidation products are a
mixture of 9-, 12-, 13-, and 16-hydroperoxides (Figure 4-7).

Figure 4-5. Autoxidation of oleate [4].


Lipids 123

Figure 4-6. Autoxidation of linoleates [5].

Figure 4-7. Autoxidation of linolenate [4].

OOH CHO
H2
R C C C CHO R C C C CHO CH2 RCHO
H H H H H
CHO

Figure 4-8. MDA formation by unsaturated aldehydes.

C5H11 O C5H11

3C5H11CHO
O O

C5H11
Figure 4-9. Tripentyltrioxane formation by polymerization of three hexanals.
124 Shengrong Shen, Dongfeng Wang and Undurti N. Das

Each of the four hydroperoxides has multiple geometric isomers with the conjugated
diene system in the cis, trans or the trans, trans configuration, but the unconjugated double
bond is always cis. Linolenate autoxidation results in more 9- and 16-hydroperoxides than the
12- and 13-isomers, because oxygen prefers C9 and C16 and the 12- and 13-hydroperoxides
are decomposed rapidly. The 12- and 13-hydroperoxides can form six-membered peroxide
hydroperoxide via 1,4-cyclization or prostaglandin-like endoperoxides via 1,3-cyclization.

3.2.3. Decomposition of Hydroperoxides


Hydroperoxides are extremely unstable and are decomposed immediately after they are
formed. In the initiation of autoxidation, the rate of hydroperoxides formation is higher than
decomposition, but the trend is reversed in subsequent steps.
The decomposition of hydroperoxide involves the production of alkoxy radical and its
further breakdown, with the products including aldehydes, ketones, alcohols, acids, epoxy
compounds, and hydrocarbons. The aldehydes and ketones include nonanal, 2-isodecenoic
aldehyde, 2-ecenal, hexanal, cis-4-heptenal 2,3-pentanedione, 2,4- glutaconaldehyde, 2,4-
decadiene, and 2,4,7-decatrienal. The resultant epoxy compounds are mainly the homologues
of furan. Unsaturated aldehydes can be further oxidized to malonaldehyde (MDA) (Figure 4-
8), which not only deteriorate the flavor of foods, but also impose safety concerns.
Saturated aldehyde products can be easily oxidized to corresponding acids or generate
new products through polymerization and condensation. For example, tripentyltrioxane can
be produced through the polymerization of three hexanal molecules (Figure 4-9).
The reactions involved in lipid autoxidation and decomposition are given in Figure 4-10..

3.2.4. Factors Influencing Lipid Autoxidation


The following factors influence the autoxidation of lipids:

Fatty acid composition

The quantity, location, and geometrical configuration of double bonds in fatty acid
components significantly affect lipid autoxidation. Lipids containing more double bonds are
oxidized faster than those with less double bonds. For example, the relative oxidation rates of
arachidonic acid, linolenic acid, linoleic acid, and oleic acid are 40, 20, 10, and 1 respectively.
The cis fatty acids are more susceptible to oxidation than the trans isomers. Conjugated
double bonds are oxidized more easily than unconjugated double bonds. Saturated fatty acids
are much more resistant to oxidation than unsaturated fatty acids. Compared with
triacylglycerols, free fatty acids are oxidized more rapidly and the autoxidation rate of lipids
increases when the free fatty acid content exceeds 0.5%. Random distribution of fatty acids in
lipids decreases the autoxidation rate.

Temperature

High temperature accelerates lipids autoxidation, though high temperature decreases the
solubility of oxygen. This is because the release of free fatty acids and the decomposition of
hydroperoxides are enhanced in high temperatures.
Lipids 125

Figure 4-10. Schemes of lipid autoxidation and decomposition [6].

Oxygen concentration

The autoxidation of lipids is not affected by oxygen level in the case of sufficient partial
pressure of oxygen. However, when the partial pressure of oxygen is low, the lipid
autoxidation rate is proportional to the partial pressure of oxygen.

Surface area

The lipid autoxidation rate is positively correlated with the area of the interface in contact
with air. In the case of a large surface area to volume ratio, the decrease of oxygen partial
pressure exerts no significant effect on oxidation rate. In O/W emulsions, the autoxidation
rate is determined by the diffusion of oxygen to the oil phase.
126 Shengrong Shen, Dongfeng Wang and Undurti N. Das

Water activity

Lipids are oxidized very rapidly in dried foods with aw less than 0.1. When aw is
increased, lipid autoxidation rate decreases dramatically. When the aw reaches 0.3, lipids are
oxidized in the minimum rate. This is because water reduces the catalytic activity of metals,
quenches free radicals, enhances non-enzymatic browning (producing antioxidant
compounds), and prevents the contact between foods and oxygen. When aw increases in the
range 0.3~0.7, the water solubility of oxygen and the fluidity of catalysts increase and more
reaction sites on substrates are exposed, leading to accelerated oxidation. When the aw further
increases, the lipid oxidation rate is reduced again due to diluted oxygen and catalyst
concentrations.

Auxiliary oxidants

Some divalent or polyvalent transit metals with appropriate redox potentials, such as Co,
Cu, Fe, Mn, and Ni, are effective auxiliary oxidants. These metals can shorten the initiation
period and accelerate lipid oxidation in concentrations as low as 0.1 mg/kg.

Light and radiation

Visible light, UV light and high-energy radiation can initiate the formation of free
radicals and decompose hydroperoxides, thus stimulating lipid oxidation. Hence, lipids or
lipid-containing foods should be stored in dark place or packed with opaque materials.
Special attention should be paid to this issue in the radiation sterilization of foods.

Antioxidants

Antioxidants can delay and retard the autoxidation of lipids. The effects of antioxidants
on lipid oxidation are detailed in Section 3.2.7.

3.2.5. Photosensitized Oxidation


Photosensitized oxidation, also referred to as photoxidation, is the reaction between the
double bonds of unsaturated fatty acids and singlet oxygen. Photosensitized oxidation can
initiate the autoxidation of lipids. Pigments in foods are natural photosencitizers. After
exposure to light, photosencitizers are excited to reactive molecules. Photosensitized
oxidation occurs in one of two ways. When photosencitizers are excited, they directly react
with lipids to form free radicals and initiate the autoxidation of lipids; or react with triplet
oxygen (3O2) to produce active excited singlet oxygen. The resultant excited oxygen directly
attacks the atoms on double bonds of unsaturated fatty acids and leads to the formation of
trans hydroperoxides. The number of hydroperoxide species is twice of the number of double
bonds. Figure 4-11 is the mechanism of the photosensitized oxidation of linoleic acid.
Because singlet oxygen is extremely active, the rate of photosensitized oxidation is 1000
times faster than the autoxidation of lipids. Besides, the hydroperoxide products can be
decomposed to free radicals to initiate autoxidation.
Lipids 127

Singlet oxygen does not show obvious specificity to substrate structure and can react with
any double bonds in unsaturated fatty acids. The rate of photosensitized oxidation is
proportional to the number of double bonds and is not affected by their locations. This
property is evidenced by the rate constants of the reactions of singlet oxygen with oleic acid,
linoleic acid, linolenic acid, and arachidonic acid, which are 0.74 × 105 mol-1 · S-1 · L, 1.3 ×
105 mol-1 · S-1 · L, 1.9 × 105 mol-1 · S-1 · L, and 2.4 × 105 mol-1 · S-1 · L respectively. In the
case of triplet oxygen, it is much less active and the rate constant of the reaction with linoleic
acid is only 89 mol-1 · S-1 · L, which is 1450 times lower than that of singlet oxygen. Hence,
the oxidation initiated by singlet oxygen predominates in the oxidation of unsaturated fatty
acids.

3.2.6. Enzymatic Oxidation


Lipoxidases catalyze the oxidation of lipids. Lipoxidases are widely present in organisms,
especially plants. Lipoxidases act specifically and attacks only polyunsaturated fatty acids
that contain the cis,cis-1,4-pentadiene structure, such as that in oleic acid, linoleic acid,
linolenic acid, and arachidonic acid. The mechanism of lipoxidase-catalyzed oxidation of
lipids is elucidated by taking linoleic acid as an example.
After the hydrogen of the ω-8 methylene is detached, a free radical is formed and then
isomerized to the trans configuration. As a result, the ω-6 and ω-10 hydroperoxides with
conjugated double bonds are produced, as shown in Figure 4-12.
13 9

HOO OOH HOO OOH


13 12 10 9

Figure 4-11. Mechanism of the photosensitized oxidation of linoleic acid.

Figure 4-12. Mechanism of the enzymatic oxidation of linoleic acid.


128 Shengrong Shen, Dongfeng Wang and Undurti N. Das

In animals, lipoxidases specifically oxidize arachidonic acid to produce prostaglandin,


prothrombin, and other active compounds. The generation of beany flavor of soybean
products has been attributed to the lipid oxidation catalyzed by lipoxidase.

3.2.7. Lipid Antioxidation and Antioxidants


Oxidation reduces the quality of lipids and deteriorates the quality of lipid-containing
foods. The prevention of lipid oxidation is a major concern of oil chemistry. The oxidation of
lipids can be delayed or retarded by both chemical and physical methods, but antioxidants are
the most effective and economic methods.

Antioxidant mechanism of antioxidants

Based on action mechanism, antioxidants are divided into free radical scavengers, singlet
oxygen quenchers, hydroperoxide decomposers, enzyme inhibitors, and antioxidant
synergists.
Free radical scavengers are either hydrogen or electron donors. Hydrogen donors, such as
phenolic antioxidants, can convert free radicals to their more stable forms by supplying an H·
radical. Because the resultant free radical cannot initiate a new free radical reaction, the chain
reaction is terminated as a result. Antioxidants of the electron donor type exert their functions
in the similar mechanism.
Singlet oxygen quenchers such as vitamin E react with singlet oxygen and convert it to
less active ground state oxygen and the quenchers themselves are transformed to the excited
state, which release the energy and return to the ground state immediately.
Hydroperoxide decomposers inhibit lipid oxidation by converting hydroperoxides
generated in chain reactions to non-active compounds.
Superoxide dismutases catalyze the formation of ground state oxygen and peroxide from
superoxides. The produced peroxide is then hydrolyzed to water and ground state oxygen by
catalase.
Antioxidant synergists are used together with antioxidants and can enhance the
antioxidant capability of antioxidants. Citric acid, fumaric acid, amino acids, vitamin C and
its esters, tartaric acid, and phospholipids are common antioxidant synergists. Synergists
reduce the catalytic activities of metal ions as well by complexing with them. Besides, some
synergists act by reducing the intermediate free radicals generated by antioxidants.

Common antioxidants

Antioxidants can be natural or synthetic compounds according to their origins. The


antioxidants that are approved in China include tocopherol, tea polyphenol, bamboo leaf
flavone, propyl gallate, vitamin C, butylated hydroxyarisol (BHA), 2,6-butylated
hydroxytoluence (BHT), and tertiary butylhydroquinone (TBHQ).
Lipids 129

3.3. Chemical Reactions in High Temperatures

Lipids might undergo oxidation, degradation, polymerization and condensation and


produce lower fatty acids, hydroxyl acids, esters, aldehydes, dimmers, and trimers in high
temperatures. These reactions deteriorate the quality of foods and lead to darkening, viscosity
increase, iodine value decrease, smoke point reduction, acid value elevation, and even irritant
odor. The changes occurred during frying are associated with fatty acid composition, food
composition, frying temperature, frying duration, and presence of metal ions. The thermal
decomposition and polymerization reactions of lipids are illustrated in Figure 4-13.

Figure 4-13. Thermal degradation and polymerization of lipids.

Figure 4-14. Non-oxidative thermal degradation of saturated fatty acids.

3.3.1. Thermal Degradation


Both saturated and unsaturated fatty acids undergo oxidative or non-oxidative thermal
degradation in high temperatures.
For saturated fatty acids, non-oxidative thermal degradation occurs only in very high
temperatures, as shown in Figure 4-14:
In the presence of oxygen, saturated fatty acids undergo oxidative-thermal degradation.
All the methylenes of fatty acids can be attacked by oxygen, but those at α, β, and γ carbons
are attached with priority. The degradation products, namely hydroperoxides, are further
decomposed to aldehydes, ketones, and low molecular weight hydrocarbons.
The non-oxidative thermal degradation of unsaturated fatty acids yields various dimmers
and some low-molecular weight compounds. The oxidative thermal decomposition of
unsaturated fatty acids occurs in the same pathway of lipid autoxidation in low temperatures,
but in much higher rates.
130 Shengrong Shen, Dongfeng Wang and Undurti N. Das

3.3.2. Thermal Polymerization


The thermal polymerization of lipids might be oxidative or non-oxidative reactions. Non-
oxidative thermal polymerization follows the Diels-Alder reaction. That is, a conjugated
diene reacts with a double bond to produce a cyclohexene.

R1 R1

R3 R3


R4 R4

R2 R2

Figure 4-15. Inter-molecular non-oxidative thermal polymerization of lipids.

Figure 4-16. Intra-molecular non-oxidative thermal polymerization of lipids.

Thermal polymerization occurs either between different molecules (Figure 4-15) or inside
the same molecule (Figure 4-16).
In oxidative thermal polymerization, homolysis occurs in the α-carbon of a double bond
and the resultant free radicals react with each other to produce non-cyclic dimmers or add to a
double bond to yield cyclic or non-cyclic compounds.
The thermal polymerization of lipids deteriorates the quality of foods and adversely
affects food nutrition and safety. However, these reactions are desired in some cases. For
example, thermal polymerization contributes to the formation of special flavors in fried foods.

3.4. Lipid Processing

3.4.1. Refining
Oils extracted from the tissues of plants and animals by compression, organic solvent
extraction, and cooking contain free fatty acids, phospholipids, sugars, proteins, water,
pigments, and many other impurities. These impurities adversely affect the color, flavor, and
stability of lipids and some compounds, such as aflatoxin and gossypol, threaten the health of
consumers. Hence, the refinement process is required to remove these impurities.

Degumming

Phospholipids are removed in this step. Phospholipids are soluble in oils with low water
content. When the water content increases, phospholipids are hydrated and precipitated from
Lipids 131

oil. In degumming, oils are added with 2%-3% water or vapor and then agitated. The water
phase is then separated by settling or centrifugation. Carbohydrates and proteins are also
removed in this step.

Neutralization

To remove free fatty acids and part phospholipids and pigments, NaOH solutions of
appropriate concentrations are added to heated oils. The resultant foots or soapstock is
separated and used for making soap.
Gossypol and aflatoxin are also removed or destroyed in neutralization.

Bleaching

Chlorophyll, lutein, carotene, and other pigments affect the appearance of oil and
decrease the stability against oxidation. These coloring materials can be removed by
absorbents, such as floridin, emathlite, and active carbon. Other impurities, such as
phospholipids, soaps, and some oxidation products are also absorbed by the absorbents. The
absorbents are then separated by filtration.

Deodorization

The off-flavors of lipids are caused by the volatile compounds generated in lipid
oxidation. These compounds can be removed by distillation in reduced pressure. Some
nonvolatile off-flavor compounds are also destroyed by water steam and are then distilled
away.

Table 4-3. Content changes of harmful substances in rapeseed oil during refining(μg/kg)

Compounds Crude oil Neutralization Bleaching Deodorization


Anthracene 10.1 5.8 4.0 0.4
Phenanthrene 100 68 42 15
1,2-benzanthracene 14 7.8 5.0 3.1
3,4-benzopyrene 2.5 1.8 1.0 0.9

The quality of oils is obviously improved after refining. Table 4-3 lists the removal of
various harmful substances in rapeseed oil during refining. It should be noted that some
useful components, such as fat-soluble vitamins and natural antioxidants are lost during
refining. Hence, additional antioxidants are often added to refined oils to improve the
antioxidant capacity.

3.4.2. Hydrogenation
Hydrogenation is the addition of hydrogen to the double bonds of unsaturated fatty acids.
Due to the insufficient supply of natural solid fat, liquid oils are hydrogenated to produce
semisolid fats. Hydrogenated fats have gained wide applications in the production of
margarine and shortening oil.
132 Shengrong Shen, Dongfeng Wang and Undurti N. Das

Oils can be either partially or completely hydrogenated. The products of fully


hydrogenated are used in soap production, and partially hydrogenated products are mainly
used in the food industry. Iodine values in the range 60~80 are preferred for partially
hydrogenated products, because the products exhibit desired melting point and solidity and
good thermal and oxidation stability.
The hydrogenation of oils is catalyzed mainly by nickel. In partial hydrogenation, the
reaction occurs in pressure 1.5~2.5 atm and temperature 125-190 °C. The degree of
hydrogenation can be measured according to the change of the refractive index. When the
desired hydrogenation degree is reached, the reaction can be stopped by cooling and
centrifugation. Oils must be refined before hydrogenation and the hydrogen used must be
anhydrous and free of sulfur, SO2, ammonia, or other impurities. In addition to nickel,
platinum, copper, and copper-chromium mixture are also used in oil hydrogenation as
catalysts. However, platinum catalyst is expensive and copper can cause toxicity and its
separation is difficult. Nickel is the most widely used catalyst.
The hydrogenation reaction comprises three steps. Firstly, a carbon-metal complex is
formed at an end of an olefin bond. This intermediate then reacts with the hydrogen attached
to catalyst to form an unstable half-hydrogenated state, in which only one olefin bond is
connected with the catalyst and thus can rotate freely. The half-hydrogenated compound
reacts with another hydrogen atom to yield a saturated product and separates from the
catalyst.
Non-naturally occurring fatty acids may be generated during hydrogenation. Besides,
some double bonds are translocated and isomerized during the process. Hence, partially
hydrogenated oils are a complex mixture of multiple fatty acids. The products of linolenic
acid hydrogenation are shown in Figure 4-17.

Figure 4-17. Hydrogenation products of linolenic acid [3].

Table 4-4. Effects of operating conditions on hydrogenation selectivity and rate

Operating parameter SR Trans fatty acids Rate


High temperature High High High
High temperature Low Low High
High catalyst concentration High High High
High-intensity agitation High High High

The selectivity of unsaturated fatty acid hydrogenation can be controlled by selecting


different hydrogenation conditions and catalysts. The selectivity of hydrogenation is
measured by the selectivity ratio (SR).
SR is defined by Albright and is the ratio (rate of hydrogenation of linoleic to oleic)/(rate
of hydrogenation of oleic to stearic).
Lipids 133

Reaction rate constants can be calculated from the starting and ending fatty acid
compositions and the hydrogenation time. Selective hydrogenation reduces the formation of
fully saturated glycerides, avoids excessive hardening of fats, and increases fat stability by
decreasing the content of linoleic acid. However, selective hydrogenation yields undesired
amount of trans isomers that cannot be absorbed by human body.
As shown in Table 4-4, both catalyst species and operating conditions affect the
hydrogenation selectivity.

3.4.3. Interesterification
The distribution pattern of fatty acids in lipids markedly affects the properties of lipids.
Because these properties affect the application of lipids in industries, interesterification has
been performed to improve lipid performance. Interesterification changes the distribution
pattern of fatty acid in triacylglycerols and is thus of industrial importance. Both intra-
(Figure 4-18) and inter-molecular interesterification (Figure 4-19) can occur, depending on
the composition and structure of triglycerides.
Interesterification can be accomplished by heating lipids in high temperatures for a long
period. In the presence of catalyst, interesterification occurs at lower temperatures in a short
period (50°C, 30min). Alkali metals and alkylated alkali metal are effective low-temperature
catalysts and sodium methoxide is the most common one. The dosage of catalyst is generally
about 0.1% of lipids.

R1 R2 R1

R2 R1 R3

R3 R3 R2

Figure 4-18. Intra-molecular interesterification.

R1 R2 R1 R2
R1 + R2 R2 + R1

R1 R2 R1 R2

R1 R2

R1 + R2

R2 R1

Figure 4-19. Intermolecular interesterification.

Excessive catalyst leads to more lipid loss due to the formation of soap and methyl esters.
Impurities, such as free fatty acids and peroxides, can react with sodium methoxide and
these impurities must be present in low levels in interesterification. Besides, sodium
methoxide can react with water and lost the catalytic activity. Hence, interesterification
reactions can be terminated by adding water.
134 Shengrong Shen, Dongfeng Wang and Undurti N. Das

Figure 4-20. Effect of interesterification on solid content index of lard [7].

Interesterification occurs either in the directed or the randomized mode. If the


temperature is higher than the melting point of lipids, randomized interesterification occurs.
In this mode, fatty acids are randomly connected to glycerol and the proportions of various
glycerides depend on the contents of fatty acid contents in original lipids, which can be
calculated according to the 1, 2, 3-random distribution theory mentioned previously. If the
temperature is lower than the melting point, directed interesterification occurs. In this case,
triglycerides with melting points higher than the reaction temperature crystallize and separate
from the reaction mixture and the reaction proceeds until most of the fatty acids are
precipitated. In directed interesterification, the proportions of trisaturated acylglycerols S3 (as
crystals) and triunsatured acylglycerols U3 (liquid) increase simultaneously.
Interesterification has found wide application in the manufacture of shortenings. Lard,
due to its high proportion of disaturated triacylglycerols with palmitic acid in the 2 position,
forms relatively large and coarse crystals, even when rapidly solidified in commercial chilling
machines. Randomized interesterification of lard improves its plastic range and makes it a
better shortening. Directed interesterification, however, produces a product with a higher
solid content at high temperatures and thus extends its plastic range, as shown in Figure 4-20.
Enzymatic interesterification utilizes lipase as catalyst. Compared with chemical
interesterification, enzymatic interesterification occurs in mild conditions with high
specificity, less byproducts, and low energy consumption. Hence, enzymatic
interesterification is a promising method for lipid processing. Enzymatic interesterification
has been used in the production of high-quality products such as cocoa butter equivalent and
breast milk fat substitutes.
Lipids 135

REFERENCES
[1] Hilditch, TP; Williams, PN. The Chemical Constitution of Natural Fats, 4th edition.
London: Chapman and Hall, 1964.
[2] Kartha, ARS. The glyceride structure of natural fats. II. The rule of glyceride type
distribution of natural fats. Journal of the American Oil Chemists' Society, 1953, 30,
326–329.
[3] Fennema, OR. Food Chemistry. 3rd edition. New York: Marcel Dekker; 1996.
[4] Chan, HW; Levett, G. Autoxidation of methyl linoleate: Analysis of methyl
hydroxylinoleate isomers by high performance liquid chromatography. Lipids, 1977,
12, 837-840.
[5] Chan, HW; Levett, G. Autoxidation of methyl linoleate: Separation and analysis of
isomeric mixtures of methyl linoleate hydroperoxides and methyl hydroxylinoleates.
Lipids, 1977, 12: 99-104.
[6] Frankel, EN. Autoxidation in Fatty Acids. Journal of the American Oil Chemists'
Society, 1979, 353-390.
[7] Sreenivasan, B. Interesterification of fats. Journal of the American Oil Chemists'
Society, 1978, 796–805.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 5

PROTEINS

Hong Lin1, Lisha Wu1 and Shuhui Wang2


1
College of Food Science and Engineering, Ocean University of
China, Qingdao, China
2
Biosystems Engineering Department, College of Agriculture – Ginn
College of Engineering, Aubum University, Auburn, AL, US

ABSTRACT
Protein is one of the most important components in foods. Most proteins are
constituted with 20 kinds of amino acids that are linked through peptide linkages. The
combination of these amino acids leads to the occurrence of substantial amount of
proteins with different structures and functions. Theoretically, all proteins produced by
organisms can be utilized by human body. However, only proteins that are readily
digestible, non-toxic and nutritious with specific functions are used in diets. This chapter
briefly describes the properties and structures of amino acid components and proteins and
highlights the properties of important dietary protein sources. Denaturation is an
important change in food processing. The mechanism of denaturation, properties of
denatured proteins and factors causing denaturation are also included in this chapter. In
addition to the nutrition attribute, some functional properties, including emulsifying,
foaming, gelling, and viscosity, also contribute to its wide applications in the food
industry. These functional properties are described in detail in this chapter. Proteins
undergo various changes during processing. These changes might greatly affect the
quality of foods. This chapter also emphasize on the changes of proteins occurred in
various processing techniques.
138 Hong Lin, Lisha Wu and Shuhui Wang

1. PHYSICAL AND CHEMICAL PROPERTIES OF


AMINO ACIDS AND PROTEINS
1.1. Amino Acids

1.1.1. Acid-Base Properties


Amino acids are amphiphilic molecules that contain an acidic carboxyl group and one
alkaline amine group. The dissociation constants of the amine and carboxyl groups in amino
acids are lower than common carboxyl and amino groups. For example, the Ka value of
glycine is 1.6×10-10, which is much lower than 10-5 of most carboxylic acids. Hence, the two
groups do not occur as free forms in most cases. At around neutral pH, amino acids are
present as dipolar ions or zwitterions.
Amino acids can react with both acids and bases. Carboxylic acid groups (–COOH) can
be deprotonated to become negative carboxylates (–COO-), and α-amino groups (–NH2-)
can be protonated to become positive α-ammonium groups (+NH3-), depending on the pH
of the environment. At pH values greater than the pKa of the carboxylic acid group, the
negative carboxylate ion predominates. At the pH values lower than the pKa of the α-
ammonium group, the nitrogen is predominantly protonated as a positively charged α-
ammonium group. Thus, at pH between the pKa of carboxylic and amine groups, amino acids
have net zero charge and this state is known as zwitterion. Below the pKa of carboxylic acid,
the predominant form will have a neutral carboxylic acid group and a positive α-ammonium
ion, and above the pKa of the amine group, most amino acids have a neutral amine group and
a negative carboxylate.

1.1.2. Isoelectric Point


At pH values between the two pKa values, the zwitterion predominates, but coexists in
dynamic equilibrium with small amounts of net negative and net positive ions. At the exact
midpoint between the two pKa values, the trace amount of net negative and trace of net
positive ions exactly balance, so that average net charge of all forms present is zero. This pH
is known as the isoelectric point pI, so pI = ½(pKa1 + pKa2). Amino acids have zero mobility
in electrophoresis and minimum solubility at their isoelectric points. Hence, amino acids can
be isolated by precipitation from water by adjusting the pH to their isoelectric points.
The pI ranges of neutral, acidic, and alkaline amino acids are 5.5~6.3, 2.8~3.2, and
7.6~10.6 respectively. Generally, amino acids with more amine groups have higher pI values
and those with more carboxylic groups have lower pI values.

1.1.3. Chemical Reactions


Amino acids show the usual reactions of both carboxylic acids and amines and these
reactions might occur during food processing.

(1) Acylation of amine group


Activated acid derivatives, such as acid halogenides and anhydrides, can react with the
amine group of amine acids as acylating agents. The following is the reaction between
carbobenzoxy chloride and amino acids
Proteins 139

The acylated products are more stable against various reagents and can be removed by
many methods. Hence, carbobenzoxy chloride has been widely used in protein synthesis for
amine group protection.

(2) Alkylation of amine group


Amino acids can react with 1-fluoro-2, 4-dinitrobenzene (FDNB) to yield N-2,4-
dinitrophenyl amino acids (DNP-amino acids), which are yellow compounds and crystallize
readily. This reaction is very important in N-terminal acid labeling for protein sequencing in
Sanger‘s method.

(3) Reaction with nitrous acid


All α-amino acids, except proline, can react with nitrous acid to yield nitrogen and
hydroxyl acid. The liberated nitrogen gas is contributed by equal molar amine group and
nitrous group. Hence, the contents of free amine groups in amino acids can be determined by
measuring the volume of nitrogen gas formed by reaction with nitrous acid. This is the
principle of the Van Slyke method.

(4) Reaction with ninhydrin


All α-amino acids, except proline, can react with ninhydrin in alkaline solutions to
produce purple products. This reaction has been widely used in the identification and
colorimetric quantification of amino acids.

(5) Reactions of carboxylic groups.


The carboxylic groups of amino acids can undergo various chemical reactions. Azide
reaction is a typical reaction on the carboxylic group. Amino acid ester reacts with hydrazine
to yield hydrazide and the product then reacts with nitrous acid to produce azide:

The azide compound can react with another amino acid ester by condensation to produce
a dipeptide:
140 Hong Lin, Lisha Wu and Shuhui Wang

Peptides synthesized in this method are optically pure.


When heated, the carboxylic group can react with the amine group of in the same
molecule to produce diketopiperazine. This compound contributes to the special flavor of
foods.

1.3. Sensory Properties

1.3.1. Bitterness
Amino acids contain multiple functional groups and can act with various taste receptors
to exhibit different tastes. Sweet amino acids are primarily found among amino acids of the D
form, while the tastes of L-series amino acids are significantly affected by their side chains.
When the side chain is small, the sweet taste predominates. For example, glycine (Gly),
alanine (Ala), proline (Pro), hydroxyproline (Hpr), homocysteine (Hcys), serine (Ser),
threonine (Thr), asparagine (Asn), and glutamic acid methyl ester (Glu-OMe) contain small
side chains and taste sweet. When the side chain contains more than three carbons, such as
leucine (Leu), isoleucine (Ile), norleucine (Nle), phenylalanine (Phe), tyrosine (Tyr),
tryptophane (Trp), histone (His), lysine (Lys), arginine (Arg), the bitter taste predominates. If
the side chain is of moderate size, the amino acids taste both sweet and sweet, such as valine
(Val) and ornithine (Orn). The sensory property of amino acids also depends on the
hydrophobicity of side chains. If the side chain is only slightly hydrophobic, the amino acid
tastes slightly sweet without bitterness, such as glutamin (Gln), cysteine (Cys), and
methionine. If the side chain is acidic, the amino acids taste sour, such as aspartic acid (Asp)
and glutamic acid (Glu). Table 5-1 lists the relationship between the sensory properties and
side chain structure of amino acids.

Table 5-1. Relationship between the side chain structure and taste of amino acids

Category Amino acids Side chain property Taste


І Glu, Asp, Gln, Asn Acidic Sour
II Thr, Ser, Ala, Gly, Met, Short Sweet
Cys
III Hpr, Pro Pyrrole Sweet, with slight
bitterness
IV Val, Leu, Ile, Phe, Tyr, Large and long Bitter
Trp
V His, Lys, Arg Alkaline Sweet, with slight
bitterness

The sensory properties of oligopeptides, especially dipeptides, are decided by the


original tastes of the amino acid components. The rules are as follows:
Proteins 141

(1) Neutral peptides consisting of type I and V amino acids and those containing only
type II amino acids exhibit light tastes.
(2) The sodium salts of oligopeptides consisting of only type I amino acids or type I and
type II amino acids taste fresh, such as Glu-Glu, Glu-Asp, Glu-Ser, and Glu-Thr.
(3) The combination of type III amino acids with type I amino acids remove the bitter
taste, but remain the sour taste.
(4) Peptides composed of type III, IV, or V amino acids only or their combinations are
bitter.
(5) Peptide formation, carboxyl group esterification, or diketopiperazine formation by
coupling of type IV and V amino acids enhances the bitter taste.
(6) Peptides with type IV and V amino acids locating at the C terminal are 3~5 times
bitterer than those with the amino acids locating at the N terminal or the middle of the
peptides.
(7) Type II amino acids (especially Gly) locating at either terminals or cyclized increase
the bitterness.

All peptides contain AH polar groups. However, because peptides differ markedly in
molecular weight and nature of hydrophobic groups, they have different bitterness receptor
binding capabilities. The bitterness intensity of peptides can be predicated by calculating the
average hydrophobicity of side chains, which is related to the total hydrophobicity of
nonpolar amino acids. Table 5-2 lists the hydrophobicity of various amino acids. The average
hydrophobicity of a peptide can be calculated by using the following formula:

In which, ⊿G is the free energy change of the side chains of amino acids and n is the
number of resides in the peptide.
Peptides with Q greater than 6.85kJ/mol are bitter and those with Q less than 5.43kJ/mol
are not bitter. No rules have been found for the taste of peptides with Q in the range
5.43~6.85kJ/mol. However, exceptions have been found. For example, all peptides with
molecular weight more than 6000 taste light despite of the Q value. If Gly is attached to one
or both ends of a peptide, the peptide tastes bitter even though Q is less than 5.43kJ/mol. This
is possible due to the less steric hindrance of Gly that facilitates the contact with bitterness
receptors. For this reason, it has been proposed to exclude Gly from Q calculation. Arg-Arg,
Lys-Ala, Val-Ah, Arg-Gly-Pro, and Ser-Lys-Gly-Leu, are also exceptions to the rule. The Q
values of these peptides are less than 5.43kJ/mol, but they taste bitter.
Besides, the bitterness intensity of peptides with similar Q values might differ
significantly. The bitterness intensity of Gly-Gly-Gly-Leu is greater than its three positional
isomers and that of Leu polymers are Leu-Leu-Leu-Leu, Leu-Leu-Leu, Leu-Leu, and Leu in
decreasing order. In the case of bitter peptides in cheese, the bitter taste disappears after the
Gln at the N terminal is cyclized, though the Q value is not changed. These inconsistencies
indicate that the bitterness intensity of peptides is also affected by their molecular weights and
higher structures in addition to Q values.
142 Hong Lin, Lisha Wu and Shuhui Wang

Table 5-2. Hydrophobicity of the side chain of amino acids


(transfer from ethanol to water)

Amino acid ⊿G0(side chain)/(kj/mol) Amino acid ⊿G0(side chain)/(kj/mol)


Ala 2.09 Leu 9.61
Arg 2.1 Lys 6.25
Asn 0 Met 5.43
Asp 2.09 Phe 10.45
Cys 4.18 Pro 10.87
Gln -0.42 Ser -1.25
Glu 2.09 Thr 1.67
Gly 0 Trp 14.21
His 2.09 Tyr 9.61
Ile 12.54 Val 6.27

1.3.2. Delicious Flavor


The delicious flavor is a complex taste and is generally contributed by various flavor
enhancers, including some amino acids, as shown in Table 5-3.
Table 5-3. Main delicious flavor-contributing components in some foods

Material MSG Amino acid amine 5‘-IMP 5‘-GMP Sodium


and peptides succinate
Animal meat + ++ +++
Fish meat + ++ +++
Shrimp, crab + + ++
Shellfish +++ +++ +++
Octopus, squid ++ +++
Kelp +++ ++
Vegetable ++
Mushroom +++
Soy sauce +++ +++
Note: MSG, monosodium glutamate; 5‘-IMP, 5‘-inosine monophosphate; 5‘-GMP, 5‘-guanosine monophosphate.

(1) Common properties of flavor enhancers.


Flavor enhancers are compounds that enhance the aroma of a food, though they
themselves have no distinct odor or taste in concentrations used. Many compounds have been
found to have the flavor enhancing capability.

Only dissociated glutamic acid salts can enhance flavor, of which, monosodium
glutamate (MSG) has the purest taste. Glutamic acid derivates that cannot dissociate
have no the flavor enhancing capability.
5‘-Inosine monophosphate (5‘-IMP), 5‘-guanosine monophosphate (5‘-GMP), and
5‘-xanthine monophosphate (5‘-XMP) are good flavor enhancers, but adenylate has
no the flavor enhancing ability.
L-Cys, sodium salt of thiosulfonic acid, homocysteine, L-Asp, L-aminoadipic acid,
and succinic acid, display flavor enhancing abilities similar to L-MSG.
Proteins 143

Many fruit acids, such as malic acid, tartaric acid, and citric acid, enhance the taste of
foods. The combination of anyone of the acids with lactic acid can improve the odor
of soybean product. Besides, citric acid juice enhances the taste of strawberry.
Fumaric acid and maleic acid can suppress the unpleasant odor of garlic.
Glutathione enhances the taste of meat broth. Polyphosphates improves the taste of
chicken meat and cheese.
The diamine salts of dicarboxylic acids with carbon atom number ranging from 3 to
10 can be used as the substitutes of table salt.

It is proposed that all flavor enhancers show a common structure O-(C)n-O, in which, n
ranges from 3 to 9. In another word, the compounds must contain an alkyl chain with 3~9
carbons and the two ends must be negative charged. Compounds with 4~6 carbons have the
highest enhancing ability. The alkyl chain can be straight, branched, or cyclic and the carbon
atoms can be replaced by O, N, S, and P. The negative charges on the two ends of the
molecule are crucial for the enhancing ability. If the carboxylic acid group is esterified,
amidated, or converted to lactone or lactam upon heating, the enhancing ability is obviously
reduced. The negative charge of an end can be replaced by a negative dipole. For example,
the enhancing abilities of tricholomic acid and ibotenic acid are 5~30 times higher than that
of L-MSG.

For economic and safety reasons, only Glu- and nucleotide-type flavor enhancers have
gained commercial applications.

(2) Glu-type flavor enhancers.


Flavor enhancers containing Glu are fatty compounds and have specific spatial
configuration for their functions. The negatively-charged groups at the two ends, such as –
COOH, –SO3H, and –SH, determine the flavor enhancing capability and hydrophilic groups,
such as –L-NH2OH, assist in the capability. Dipeptides or tripeptides with hydrophilic amino
acids attached to the carboxylic end of Glu have the flavor enhancing ability, but those with
hydrophobic amino acids attached are bitter, as shown in Table 5-4.
It is noteworthy that all the amino acids have more than one taste. Table 5-5 lists the
tastes of three amino acids.
The flavor enhancing capability of MSG is affected by the environmental pH. MSG has
the highest enhancing ability in pH6.0. Besides, temperature also affects the ability. When
MSG is heated for a long time or at 120°C, inter-molecular dehydration occurs and
pyroglutamic acid (shown below) is generated. This product losses the flavor enhancing
ability and is toxic to human body.
144 Hong Lin, Lisha Wu and Shuhui Wang

Table 5-4. Relative flavor enhancing abilities of MSG-containing flavor enhancers


(with L-MSG as 1)

Flavor enhancer Relatively ability


L-MSG 1.00
sodium salt of L-Asp 0.08
Sodium salt of L-α- aminoadipic acid 0.10
Sodium L- tricholomic acid 5~30
sodium salt of L-cysteine thiosulfonic acid 0.10
Glu-γ-NH2 Delicious
Glu-γ-Ome Sweet
Glu-α-NH2 0
Glu-Asp 0.15
Glu-Ser 0.10
Thr-Glu 0.10
Asp-Glu-Ser 0.10
Ser-Glu-Glu 0.15
Glu-Glu-Glu 0.20
Glu-Gly-Ser 2.00
Glu-Glu-Ser 2.00
Glu-α,γ-(NH2)2 Bitter
(Erythrose)-HO2CCH2CHOHCHNH2CO2Na 0.1
(Stachyose)- HO2CCH2CHOHCHNH2CO2Na 1.8
(R,S)-HO2CCHMeCH2CHNH2CO2Na 0.1
(S,S)-HO2CCHMeCH2CHNH2CO2Na 0.1
L-HO2CCHOHCHNH2CO2Na Nearly 0
L-NaO3SCH2CHNH2CO2Na 0.1
L-NaO3S(CH2)2CHNH2CO2Na 1.0
L-NaO3S(CH2)3CHNH2CO2Na 0.1
Ac-Glu-α,γ-(OMe)2 0

Table 5-5. Taste proportions of three amino acids

Amino acid Delicious taste Sour Salty Sweet Bitter


L-Glu 21.5% 64.2% 2.2% 0.8% 5.0%
L-MSG 71.4% 3.4% 13.5% 9.8% 1.7%
L-Try 1.2% 5.6% 0.6% 1.4% 87.6%

In addition, MSG is racemized under alkaline solutions when heated and loses flavor
enhancing ability. Hence, MSG should be added after dishes or soups are cooked.

(3) Nucleotide-type flavor enhancers.


Flavor enhancers of this type are aromatic heterocyclic compounds. The hydrophilic
ribose phosphate determines the flavor enhancing ability and e hydrophobic substituents
attached to the aromatic heterocycle assist in the ability.
Proteins 145

2. STRUCTURE OF PROTEIN
2.1. Primary Structure

Primary structure is the sequence of amino acid residues in proteins and is decided by
the genetic codes in genes. The primary structure decides the higher structures of proteins. All
proteins are composed of 20 amino acids and the combination of these components leads to
the occurrence of tremendous number of proteins with different spatial structures and
biological functions.

2.2. Higher Structures

The polypeptide chains of proteins occur as folded and twisted conformation under
suitable conditions and these conformations determine the biological and physicochemical
properties of the molecules in addition to their amino acid sequences.

2.2.1. Secondary Structure


Secondary structure is the three-dimensional form of local segments of polypeptides.
This term does not involve the conformation of the side chains of amino acid residues. Two
forms of periodic secondary structures have been found: α-helices and β-sheet.
The α-helical structure is characterized by the following:

(1) Multiple peptide bond planes rotate through the α-carbon atom spin, twisting each
other into a tight solid right-handed helix.
(2) Each helical rotation involves 3.6 amino acid residues. Each residue extends the axial
length by 1.5 Å and the angle of rotation per residue is 100°.
(3) α-Helices are stabilized by hydrogen bonding. In this structure, each backbone N-H
group is hydrogen bonded to the C=O group of the fourth preceding residue in the same
chain.
(4) The side chains of amino acid residues are located on the outer of the helix and their
shape, size, and charge affect helix formation. α-Helix is often not found in areas with
abundant acidic or basic amino acids due to charge repulsion or areas consisting more amino
acids with large side chains, such as tryptophan and isoleucin. Proline is generally not
146 Hong Lin, Lisha Wu and Shuhui Wang

involved in α-helix formation, because it cannot rotate or hydrogen bond with other amino
acids readily. The presence of glycine affects helix stability, because its side chain (H)
occupies the minimum space.

The β-sheet structure is characterized by the following:

(1) The β-sheet structure is an extended structure. Amino acid residues involved are
arranged in a zigzag pattern and their residues are located on the upside or downside of the
pleat plane. The angle between adjacent peptide bond planes is 110°.
(2) The conformation is stabilized by hydrogen bonding of C=O and –NH between
adjacent strands of a peptide.
(3) When adjacent strands run in the same direction, the peptide chains are parallel.
When the chains run in opposite directions, a planar, antiparallel sheet structure is stabilized.
Both parallel and antiparallel sheets can occur in one peptide.
(4) In parallel β-sheet, the distance between any two adjacent residues is 0.65 nm and
that in anti-parallel β-sheet is 0.7 nm.

2.2.2. Tertiary Structure


The secondary structures of a polypeptide chain can further fold to form a compact
three-dimensional structure termed as tertiary structure. The tertiary structure of proteins are
stabilized mainly by secondary bonds, including hydrogen bond, hydrophobic interaction, salt
linkage, and Vander Wasls force, as shown in Figure 5-1. These secondary bonds might occur
between the side chains of amino acid residues that are quite far away in primary structure.
Secondary bonds are non covalent bonds and are affected by the pH, temperature, and ionic
strength of the environment and are hence subject to change. Disulfide bond is not a
secondary bond, but it can connect two segments of a peptide that are far away in a peptide
chain. Hence, disulfide bonds are very important for stabilizing tertiary structures.
Proteins can be divided into two large groups on the basis of conformation: fibrous and
globular proteins. Fibrous proteins, such as fibroinare, the slender molecules with axial ratio
are greater than 10. Plasma albumin, globulin, and myoglobin, are examples of globular
proteins. In globular proteins, hydrophobic groups are located on the interior of a protein and
the hydrophilic groups are distributed on the surface. Hence, globular proteins are hydrophilic
molecules and most enzymes are globular proteins. The distribution of the side chains leads to
the formation of special areas with special biological functions, such as the active center of
enzymes. Figure 5-2 shows the folding patterns of two proteins.

2.2.3. Quaternary Structure


Multiple folded peptide chains can combine with each other through secondary bonding
to form a more complex structure. This structure is called the quaternary structure of proteins.
Each peptide chain with specific tertiary structures is a subunit of the protein. Hence,
quaternary structure is also defined as the arrangement and interaction of subunits. The
secondary bonds involved in subunits interaction are loose than those in secondary and
tertiary structures and proteins with quaternary structure can dissociate to separate subunits
with their conformation unchanged under certain conditions.
Proteins 147

a
electrovalent bond;
b
hydrogen bond;
c
hydrophobic bond;
d
Vander Wasls force;
e
disulfide bond.

Figure 5-1. Secondary bonds involved in tertiary structure stabilization [1].

Figure 5-3 shows the combination pattern of the subunits of hemoglobin. The structures
of subunits in a protein may be the same or different. For example, the coat protein of tobacco
streak virus consists of 2200 identical subunits and normal hemoglobin A is a tetramer
containing two α subunits and two β subunits. The smallest subset of different subunits of a
protein is defined as the protomer. For example, the aspartate carbamyl transferase contains
six protomers and each protomer consists of one catalytic subunit and one regulatory subunit.
Some proteins can further aggregate into polymers and each repeating unit is called a
monomer. According to the number of monomers, protein polymers can be divided into
dimer, trimer, oligomer and multimer. For example, insulin might occur as dimer and
hexamer in human body.

Figure 5-2. Tertiary structures of myoglobin and triosephosphate isomerase [2].


148 Hong Lin, Lisha Wu and Shuhui Wang

Figure 5-3. Binding pattern of hemoglobin subunits [3].

2.3. Structure-Function Relationships

2.3.1. Relationship between Primary Structure, Conformation and Function


Primary structure determines the spatial arrangement of peptide chains and the
conformation is mainly maintained by various secondary bonds of side chains in amino acid
residues. Once a polypeptide chain is synthesized in vivo, the protein folds automatically to
the correct conformation.
Proteins with similar primary structure have similar conformations and functions. For
instance, proteins of same functions isolated from different organisms only differ slightly in
primary structure and the difference is much smaller between organisms with closer
evolutionary relationships.

2.3.2. Relationship between Spatial Conformation and Function


The diverse functions of proteins are closely related to their spatial conformations. Once
the conformations changed, the functional activities vary accordingly. The relationship
between conformation and function of proteins can be evidenced by protein denaturation and
renaturation, which will be detailed later.

3. CLASSIFICATION OF PROTEINS
3.1. Classification by Amino Acid Composition and Quantity

According to the composition of amino acids and their contents in proteins, proteins are
divided into complete, partially complete, and incomplete proteins.

3.1.1. Complete Protein


A complete protein (or whole protein) contains an adequate proportion of all nine of the
essential amino acids necessary for the dietary needs of humans or other animals. Many
animal products, such as milk, eggs, fish, and meat, contain complete proteins.
Proteins 149

3.1.2. Partially Incomplete Protein


A partially incomplete protein contains all the amino acid species, but their proportions
are insufficient for human demand. Partially incomplete proteins can maintain life, but cannot
enhance growth and development. For example, gliadin in wheat contains a low proportion of
Lys and is thus designated as a partially incomplete protein. Any essential amino acid that is
present in insufficient quantity for body protein synthesis is defined as a limiting amino acid.
Corn proteins contain low levels of Lys and thus Lys is the limiting amino acid.

3.1.3. Incomplete Protein


Incomplete proteins lack one or more essential amino acids in correct proportions as
necessary for good nutrition and health. Organism cannot grow, develop, or maintain their
lives only on incomplete proteins. Collagen in pork skin is an example of incomplete protein.

3.2. Classification by Solubility

Based on solubility, proteins are divided into seven categories.

3.2.1. Albumin
Albumin refers to any protein that is soluble in water and moderately soluble in
concentrated salt concentrations. Albumin can be precipitated by saturated ammonium
sulphate. Albumin occurs widely in nature. Leucosin in wheat seeds, blood plasma proteinand
ovalbumin in eggs are examples of albumin.

3.2.2. Globulin
Globulins are insoluble in water but soluble in diluted salt, acid, or alkaline solutions and
can be precipitated by half-saturated ammonium sulphate. Globulins have important
biological functions. Legumin in soybean seeds, seroglobulin in blood, myoglobulin in
muscle, and immunoglobulins are of this category.

3.2.3. Histone
Histones are highly alkaline proteins and are soluble in water or diluted acid solutions.
Histones contain rich quantity of Arg and Lys and are the structural components of
chromosomes.

3.2.4. Protamine
Protamines are small, arginine-rich proteins and are soluble in water and diluted acid
solutions. Protamines contain abundant alkaline amino acids and lack Trp and Tyr. The
proteins are found in mature sperm cells and replace histones late in chromosome the haploid
phase of spermatogenesis.

3.2.5. Prolamin
Prolamins are a group of plant storage proteins with high proline contents and found in
the seeds of cereal grains. They are characterized by a high glutamine and proline content and
are generally soluble only in 70%~80% ethanol solutions.
150 Hong Lin, Lisha Wu and Shuhui Wang

3.2.6. Gluten
Glutens are contained in plant seeds, such as barley and rye. Proteins of this category are
insoluble in water or diluted salt solutions but soluble in diluted acid and alkaline solutions.

3.2.7. Albuminoid
Proteins of this category are not soluble in water or salt, diluted acid and diluted alkaline
solutions and occur mainly in skins, hairs, and nails. The roles of such proteins include
protection and support, forming connective tissue, tendons, bone matrices, and muscle fiber.
Keratin, collagen, elastin, and fibroin are all albuminoids.

3.3. Classification by Chemical Composition

3.3.1. Nucleoprotein
A nucleoprotein is any protein which is structurally associated with nucleic acid.
Deoxyribonucleoprotein, ribosome, and tobacco mosaic virus are examples of nucleoprotein.

3.3.2. Lipoprotein
A lipoprotein is a complex molecule consisting of both protein and lipid. The lipid might
be phospholipids, sterols, or neutral fats. Many transporters in blood and lipovitellin are
lipoproteins.

3.3.3. Glycoprotein And Mucoprotein


In glycoproteins, proteins are covalently attached to oligosaccharides and the glycans
can be galactose, mannose, hexosamine, hexuronic acid, or many other sugars. Mucoprotein
is a typical example of glycoprotein and it is primarily composed of mucopolysaccharides.

3.3.4. Phosphoprotein
In phosphoproteins, phosphoric acid is connected to the Ser or Thr resides of proteins
through ester bonds. Casein and pepsin belong to this category.

3.3.5. Metalloprotein
Metalloproteins refer to any proteins that contain metal ion cofactors. Many important
enzymes, transport proteins and storage proteins are metalloproteins. For example,
hemoglobin contains iron, chlorophyl contains magnesium, and hemocyanin contains copper.

3.3.6. Flavoprotein
Flavoproteins are proteins that contain a nucleic acid derivate of riboflavin. Succinic
acid dehydrogenase and d-amino acid oxidase are examples of flavoprotein.
Proteins 151

4. PROTEIN DENATURATION
4.1. Definition of Denaturation

When a protein is exposed to such physical stresses as heating and ultraviolet irradiation
or such chemical stresses as strong acid and base, its properties are changed, such as decrease
of solubility and loss of activity, but the primary structure remains unaltered. This process is
termed denaturation. Denaturation is an extremely important property of proteins and
markedly affects food processing.

4.2. Mechanism of Denaturation

The denaturation theory was first proposed by a Chinese biochemist in 1931. It was
believed that denaturation is the transfer of natural proteins molecules from ordered and
compact states to disordered and loose states because of environmental stresses. The compact
structure of natural proteins is maintained by various secondary bonds. These secondary
bonds are easily interrupted by physical and chemical factors, which induce the destruction or
alternation of protein conformation. Hence, protein denaturation is actually caused by the
changes of secondary, tertiary, and quaternary structures of proteins. The destruction of
spatial conformation of proteins leads to reduced solubility, aggregation, irreversible gelation,
increased sensitivity to protease hydrolysis, and loss of physiological activities.
A protein denatured in mild conditions is present in a relaxed conformation. After the
denaturation factor is removed, the protein can restore to its natural conformation. Hence, this
denaturation process is reversible. For example, when ribonuclease is treated with 8 mol/L
urea and mercaptoethanol, the disulfide bonds in the molecule is reduced and the enzyme
loses its activity due to changed conformation. However, after the regents are removed by
dialysis, the denatured protein is automatically oxidized and returns to its natural
conformation (Figure 5-4).

Figure 5-4. Denaturation and renaturation of ribonuclease [4].


152 Hong Lin, Lisha Wu and Shuhui Wang

Trypsin and hemoglobin also undergo reversible denaturation. After trypsin is denatured
by heating at 70~100°C for a short time in an acidic solution, the enzyme loses its catalytic
activity. When the solution is cooled down in a proper rate, trypsin can recover all its
properties.
Haemoglobin can be denatured by low-concentration sodium salicylate. After sodium
salicylate is removed, the protein can restore to its natural conformation. However, the
denaturation of most proteins is irreversible. For example, egg protein is gelated when
cooking and this process is irreversible; soybean protein is irreversibly denatured in tofu
production.
The reversibility of protein denaturation is related to the denaturation agent species,
protein property and the degree of protein conformation destruction. Generally, in reversible
denaturation, the tertiary and quaternary structures of proteins are destroyed, but secondary
structures remain intact. In contrast, in the irreversible denaturation, the secondary, tertiary
and quaternary structures are all interrupted.

4.3. Properties of Denatured Protein

The physical, chemical and biological properties of proteins undergo obvious changes
after denaturation.

(1) Hydrophobic groups originally located inside molecules get exposed on molecule
surface and the hydration layer is destroyed. Hence, the solubility of denatured proteins
decreases significantly.
(2) Denatured proteins cannot crystallize as original ones.
(3) The spatial structure of denatured proteins is in random shape. The friction between
molecules increases and the fluidity decreases. Hence, denatured proteins have reduced
diffusion coefficient and their solutions have lower fluidity.
(4) The optical activity of denatured protein changes and the isoelectric point increases
slightly.
(5) Denatured proteins are more susceptible to enzymatic hydrolysis. For example,
natural haemoglobin cannot be hydrolyzed by trypsin, but denatured haemoglobin can be
hydrolyzed easily. This is why cooked foods are digested and absorbed more easily.
(6) Denatured proteins have more exposed side chains, such as –SH and –OH. Hence,
the degree of protein denaturation can be estimated by the reaction between these groups and
specific reagents.
(7) The bioactivities of denatured proteins are completely or partially destroyed. For
example, denatured enzymes lose their catalytic activities; denatured hemoglobin can no
longer transport oxygen; and denatured antigens lose their immunological competences.

4.4. Denaturation Agents and Their Mechanisms

4.4.1. Heat
Heating is the most common physical factor of protein denaturation. The temperature
required for detectable denaturation varies with the nature of protein and is also affected by
Proteins 153

protein purity and pH. The denaturation of most proteins can be detected in the temperature
range 45~50°C and the process is accelerated when the temperature is elevated to 55°C.
Denaturation occurring at lower temperatures involves only the disruption of non-covalent
bonds and proteins become unfolded. For example, natural serum albumin is of the oval shape
with the axial ratio of 3:1, while the axial ratio of denatured serum albumin increases to 5:5.
Such denaturation induced by short-time exposure to low temperatures is reversible. When
the temperature increases to 70~80°C, disulfide bonds are broken. Hence, protein
denaturation occurring in long-time high-temperature treatment is irreversible.
Heat treatment leads to protein denaturation by disrupting hydrogen bonding and many
other secondary bonds that maintain protein conformation. It is estimated that the rate of
denaturation increases by up to 600 folds when the temperature increases by 10°C in the
denaturation temperature range.
The sensitivity of a protein to thermal denaturation depends on multiple factors, such as
protein property, protein concentration, water activity, pH, ionic strength and ion species.
Proteins, enzymes, and microbes are more resistant to thermal denaturation in water-
containing conditions than in dry conditions. Besides, high concentration of denatured protein
hinders the renaturation process.
The temperature used in the thermal processing of foods is generally around 100°C and
the central temperature of materials often exceeds 70°C. Hence, most proteins are denatured
and pathogens are inactivated during thermal processing of foods.

4.4.2. Radiation
The impact of radiation on proteins depends on the energy and wavelength of the rays
used. Ultraviolet radiation can be absorbed by aromatic amino acid residues (tryptophan,
tyrosine and phenylalanine) and can change the conformation of proteins. Radiation by high-
energy rays disrupts disulfide bonds. γ radiation and other ionizing radiation can also make
conformation changes, amino acid oxidization, covalent bond disruption, ionization, proteins
free radical formation as well as polymerization.

4.4.3. Denaturation at Interfaces


The denaturation of proteins attached to water-air, aqueous-nonaqueous solution, and
water-solid interfaces is generally irreversible. Proteins are surfactants and tend to migrate to
the interface and be adsorbed. The adsorption rate of proteins is controlled by diffusion rate of
natural proteins to the interface and the absorption stops when the interface is saturated by
denatured proteins (about 2mg/m2). Figure 5-5 indicates that the migration and absorption of
globular proteins in the aqueous-nonaqueous interface.
During the migration to interfaces, proteins interact with high-energy water molecules
(compared with the water molecules far away from the interface) at the interface. The
hydrogen bonding between protein molecules is then disrupted and the protein conformation
becomes slightly unfolded (Figure 5-5b). Because more hydrophobic groups are in contact
with water, the partially unfolded protein is hydrated and activated and transforms to an
unstable state. The protein further extends and unfolds at the interface and the hydrophobic
and hydrophilic residues are positioned in the water and non-aqueous phases respectively,
leading to protein denaturation at the interface. Proteins that are mainly stabilized by disulfide
bonds are not readily absorbed at the interface. Hence, these proteins are less susceptible to
denaturation at interface.
154 Hong Lin, Lisha Wu and Shuhui Wang

a)

b)

c)

Figure 5-5. Schematic diagram of protein interface denaturation [5].

The interfacial properties of proteins are important for their applications in various food
systems. For example, proteins absorbed at interfaces are beneficial for the formation and
stabilization of emulsions and foams. To keep the natural conformation and functional
properties of proteins, dispersion systems, such as foams and emulsions, should be avoided
during food processing and separation.

4.4.4. pH
It is irreversible that protein denaturation occurs in mild and extreme pH conditions.
Extreme pH changes the ionization of some groups in polypeptide chains and disrupts the
hydrogen bonding required for maintaining conformation and the electrostatic interaction
between differently charged groups. If the pH is lowered far below the isoelectric point (pI) or
elevated above the pI, the protein will contain only positive or negative charges. The charges
will repel each other and prevent the protein from aggregation. In areas with large charge
density, the intramolecular repulsion may be great enough to cause unfolding of the protein.
In some cases the unfolding may be extensive enough to expose hydrophobic groups and
Proteins 155

cause irreversible aggregation. Yogurt is a typical example of acid-induced protein


denaturation.

4.4.5. Heavy Metals


Heavy metal salts containing Hg2+, Pb2+, Ag+, T+, Cd2+ and other metals with high
atomic weights can disrupt the salt bridges and disulfide bonds in proteins. The reaction of a
heavy metal salt with a protein usually leads to an insoluble metal-protein salt.
This reaction is used for its disinfectant properties in external applications. For example
AgNO3 is used to prevent gonorrhea infections in the eyes of new born infants. Silver nitrate
is also used in the treatment of nose and throat infections, as well as to cauterize wounds.

Figure 5-6. Denaturation of prion protein by alcohol.

Figure 5-7. Denaturation of prion protein by reducing agents.


156 Hong Lin, Lisha Wu and Shuhui Wang

This same reaction is used in reverse in cases of acute heavy metal poisoning. In such a
situation, a person may have swallowed a significant quantity of a heavy metal salt. As an
antidote, a protein such as milk or egg whites may be administered to precipitate the
poisonous salt. Then an emetic is given to induce vomiting so that the precipitated metal
protein is discharged from the body.

4.4.6. Chemical Reagents


Most organic solvents can cause protein denaturation. Organic solvents disrupt the
hydrogen bonding between water and proteins, which leads to change the dielectric constant
of medium. Non-polar organic solvents can enter the hydrophobic area of proteins and
denature proteins by destroying the hydrophobic interaction. A 70% alcohol solution is often
used as a disinfectant on the skin. Alcohol denatures proteins by disrupting the side chain
intramolecular hydrogen bonding. New hydrogen bonds are formed instead between alcohol
and protein side chains. In the prion protein, Tyr 128 is hydrogen bonded to Asp 178, which
cause one part of the chain to be bonding with a part some distance away. After denaturation,
the graphic show substantial structural changes, as shown in Figure 5-6.
Urea denatures protein also by disrupting the hydrogen bonding, because it forms new
hydrogen bonds with proteins. Guanidine hydrochloride denatures proteins in the similar
mechanism as urea.
Different protein chains or loops within a single chain are often held together by the
strong covalent disulfide bonds. Reducing agents, such as dithiothreitol, can split disulfide
bonds apart and cause protein denaturation. Figure 5-7 shows the denaturation of proteins by
reducing agents.

4.5. Effects of Combined High-Pressure and Thermal Treatment on Beef


Protein Denaturation

Thermal treatment in combination with high pressure improves the tenderness of beef
and enhances the sterilization effect. Besides, the combined treatment changes the
composition of beef muscles and affects its functional properties, such as color, texture, fat
oxidation and flavor, which are related to protein denaturation. Differential scanning
calorimetry(DSC) is an effective tool for studying the structural and thermodynamic
properties of natural polymers. DSC has been used in the investigation of denaturation and
resultant texture changes of muscle proteins. Figure 5-7(a) Figure 3 shows the thermograms
of intact beef muscle after treatment at pressures from 0.1 to 800MPa at 20°C for 20min. The
three major endothermic transitions seen have been attributed to myosin, collagen and actin
and the peak maxima in this study are seen at 54.6°C (myosin), 67.1°C (collagen) and 77.3°C
(actin). Treatment at 200MPa decreased the myosin and actin peaks and at 400MPa and
higher, the actin peak is not seen.
Figure 5-7(b) shows the thermograms when the samples are treated at 0.1-800MPa at
40°C for 20min. At ambient pressure about 43% of the myosin, is denatured but there is no
obvious effect on collagen and actin. At 200MPa the myosin and collagen are merged into
one broad peak with an onset temperature of 50.1°C. At 400MPa, the actin peak is not seen.
When the pressure is increased to 600 and 800MPa, the new myosin peak and that of collagen
are still discernible.
Proteins 157

a) Peak 1 Peak 3
Peak 2
0.2 mw

0.1 MP2

Peak N
200 MP2

Heat Flow (mw)


400 MP2

600 MP2

800 MP2

Rescan

50 55 60 65 70 75 80 85
Temperature (˚C)

b)

40 45

Figure 5-7. Thermograms of beef muscle, heated at 10 ˚C min−1 after treatment at different pressures in
air for 20min at 20°C and 40°C [7].

Although the combination of high pressure and thermal treatment has complex effect on
protein properties, it has wide applications in food processing:

Improving texture and tenderizing by causing protein unwinding and aggregation;


Extending shelf life by inactivating enzymes, microbes, and toxins activity;
Increasing digestibility;
Increasing the ability of maintaining flavors, pigments and vitamins by increasing the
surface hydrophobicity due to unwinding;
Improving product flavor and overall acceptability.

4.6. Effects of Freezing on Aquatic Protein Denaturation

Frozen fish alter in characters during storage at sub-zero temperatures, becoming


progressively tougher to eat, and extruding much fluid or drip on thawing. The deteriorative
changes in texture as a consequence of long-term storage are considered to be due to protein
denaturation during frozen storage and the change proceeds more slowly in low temperature.
Most of these observed changes have been attributed to the changes of proteins in low
temperatures.
158 Hong Lin, Lisha Wu and Shuhui Wang

5. FUNCTIONAL PROPERTIES OF PROTEINS


The functional properties important for the food processor are attributes, which at the
proper concentration of the respective components or additives and at appropriate conditions,
provide for the desirable characteristics of the product. These properties of proteins are
displayed in interactions with the surrounding solvent, ions, other proteins, saccharides,
lipids, and numerous other components, as well as in surface phenomena (chemical and
functional properties of food components). Table 5-6 lists the functional properties of proteins
involved in different food systems.

Table 5-6. Functions of proteins in different food systems [8]

Functionality Food Systems Proteins involved


Solubility Beverage Lactalbumin
Viscosity Soup, sauce, salad dressing, dessert Gelatin
Water holding Sausage, cake, bread Muscle protein, egg protein
Gelation Meat, gell, bakery product, cheese Musscle protein, egg protein, milk
protein
Conglutination Meat, sausage, noodle, bakery product Muscle protein, egg protein,
lactalbumin
Elasticity Meat, bread Musscle protein, cereal protein,
milk protein
Emulsification Sausage, bologna sausage, soup, cake, Muscle protein, egg protein,
dessert lactalbumin
Foamability Ice cream, cake, dessert Egg protein, lactalbumin
Fat and flavor binding Low-fat bakery product, doughnut Milk protein, egg protein, cereal
protein

The sensory properties of foods result from the complex interaction between various
food ingredients. For example, the flavor, texture, color and shape of a cake is contributed by
the combination of the thermal gelation capacity, foaming capacity, water holding capacity,
emulsification, elasticity and browning reaction of the materials used. Hence, proteins as food
materials must have multiple functional properties (Table 5-7). Animal proteins, such as those
derived from milk, egg and meat, are mixture of multiple protein components and thus have
diverse physical and chemical properties. For example, egg white has good water holding,
gelling, conglutination, emulsification, foaming, and thermal coagulation capabilities and
these properties are contributed by ovalbumin, conalbumin, ovomucin, lysozyme and other
albumins and their interactions. Plant proteins, such as those from soybean and oilseeds, are
also mixtures of multiple protein components.
However, these proteins have only limited functional properties and are only used in
common foods in less quantity.

5.1. Interfacial Properties of Proteins

Bakery products, desserts, beer, milk, ice cream, butter, chopped meat, and other foods
containing emulsions or foams are dispersion systems and amphiphilic components must be
present to stabilize the systems. Proteins are amphiphilic molecules and can spontaneously
Proteins 159

migrate to the air-water or oil-water interface and form highly viscoelastic film at the
interface. The interface system of proteins is more stable than that formed by surfactants.

Table 5-7. Functional properties of proteins required for various food [9]

Food Functionality required


Beverage, soup, sauce Solubility in solutions of different pH, thermal stability, viscosity,
emulsifying and water holding capacity
Formed bakery products Casting and viscoelastic film forming, coherence, thermal denaturation,
gelation, swelling, emulsification, foaming, browning reaction
Dairy products Emulsification, fat retention, viscosity, foaming, gelation, coagulation
The substitute for egg Foaming, gelation
Meat products Emulsifying, gelation, coherence, absorption and retention of water and
fat
Meat extender Absorption and maintain of water and fat, insolubility, hardness,
chewiness, coherence, thermal denaturation
Food film Coherence, adhesion
Dessert Dispersity, emulsification

In addition to the intrinsic properties, including amino acid composition, structure,


conformation, distribution and ratio of hydrophobic and hydrophilic groups, number of
disulfide bonds, and shape, size, and flexibility, the interfacial properties of proteins are also
affected by environmental conditions, such as pH, temperature, ionic strength, salt species,
interface composition, protein concentration, presence of saccharides and low-molecular
weight surfactants, and energy input as well as container and operation sequence.
A protein as an ideal surfactant should has the following three attributes: moving rapidly
to the interface; extending and directing rapidly once reaching the interface; and forming
films of strong cohesion and viscoelasticity with neighboring molecules. Hence, the
distribution of hydrophobic and hydropholic groups in proteins and protein shape markedly
affect the interfacial properties. Supposing the surface of a protein is highly hydrophilic and
no discernable hydrophobic domain is found on the surface, the protein has a positive
adsorption free energy. That is, the free energy of the protein in the aqueous phase is lower
than in the interface or non-aqueous phase and the adsorption dose not occur. In contrast, if
the protein contains multiple hydrophobic domains on the surface, the protein can be
absorbed on the interface spontaneously, as shown in Figure 5-8. In another word, absorption
to the interface occurs only when the number of hydrophobic areas on the surface is sufficient
for providing the energy for hydrophobic domain-interface interaction.
The hydrophobic and hydrophilic groups of a protein must be positioned correctly at the
interface for the formation and stabilization of foams and emulsions. Generally, most proteins
are distributed in the bulk phase and only a small proportion is absorbed at the interface. High
flexibility facilitates the adsorption and positioning of proteins at the interface. For example,
casein has very good flexibility. Once casein is absorbed at the interface, conformation
change occurs rapidly and more peptide segments are absorbed at the interface. Rigid
globular proteins, such as lysosome and soybean protein, do not undergo marked
conformation transition at the interface.
Flexible polypeptide chains might exhibit three typical conformations at the interface:
train, tail, and loop, as shown in Figure 5-9. A flexible protein can be present as one or more
of these conformations at the interface. Generally, peptide segments in directly contact with
160 Hong Lin, Lisha Wu and Shuhui Wang

the interface occur in the train conformation, the segments suspended in the aqueous phase
are arranged as loops, while the peptide chains with the N- and C- terminal in the aqueous
phase are present mainly in the tail conformation. Of the three conformations, the train
conformation occurs in higher probability than the other two conformations and exhibits
lower interfacial intension.

)NTERFACE

-ODERATEPROBABILITY 3TRONGPROBABILITY
.OADSORPTION OFADSORPTION OFADSORPTION

(YDROPHILIC .ONPOLARSURFACE
SURFACE
!QUEOUSPHASE
(YDROPHOBICCORE

Figure 5-8. Probability of protein adsorption to the interface as a function of hydrophobic domain
number [11].

1IBTF

USBJO
*OUFSGBDF

UBJM
MPPQ

1IBTF

Figure 5-9. Configuration of flexibility polypeptide on interphase [12].

The stability of a protein at the interface determines the mechanical strength of the film
formed at the interface and the film strength is also affected by the electrostatic attraction, and
hydrogen bonding, and hydrophobic interaction between molecules. Disulfide bonds increase
the film viscoelasticity. When the concentration of protein in the interfacial film reaches
about 20%~25% (W/V), gelation occurs. To maintain the stability viscoelasticity of the gelled
film, the secondary interactions involved must be in balance. If the hydrophobic interaction is
too strong, the protein flocculates, aggregates, and even precipitates. If the electrostatic
repulsion far exceeds electrostatic attraction, a thick cohesive membrane is not formed
readily.

5.1.1. Emulsifying Properties


Proteins play important role in the stabilization of many food emulsion systems, such as
milk, butterfat, ice cream, and soy milk. Proteins are less capable of stabilizing W/O
emulsions than O/W emulsions, because most proteins are highly hydrophilic and most
absorbed proteins are located in the aqueous phase.
Proteins 161

Table 5-8. Emulsifying capability index (EAI) of some proteins [12]

Protein EAI (m2/g) Protein EAI (m2/g)


pH6.5 pH8.0 pH6.5 pH8.0
BSA 197 hemoglobin 75
Casein ache sodium 149 166 Yeast protein 8 59
β-lactoglobulin 153 lysozyme 50
Powder form lactalbumin 119 142 Ovalbumin 49
Note: The data were obtained when proteins are dispersed in 0.5% PBS buffer of ionic strength of 0.1.

The emulsifying capability of a protein can be evaluated by droplet size and distribution,
emulsifying activity, emulsion capacity, and emulsion stability.
Emulsifying activity refers to the total interfacial area of an emulsion. The emusifying
activity of a protein is often presented as the emulsifying activity index (EAI), namely the
interfacial area created per unit mass of protein. The EAI of a protein can be determined
easily in the turbidimetric method. The turbidity of an emulsion can be calculated by the
following equation:

A is the absorbance of the solution and L is the path length.


According to the Mie theory of light scattering, the interfacial area of an emulsion is two
and C is the weight of protein per unit
volume of the aqueous phase, then the EAI of the protein can be calculated by:

Emulsion capacity (EC) is the volume (ml) of oil that can be emulsified by per gram of
protein before phase inversion (a change from oil-in-water emulsion to water-in-oil) occurs.
In the determination of TC, oil or melted fat are added at a constant rate and temperature to an
aqueous protein solution that is continuously agitated. Phase inversion is detected by an
abrupt change in viscosity or color (usually a dye is added to the oil).
EMULSIFYINGACTIVITYINDEX%!)-G










         
$(
Figure 5-10. Effect of molecular weight on EAI (DH is the degree of hydrolysis) [13].
162 Hong Lin, Lisha Wu and Shuhui Wang

Note: ○ - 0.1mol/L NaCl; ● – 0.2 mol/L NaCl; ▲ – 0.5mol/L NaCl; □ – 1.0mol /L NaCl.

Figure5-11. Effect of pH and NaCl concentration on the emulsion volume of peanut protein isolate [14].

The emulsion stability (ES) is measured as the final volume of the emulsion after
centrifuging the initial volume or standing for several hours at specified conditions. It may
also be determined as the quantity of oil or cream separated from the emulsion, or the time
required for the emulsion to release a specified quantity of oil.

The properties of protein-stabilized emulsions are influenced by many factors. Intrinsic


factors are pH, ionic strength, temperature, presence of low-molecular-weight surfactants,
sugars, oil-phase volume, type of protein, and the melting point of the oil used. Extrinisic
factors include type of equipment, rate of energy input, and rate of shear. For example, the
emulsion capacity or emulsion stability of proteins is generally positively correlative with
protein concentration in the range 25~80%. Molecular weight also affects the EAI, as shown
in Figure5-10. The addition of sodium chloride increases the emulsion volume of meat
stuffing (Figure 5-11). Proteins exhibit different emulsifying activities at their isoelectric
points (pI). Proteins have minimum solubility at pI and hence have decreased emulsifying
activities. On the other hand, proteins are present in a highly viscoelastic structure at their pI.
This structure hinders the unfolding or adsorption at the interface, which is not beneficial for
emulsion formation, but can stabilize the film that is formed by already absorbed proteins.
Hence, proteins might have increased emulsifying activity under certain conditions. Collagen,
serum protein and ovalbumin have the maximum emulsifying activities at their pI, but
soybean protein, peanut protein, casein, lactalbumin, bovine serum albumin and troponin
have the maximum emusifying activites at other pH than their pI.

5.1.2. Foaming Properties


Food foams are dispersions of gas bubbles in a continuous liquid or semisolid phase.
Foaming is responsible for the desirable rheological properties of many foods, such as the
texture of bread, cakes, whipped cream, ice cream, and beer froth. In most of these products,
proteins are the main surface-active agents that help in the formation and stabilization of the
dispersed gas phase. Thus foam stability may be an important food quality criterion.
However, foams are often a nuisance for the food processor, such as in the production of
potato starch or sugar.
Proteins 163

In most cases, the disperse phase is air or carbon dioxide and the continuous phase is
composed of two films of proteins adsorbed on the interface between a pair of air bubbles,
with a thin layer of liquid between them. The diameters of bubbles might range from 1μm to
several centimeters, depending on the surface tension and viscosity of the aqueous phase and
energy input. Evenly distributed small bubbles can impart foods with desired consistency,
exquisite texture, and softness and improve the dispersivity and flavor. The formation of
bubbles is shown in Figure 5-12.

# &
%

$
"

LIQUID GASLATEX BUBBLE

Figure 5-12. Formation of bubbles [15].

Bubbles can also be formed by whipping or violently shaking a protein-containing


solution in the presence of gas. Whipping is the most common measure used to produce
bubbles. Compared with the first method mentioned above, whipping can produce stronger
mechanical stress and shearing force and hence gases are dispersed more uniformly.
However, too strong mechanical stress might affect the aggregation and formation of bubbles
by hindering the absorption of protein at the interface. Besides, the amount of proteins
required for stabilizing bubbles is also increased by 1%~40% (w/v). The volumes of materials
can increase by 300%~2000% after whipping.
The third method of producing bubbles is to suddenly release the pressure of a pressured
solution. This method has been used in the production of whipped butter.
The foaming power of proteins and the stability of bubbles foams are affected by many
factors, such as pH, salts, saccharides, lipids, and protein concentration. Table5-9 lists the
foaming power of some proteins.

Table 5-9. Foaming capability of protein [16]

Protein type Foaming power at 0.5% Protein type Foaming power at


protein concentration 0.5% protein
(W/V) concentration (W/V)
Bovine serum 280 β-Lactoglobulin 480
albumin
Whey protein 600 Fibrinogen 360
isolate
Ovalbumin 40 Soybean protein (enzyme 500
hydrolyzed)
Egg albumen 240 Gelatin (acid-processed 760
pig skin produce)

The pH is an important factor that affects the foaming ability and foam stability of
proteins. Generally, high solubility is a prerequisite for good foaming ability and foam
stability. However, insoluble protein granules, such as fibrillin in its isoelectric point, may
164 Hong Lin, Lisha Wu and Shuhui Wang

impart good foam stability. Some proteins give rise to small to foam volume in their
isoelectric points, but the produced foams have excellent stability, such as globulins in pH
5~5, glutelin in pH 6.5~7.5, and lactalbumin in pH 4~5. Some protein-stabilized foams are
more stable in extreme pH values, possible due to increased viscosity of involved proteins.
The foams in most foods are formed in other pH than the isoelectric points of contained
proteins.
Sugars suppress the expansion of protein-stabilized foams, but increase the foam
stability. This is because sugars can increase the viscosity of the bulk phase and reduce the
drainage of the lamella fluid. Sugars can stabilize the conformation of proteins and hinder
their adsorption and unfolding on the interface. Hence, proteins cannot produce large
interfacial area and foam volume in the presence of sugars. This is why sugars are often
added after the foam expansion has already been completed in the manufacture of meringue
and other sugar-containing foods.
Lipids in low concentrations (about 0.1%) seriously impair the foam ability of proteins.
This is because lipids are more surface-active than proteins. Lipids readily adsorb at the air-
water interface and inhibit adsorption of proteins during foam formation. Besides, lipid films
lack the cohesive and viscoelastic properties necessary to withstand the internal pressure of
the foam bubbles, the bubbles rapidly expand, then collapse during whipping. Thus, lipid-free
whey protein concentrates and isolates, soy proteins, and egg proteins without egg yolk
display better foaming properties than do lipid-contaminated preparations.
Partially denatured proteins have improved foaming abilities than their nature forms.
Moderate heat treatment can enhance the foaming ability of soybean protein (70~80°C) and
lactalbumin (40~60°C). Although moderate heat treatment increases the expansion capacity,
it decreases the foam stabilization. Excessive heat treatment might impair the foaming ability
of proteins.
To obtain enough quantity of foam, the whipping operation must last long enough in
suitable strength for sufficient unfolding and adsorption. However, excessive whipping can
reduce the expansion volume and foam stability. For example, ovalbumin is very sensitive to
excessive whipping. Whipping of ovalbumin for 6~8min can lead to coagulation-flocculation
in the air-water interface. These insoluble proteins could not be completely absorbed on the
interface and the viscosity of the formed liquid sheet cannot meet the requirement of high
stability of foams.
Most proteins contain more than one subunit. Hence, the foaming ability of a protein is
also affected by the interaction between the subunits absorbed on the interface. For example,
the excellent foaming ability of egg whit has been attributed to its protein composition.
Acidic proteins can get improved foaming ability when complexing with an alkaline protein.
No close correlation between emulsifying ability and foaming ability has been found for
protein.
Salts affect the foaming of proteins by influencing the solubility, viscosity, unfolding
and aggregation of proteins. The foaming ability and foam stability of most globular proteins,
such as bovine serum albumin, egg albumin, gluten and soy proteins increase with increasing
concentration of NaCl. This behavior is usually attributed to neutralization of charges by salt
ions. However, some proteins, such as whey proteins, exhibit the opposite effect: Both foam
ability and foam stability decrease with increasing concentration of NaCl (Table 5-10).
Bivalent cation ions, such as Ca2+ and Mg2+, in concentration 0.02~0.04mol/L could bridge
Proteins 165

with the carboxyl group of proteins to form protein films with good viscoelasticity. Hence,
the presence of these ions can increase the foam stability.

5.2. Viscoelasticity

Viscoelasticity is the property of materials that exhibit both viscous and elastic
characteristics when undergoing deformation.

Table 5-10. Effect of NaCl on the foaming ability and foam stability of whey protein
isolate [17]

Concentration of Total interfacial area (cm2/mL of Times of 50% collapse of initial area
NaCl(mol/L) foam) (second)
0.00 333 510
0.02 317 324
0.04 308 288
0.06 307 180
0.08 305 165
0.10 287 120
0.15 281 120

Wheat protein is very unique among food proteins because it can produce dough with
both viscous and elastic properties. When a mixture of wheat flour and water (about 3:1 ratio)
is kneaded, a dough with viscoelastic properties forms that is suitable for making bread and
other bakery products.
These unusual dough characteristics are mainly attributable to the proteins in wheat flour.
Wheat flour contains 20% soluble and 80% insoluble protein fractions. The soluble proteins
are primarily albumin, globulin-type enzymes and minor glycoproteins. These proteins do not
contribute to the dough-forming properties of wheat flour. The major storage protein of wheat
is gluten. Gluten is a heterogeneous mixture of proteins, mainly gliadins (soluble in
70%~90% ethanol) and glutenins (insoluble in water or ethanol, but soluble in acidic and
alkaline solutions). Gluten can entrap gas during fermentation to produce the viscoelastic
property. Besides, the starch granules, pentosan, polar and non-polar lipids, and soluble
proteins in wheat flour facilitate the formation of the viscoelastic protein network and dough
texture. Gluten is rich in Gln (more than 33% of the amino acid residues) and hydroxyl-
containing amino acids. Hence, the amino acids can form hydrogen bonding readily and this
property contributes to the water adsorption and cohesion-adhesion property of gluten. About
30% of gluten's amino acid residues are hydrophobic, and the residues contribute greatly to its
ability to form protein aggregates by hydrophobic interactions and to bind lipids and other
nonpolar substances.
Cysteine and cystine residues account for 2~3% of gluten's total amino acid residues.
During formation of the dough, these residues undergo sulfhydryl-disulfide interchange
reactions resulting in extensive polymerization of gluten proteins.
Several physical and chemical transformations occur during mixing and kneading of a
mixture of wheat flour and water. Under the applied shear and tensile forces, gluten proteins
absorb water and partially unfold. The partial unfolding of protein molecules facilitates
hydrophobic interactions and sulfhydryl-disulfide interchange reactions, which result in
166 Hong Lin, Lisha Wu and Shuhui Wang

formation of thread-like polymers. These linear polymers in turn are believed to interact with
each other, presumably via hydrogen bonding, hydrophobic associations, and disulfide cross-
linking, to form a sheet-like film capable of entrapping gas.
Because of these transformations in gluten, the resistance of the dough increases with
time until a maximum is reached, and this is followed by a decrease in resistance, indicative
of a breakdown in the network structure. The breakdown involves alignment of polymers in
the direction of shear and some scission of disulfide cross-links, which reduces the polymer
size. The time it takes to reach maximum dough strength during kneading is used as a
measure of wheat quality for bread making—a longer time indicating better quality.
The development of the viscoelastic dough is thought to be related to the extent of
sulfhydryl-disulfide interchange reactions. This is supported by the fact that when reductants,
such as cysteine, or sulfhydryl blocking agents, such as N-ethylmaleimide are added to
dough, viscoelasticity decreases greatly. On the other hand, addition of oxidizing agents, such
as iodate and bromate, increase the elasticity of the dough. This implies that wheat varieties
that are rich in SH and S-S groups might possess superior bread-making qualities, but this
relationship is unreliable. Thus, the role of sulfhydryl-disulfide interchange reactions in the
development of viscoelastic dough is poorly understood.
The strength of dough is related to the contents of glutenin and completely insoluble
residue proteins. Experiments on wheat flour supplemented with glutenins and gliadins of
different proportions indicate that glutenins determine the elasticity, cohesiveness, mixture
tolerance, extensibility, the expansion of dough and the mobility of gliadins. Suitable
proportions of the two categories of proteins are important for bread making.
High glutenins contents can increase the cohesiveness of dough and suppress the
expansion of entrapped CO2, while high gliadins contents increase the elongation and make
the gluten film fragile and permeable, leading to poor CO2 retention and bread collapse.
The different effects of glutenins and gliadins on bread-making may be related to the
differences in their structure and composition. In gluten these exist as single polypeptides
with molecular weight ranging from 30,000 to 80,000. Although gliadins contain about 2~3%
half-cystine residues, they apparently do not undergo extensive polymerization via
sulfhydryl-disulfide interchange reactions. The disulfide bonds appear to remain as
intramolecular disulfides during dough making. Dough made from isolated gliadins and
starch is viscous but not viscoelastic.
Glutenins are heterogeneous polypeptides with molecular weight ranging from 12,000 to
130,000. These are further classified into high-molecular-weight (M.W. >90,000, HMW) and
low-molecular-weight (M.W. <90,000, LMW) glutenins. In gluten, glutenin polypeptides are
present as polymers joined by disulfide cross-links, with molecular weights ranging into the
millions. Because of their ability to polymerize extensively via sulfhydryl-disulfide
interchange reactions, glutenins contribute greatly to the elasticity of dough. Therefore, an
optimum ratio of gliadins and glutenins seems to be necessary to form viscoelastic dough.
Some studies have shown a significant positive correlation between HMW glutenin content
and bread quality in some wheat varieties, but not in others. Available information indicates
that a specific pattern of disulfide crosslink association among LMW and HMW glutenins in
gluten structure may be far more important to bread quality than the amount of this protein.
For example, association/polymerization among LMW glutenins which have the similar
structure form the HMW gliadin. This type of structure contributes to viscosity of the dough,
but not to elasticity. In contrast, if LMW glutenins link to HMW glutenins via disulfide cross-
Proteins 167

links (in gluten), then this is believed to contribute to dough elasticity. It is possible that in
good-quality wheat varieties, more of the LMW glutenins may polymerize with HMW,
whereas in poor-quality wheat varieties, most of the LMW glutenins may polymerize among
themselves. These differences in associated states of glutenins in gluten of various wheat
varieties may be related to differences in their conformational properties, such as surface
hydrophobicity, and reactivity of sulfhydryl and disulfide groups.
Baking does not lead to the re-denaturation of gluten, because glutenins and gliadins
occur naturally in partially unfolded form and are further unfolded during kneading. When
heated in 70~80°C, gluten loses water and the water is absorbed by gelatinized starch
granules. Hence, gluten can maintain the softness and water content (40%~50%) of bread in
baking.
Soluble protein fractions, including albumin- and globulin-type proteins, are denatured
and coagulated during baking. This partial gelling is beneficial for the cohesion of crumbs.
Hence, supplementation of external proteins is good for bakery foods quality, such as
nutrition fortification. However, not all proteins are beneficial for gluten network formation.
For example, supplementation of wheat flour with albumin- and globulin-type proteins, such
as whey proteins and soy proteins, adversely affects the viscoelastic properties and baking
quality of the dough. These proteins decrease bread volume by interfering with formation of
the gluten network. Addition of phospholipids or other surfactants to dough counters the
adverse effects of foreign proteins on loaf volume. In this case, the surfactant/protein film
compensates for the impaired gluten film. Although this approach results in acceptable loaf
volume, the textural and sensory qualities of the bread are less desirable than normal.

5.3. Gelation

Gelation is the process of aggregation and ordered network formation of denatured


proteins. The gelation of proteins is very important for some foods, including various dietary
products, jelly, gelatin gel, soybean protein gel, and textured vegetable proteins. Tofu, which
is a favorite food in China, is a typical food prepared by protein gelation.
Heating is necessary for the formation of most protein gels. The addition of salts,
especially those containing Ca2+, improves the gelation rate and gel strength. Proteins can
also interact with polysaccharides to form gels. For example, the positively-charged gelatin
can bind to negatively-charged sodium alginate through nonspecific ionic interaction to form
gel with a high melting point (80°C).
Many protein gels are highly hydrated and each gram of protein can hold up to more
than 10 grams of water. Many food components can be trapped in the network of protein gels.
The water contents of some protein gels might reach up to 98%. Although the trapped water
has properties similar to the water in dilute solutions, the water cannot be squeezed out easily.

5.4. Hydration

The conformation of each protein in a solution is affected by their interactions with


water. The interaction between proteins and water also occurs in solid foods. In addition, dry
concentrated proteins or protein isolates must be hydrated before use. Hence, the hydration
168 Hong Lin, Lisha Wu and Shuhui Wang

and rehydration properties of proteins markedly affect their applications in the food industry.
Figure 5-13 illustrates the sequence of interactions occurred between dry protein and water.

Water molecules are


Dry Multilayer Liquid water Protein starts
absorbed by binding
protein water is formed condenses swelling
to polar groups

Protein
Solution
disperses in
is formed
water

Figure 5-13. Sequence of the interactions between dry protein and water [10].

The protein-protein and protein-water interactions are affected by environmental


conditions, including protein concentration, pH, temperature, time, ionic strength and the
presence of other components. The pH alternation affects the ionization and net charge of
proteins and thus changes the attraction and repulsion between proteins and their water
binding capacity. The water binding capability of proteins decreases with the increase of
temperature. High temperature decreases the bonding force between hydrogen bonds and
leads to protein denaturation and aggregation. Protein denaturation or aggregation reduces the
surface area of the protein and consequently the hydration of polar amino acids. When
proteins with compact strength are heated, the proteins are unfolded and the peptide chains
and polar chains that are originally located inside the structure are exposed to water and the
hydration capability is increased as a result. The gelation of some proteins, such as
lactoalbumin, is irreversible. If the gels of these proteins are dried, the gel network is
transformed to the capillary of the protein powder. The dried proteins thus have significantly
increased hydration capability. However, the rate and degree of hydration are also affected by
the size and the holes on the surface and inner of proteins. Ions might compete for the side
chains of amino acids with water and subsequently influence the hydration capability,
swelling property, and solubility of proteins. Salts in low concentrations enhance the
hydration capability of proteins, but salts in high concentrations can lead to dehydration due
to the excessive water-salt interaction over the water-protein interaction.

5.5. Solubility

It is a long process for proteins to reach their equilibrium solubility and the protein
solubility varies with the pH, ionic strength, and temperature of the environment and protein
concentration. The solubility of most proteins decreases irreversibly when heated. However,
heat treatment is unavoidable during food processing and protein denaturation might occur
during extraction and purification even in mild conditions. Hence, the nitrogen solubility
index (NSI) of soybean protein powder, concentrated soybean protein, and soybean protein
isolate vary in the range 10%~90%.
Proteins 169

The solubility of proteins is essential for determining the optimum conditions of protein
extraction, purification and separation and is an important indicator of its application range.
The degree of insolubility can be used as a measure to evaluate the denaturation and
agglutination of proteins. It is reported that proteins with greater initial solubility can disperse
quickly and yield well dispersed hydrocolloid system with macrostructure and smooth
texture. In addition, high initial solubility facilitates the diffusion between of proteins to the
gas/water and oil/water interface for enhanced their surface activity.

5.4. Viscosity

The viscosity or consistency of a liquid reflects its resistance to flow under an applied
force or shear stress and can be represented in viscosity coefficient η, which is defined as the
ratio of shear stress to shear rate (or flow rate). The viscosity of protein fluids is largely
related to the apparent diameter of dispersed protein molecules or protein particles. The
apparent diameter of proteins is decided by the following parameters: (1) the inherent nature
of protein molecules, such as molar concentration, molecular size, molecular volume,
molecular structure and charge; (2) protein-solvent interaction, which affects the swelling and
solubility of proteins and the hydrokinetics of the fluid surrounding protein molecules; (3)
protein-protein interaction. The interaction determines the size of protein aggregates and the
interaction opportunity increases in high protein concentrations.
The viscosity coefficient of protein-containing solutions, such as homogenates,
emulsions, pastes and gels, might decrease with the increase of shearing stress. This behavior
is known as shear thinning. During shearing, protein molecules are redirected to the flow
direction and the friction drag is reduced as a result. Besides, the break of hydrogen bonds
and other weak bonds can cause the dissociation of protein aggregates and network. The
break of weak bonds is a slow process so that the shear stress and apparent viscosity
decreases with the proceeding of shearing before the protein fluid reaches steady flow. When
the shearing force is no longer applied, the original aggregates or network can or cannot be
reformed. If the aggregates or networks can be completely or partially reformed, the change
of viscosity coefficient is irreversible.
The viscosity or consistency of protein system is an important property for liquid and
semisolid-type foods such as beverages, broth, sauces and cream. The fluid property of
protein dispersion systems is of practical significance in determining the optimum operation
conditions. For example, during transfer, blending, heating, cooling, and spray drying, the
fluidity of protein dispersion systems significantly affects the transfer of both mass and heat.

6. PROPERTIES OF FOOD PROTEIN MATERIALS AND


NEW PROTEIN DEVELOPMENT
6.1. Muscle Protein

Muscle derived from mammals, birds and fish is the main source of meat. The skeletal
muscle of these animals contains 16%~22% proteins. Muscle protein can be divided into
myofibrillar protein, sarcoplasmic protein and matrix protein. Proteins of these three
170 Hong Lin, Lisha Wu and Shuhui Wang

categories differ markedly in their solubility. Sarcoplasmic proteins can be extracted by water
or buffers with low ionic strength (0.15mol/L or lower), myofibrillar protein can be extracted
only in high salt concentrations, and matrix proteins are insoluble. Sarcoplasmic proteins
contain large amounts of glycolytic enzymes and many other enzymes in addition to
myoglobin and haemoglobin, which affect the color of meat.
Myofibrillar proteins account for about half of animal skeletal muscle proteins. These
proteins are intensively charged hydrated and are insoluble under physiological conditions.
Matrix proteins participate in the formation of connective tissues. Collagen, reticulum
and elastin are examples of matrix proteins. Collagen is fibrous and occurs in all muscle
tissues. Reticulin refers to a type of fiber in connective tissues composed of type III collagen.
The proteins crosslink and form a fine mesh network. This network acts as a supporting mesh
in soft tissues such as liver, bone marrow, and the tissues and organs of the lymphatic system.
Elastic fiber is composed of the protein fibrillin and elastin made of simple amino acids such
as glycine, valine, alanine, and proline. Elastin helps skin to return to its original position
when it is poked or pinched. Elastin is also an important load-bearing tissue in the bodies of
vertebrates and used in places where mechanical energy is required to be stored. These matrix
proteins are less water soluble than sarcoplasmic and myofibrillar proteins.

6.2. Casein

Milk contains multiple types of proteins and the proteins exhibit different properties
(Table 5-11). In skimmed milk, casein accounts for about 80% of total protein. Casein is a
mixture of phospho-contaning proteins and precipitate out from skimmed milk at pH4.6 and
20°C. Casein is extremely hydrophobic and occurs as micelles in milk. Casein is composed of
αs1-casein, casein, β-casein, αs2-casein and κ-casein. The diameters of casein micelles range
from 50nm to 300nm with molecular weight range of 107~3×1010 Daltons. Casein micelles
are aggregated by sub-micelles, which has a diameter range from 10 to 20nm and molecular
weight range from 3×105 to 6×105.
Sub-micelles consist of αs1-casein, β-casein, αs2-casein and κ-casein and the fractions
aggregate through hydrophobic interaction. The core of sub-micelles is hydrophobic in
contrast to the hydrophilic surface. The carbohydrate-rich κ-casein is restricted on a region of
the surface through self-association. Therefore, the surface of sub-micelles contains one
carbohydrate-rich regions and one phosphate-rich region that is formed by other caseins.
Casein micelles are the aggregation of sub-micelles. The electrostatic interaction
contributed by colloidal calcium phosphate facilitates the aggregation of sub-micelles. The
aggregation occurs between the negatively charged serine residue on casein and the
positively-charged colloidal calcium phosphate, which occurs in Ca9 (PO4)6 clusters. Because
κ-casein contains no phosphate groups, the electrostatic interaction occurs only between α-
casein and β-casein.
During casein micelle formation, sub-micelles with low content of κ-casein or without κ-
casein are located in the interior of casein micelle. When the surface of the micelle is
completely covered by κ-casein, the micelles stop growing up.
Proteins 171

6.3. Whey Protein

The main constitutients of whey proteins are β-lactoglobulin, α-lactoglobulin and serum
albumin in decreasing order in content, as shown in Table 5-9. These proteins are soluble at
pH 4.6 in their natural or denatured forms. In the manufacture of caseinate and country
cheese, these protein fractions remain in the whey. Hence, whey protein is a by-product of
cheese making.
Whey protein is soluble in a wide range of pH, temperature and ionic strength, pH4~5,
which is near the isoelectric point. Solubility is most important physicochemical and
functional property of natural whey protein. In addition, whey protein produces stable gel
when heated. This property together with its surface properties imparts whey protein with
important application in the food industry.

Table 5-11. Proteins in milk [21]

Protein Content in skimmed milk (g/L) Relative molecular weight


Casein
αs1-casein 12~15 22068~23724
αs2-casein 3~4 25230
β- casein 9~11 23944~24092
κ- casein 2~4 19007~19039
Whey protein
β-whey protein 2~4 18205~18363
α-whey protein 0.6~1.7 14147~14175
albumin 0.4 66267
Immunoglobulin
IgG1 0.3~0.6 153000~163000
IgG2 0.05~0.1 146000~154000
IgA 0.05~0.15 385000~417000
IgM 0.05~0.1 1000000
Secretory component 0.02~0.1 79000

6.4. Wheat Protein

According to solubility, wheat proteins are divided into albumins (water soluble),
globulins (soluble in 10% NaCl but insoluble in water), gliadin (soluble in 70%~90%
ethanol), and glutenin (insoluble in water or ethanol but soluble in acid or alkali solutions).
Albumin and globulin account for about 10% to 15% of wheat endosperm proteins. The
two fractions contain free sulfhydryl and higher proportion of alkaline and charged amino
acids. Albumins are relative smaller molecules with molecular weight ranging from 12000 to
26000 compared with that of globulin of up to 100000. Gliadin and glutenin are the major
constituents of gluten and account for about 85% of flour protein. Both gliadin and glutenin
are complex mixtures of multiple protein components and the two fractions share similar
contents in wheat flour. Amino acid analysis indicates that gliadin and glutenin have very
high contents of glutamine and praline and very low contents of lysine and ionized amino
acids. Gliadin and glutenin are among the least charged proteins. Although the sulfur-
contaning amino acids are low, these amino acids play an important role in gluten structure
and dough formation.
172 Hong Lin, Lisha Wu and Shuhui Wang

The compact spherical structure of gliadin is related to the intramolecular disulfide bond.
Glutenins associate with each other through inter-molecuar disulfide bonding. Hence,
glutenins are found as large and stretched associated molecules. Glutenin is insoluble due to
intermolecular disulfide bonds, but treatment with reducing agents increases its solubility.
The formation of gluten is the result of both covalent and non-covalent interactions of
multiple protein constituents. Glutenins contribute to the formation of the network structure,
while gliadin and other proteins are trapped within the structure. Wheat flour contains 12%
protein, 70% starch and 2% fat. During kneading, these components are incorporated into the
gluten network structure and a starch-protein-fat composite matrix is formed.
The reduction of disulfide bonds during kneading the re-oxidation of sulfhydryl during
dough storage occur mainly in cross-linked glutenin. The reduction of disulfide bonds
facilitates the gluten relaxing and enhances the non-covalent interactions between fat, starch
and other additives, leading to the formation of a continuous starch-protein-fat complex.

6.5. Soybean Protein

Soybean contains 42% protein, 20% oil and 35% carbohydrates on dry basis. Soybean
protein is very sensitive to physical and chemical treatment. For example, heating under
moisture conditions and alteration of pH can cause significant changes in the physical
properties of soybean protein, such as solubility, viscosity and molecular weight.
Globulins are the most important constituents of soybean protein. Globulins are
insoluble in their isoelectric points, but the addition of NaCl or CaCl2 dissolves the proteins.
Globulins are soluble in water in the absence of NaCl or CaCl2 in other pHs than their
isoelectric points. Soybean proteins are least soluble in the pH range of 3.75~5.25, but are
most soluble in pH1.5~2.5 and that higher than 6.3. At pH6.5, about 85% nitrogen of defatted
soybean protein can be extracted by water and the yield can be further increased by 5%~10%
by increasing the pH.
According to the sedimentation coefficient, the water-extractable components of soybean
protein are divided into four fractions: 2S, 7S, 11S, and 15S, of which, 7S and 11S protein are
the most important. 7S protein accounts for 37% of the total protein, while the 11S fraction
accounts for 3l%. Soybean protein concentrates and soybean protein isolates are the most
important commercial products of soybean protein and their contents are higher than 70% and
90% respectively.
Soy protein concentrate can be prepared from defatted soybean meal in one of three
methods, namely by extraction with 60%~80% ethanol, with pH 4.5 aqueous solution, or with
water after heat denaturation. In these methods, extraction with the solvents remove soluble
carbohydrates from the material and the nitrogen content in the residue is increased as a
result. Removal of soluble carbohydrates markedly improves the flavor of soy protein
concentrate and flatulence factors are removed at the same time. Soy protein concentrates are
often used as the material of thermoplastic extrusion.
In the preparation of soybean protein isolate, proteins in defatted soybean meal are firstly
extracted with water at neutral or higher pH. Then, the proteins are precipitated by adjusting
the pH to the isoelectric point. The protein precipitate is collected and redissolved by
changing the pH and finally is applied to spray dryer. Soy protein isolate can be modified by
chemical, heat or enzymatic treatments to improve its functional properties.
Proteins 173

6.6. New Protein Resources Development

Due to the increase of world population and people‘s living standard, the demand on
protein keeps increasing and the deficiency of protein resources has turned to be a worldwide
issue. Improving the utilization efficiency of existing protein resources and exploiting new
protein resources are effective measures to solve the protein resource deficiency problem.

6.6.1. Proteins From Oil Crops


The byproducts of the oil processing industry are often used as feed or fertilizer. These
byproducts have high protein content and are good protein resources for human beings.
Soybean protein is a good example. Soybean contains about 40% protein and defatted soy
protein contains up to 50% protein. The contents of all the essential amino acids, except Met
and Cys, are close to the recommendations of FAO and their nutritional values are similar to
that of animal proteins. Hence, soybean protein is good vegetable protein.
At present, soybean protein is the most extensively used oil crop protein. Soybean
protein isolate has gained wide applications in noodles, bakery foods and many other foods
and the demand keeps increasing in recent years. It is expected that the yield and consumption
of soy protein powder will exceed those of soybean protein isolate after its production and
deodorization technologies are mature. Soy protein can significantly improve the quality of
flour-based foods. For example, it can whiten flour and replace existing chemical brighteners;
be added into noodles or dumplings to improve their tenacity and reduce the dissolution rates
of starch during cooking; be added to bakery foods to improve biscuit crisp, bread tenacity,
and cake softness; be added to buns, steamed buns and other steaming food to yield smooth
surface; and be added to instant noodles and other fried foods to reduce oil consumption and
the greasy mouth feel of fried foods.

6.6.2. Single Cell Protein


Single-cell protein refers to such micro-organisms as yeast, lactic acid bacteria and mold,
etc. which are cultivated by industrial means. This biomass is rich in protein, which can be
used as human food or animal feed. Single-cell proteins are concentrated protein products and
their crude protein contents range from 50% to 85%. Single cell proteins have complete
amino acid composition and their bioavailabilities are very high. In addition to proteins, the
biomass also contains vitamins, inorganic salts, fats and sugars. Hence, single cell proteins
have higher nutritive values than fish powder and soybean powder. Besides, single cell
proteins have the following advantages in protein resource development.

1) A wide range of materials can be used for single cell protein production. For example,
fiber resources such as straw, wood chips and bagasse can be used as the fermentation
substrate.
2) The investment for single cell protein production is low and the biomass grows very
quickly. The generation time of microorganisms is about only tens of minutes. It is estimated
that a 500 kg cow produces only 0.4kg protein, while yeast of the same weight can produce at
least 5000 kg proteins per day.
3) The industrial production of single cell protein does not need large area of arable lands
and are not affected by seasons and climate conditions.
174 Hong Lin, Lisha Wu and Shuhui Wang

The development of single cell protein resources has bright prospect. However, the high
content of nucleic acid in most single-cell proteins restricts their direct consumption by
consumers. Heat and alkaline treatment can improve the digestibility and amino acid
availability of single cell protein and reduce nucleic acids contents. Long-term toxicological
study indicates that heat- or alkaline-treated single cell protein is nontoxic.

6.6.3. Insect Protein


Insects have the largest biotic population on the earth and are characterized by high food
conversion rate, rapid propagation and high protein contents. Insects have been recognized as
the largest and most potential animal protein resource in the world. The protein contents of
many insects exceed 50% on dry basis. For instance, the protein contents of maggot butterfly,
crickets, cicadas and ants are 61%, 71%, 75%, 75%, 72% and 67% respectively. In some
species, the protein contents are even greater than 80%. The protein contents of these insects
are far greater than those of chicken, fish, pork and eggs, and is similar to that of beef liver.
Meanwhile, the distribution and ratio of essential amino acids in insect protein is close to the
ratios recommended by FAO. Therefore, insects can provide high-quality proteins.

6.6.4. Leaf Protein


Leaf proteins are extracted from the green stems and leaves of plants. After the materials
are squeezed, the juice is collected and the proteins are then separated, concentrated, and
dried. Leaf protein products contain 55%~72% protein and 18 amino acids, including 8
essential amino acids. The amino acid composition is close to the recommendations of FAO
for adults and the Lys content is high. Hence, leaf protein is potential for consumers of the
Third World countries in which cereals are consumed as the staple food. Leaf proteins have
high contents of Ca, P, Mg, Fe and Zn, which are 5~8 times higher than seeds. Besides, the
contents of carotenes and lutein are 20~30 and 4~5 times higher than those of leaves
respectively. Lead proteins do not contain cholesterol as in animal proteins and have multiple
physiological functions. Leaf protein has been recognized by FAO as a high-quality protein
source and is a potential protein resource.

7. NUTRITION AND SAFETY ASPECTS OF PROTEIN


The nutritive values of proteins vary with their sources and the contents of essential
amino acids and their digestibility are the main factors that determine proteins‘ nutritive
value. Therefore, the daily protein requirement depends on the quality and quantity of
proteins in foods.

7.1. Quality of Protein

The quality of proteins is mainly decided by the composition of essential amino acids
and their digestibility. High-quality proteins should contain all the essential amino acid and
their contents should be higher than the reference levels recommended by FAO/WHO/UNU.
Besides, the digestibility of high-quality proteins should be comparable or greater than that of
albumen and milk protein.
Table 5-12. Essential amino acid contents and nutritive values of proteins derived from several foods
(mg/g protein) [18, 19]

Item Source
Egg Milk Beef Fish Wheat Rice Corn Barley Soybean Broad bean Pea Peanut Kidney
bean
His 22 27 34 35 21 21 27 20 30 26 26 27 30
Ile 54 47 48 48 34 40 34 35 51 41 41 40 45
Leu 86 95 81 77 69 77 127 67 82 71 70 74 78
Lys 70 78 89 91 23(1) 34(1) 25(1) 32(1) 68 63 71 39(1) 65
Met+Cys 57 33 40 40 36 49 41 37 33 22(2) 24(2) 32 26
Phe+Tyr 93 102 80 76 77 94 85 79 95 69 76 100 83
Thr 47 44 46 46 28 34 32(2) 29(2) 41 33 36 29(2) 40
Trp 17 14 12 11 10 11 6(2) 11 14 8(1) 9(1) 11 11
Val 66 64 50 61 38 54 45 46 52 46 41 48 52
Total 512 504 480 485 336 414 422 356 466 379 394 400 430
Protein content (%) 12 3.5 18 19 12 7.5 / / 40 32 28 30 30
Chemical score /% (by 100 100 100 100 40 59 43 55 100 73 82 67 /
FAO/WHO pattern)
PER 3.9 3.1 3.0 3.5 1.5 2.0 / / 2.3 / 2.65 / /
BV(on rat) 94 84 74 76 65 73 / / 73 / / / /
NPU 94 82 67 79 40 70 / / 61 / / / /
Note: Chemical score is defined as the proportion of the content of a limiting amino acid in the material to that of the reference protein.
176 Hong Lin, Lisha Wu and Shuhui Wang

Table 5-13. Recommended patterns of essential amino Acids in foods


(mg/g protein) [19]

Amino Recommended pattern Amino acid Recommended pattern


acid Infant Preschool Adults Infant Preschool adults
(2~5) children (2~5) children
His 26 19 16 Phe+Tyr 72 22 19
Ile 46 28 13 Thr 43 28 9
Leu 93 44 19 Try 17 9 5
Lys 66 44 16 Val 55 25 13
Met+Cys 42 22 17 Total 460 241 127

The proteins derived from major cereals and beans are deficient in at least one essential
amino acid. For example, the proteins of rice, wheat, barley and oat, are deficient in Lys but
rich in Met and this is reversed for the proteins of beans and seeds of oil crops. The proteins
of some seed plant seeds, such as peanuts, are deficient in both Met and Lys. Essential amino
acids with contents lower than that in reference protein are defined as limiting amino acids.
Adults that intake proteins only from crops and beans cannot maintain health and children
that intake only one of the two kinds of protein mentioned above cannot grow in the normal
rate. Table 5-12 lists the contents of essential amino acid in some proteins.
Generally, the proteins derived from plants and animals contain enough His, Ile, Leu,
Phe, Trp and Val and these amino acids are often not limiting amino acids in common foods.
However, Lys, Thr, Trp and sulfur-containing amino acid are often present in insufficient
quantity and are regarded as limiting amino acids.
Mixing a protein that deficient in an essential amino acid with another protein that is rich
in the essential amino acid can increase the nutrition value of the protein. For example, the
mixture of cereal protein and bean protein can provide complete and balanced essential amino
acids. The nutritive value of proteins can be improved also by supplementing the
corresponding limiting amino acids. For instance, supplementation of Met and Lys improves
the nutritive value of cereal and bean proteins.
If the protein or protein mixture contains all the essential amino acids and the contents
and proportions of the amino acids can maintain the optimum growth rate and health
conditions for consumers, the protein or protein mixture is though to have an ideal nutritive
quality. Table 5-13 lists the ideal amino acid distribution patterns for adults and children.

7.2. Digestibility

Protein digestibility is defined as the proportion of absorbed nitrogen to ingested


nitrogen. The contents of essential amino acids in proteins are important indicators of protein
quality. However, the utilization of these amino acids determines the real quality of proteins.
Table 5-14 lists the digestibility of various foods proteins in human body. It could be seen
that plant proteins have higher digestibility than animal proteins.
The digestibility of proteins is affected by the following factors:

(1) Structure: The structure of proteins markedly affects their hydrolysis by enzymes.
Generally, native proteins are more resistant to enzymatic hydrolysis than their denatured
forms and the hydrolysis of insoluble fibrous proteins is more difficult than that of thoroughly
denatured globular proteins.
Proteins 177

Table 5-14. Digestibility of some food proteins in human body [19]

Protein source Digestibility (%) Protein source Digestibility (%)


Egg 97 Pea 88
Milk, cheese 95 Peanut 94
Meat, fish 94 Soybean meal 86
Corn 85 Soybean protein isolate 95
Refined rice 88 Kidney bean 78
Whole wheat 86 Corn products 70
Refined flour 96 Wheat products 77
Gluten 99 Rice products 75
Oat 86 Wheat 79

(2) Antinutrients: Most vegetable proteins isolated and protein concentrates contain
trypsin inhibitors, chymotrypsin inhibitor, and exogenous lectins. The inhibitors prevent the
complete hydrolysis of proteins derived from legumes and oil seeds by trypsin. Lectins hinder
the absorption of amino acids in intestines. Heat treatment can destroy these antinutrients and
make proteins more susceptible to digestion. In addition to the antinutrients mentioned above,
proteins derived from plants also contain phytic acid, tannin and other anti-nutrients.
(3) Association with other components: Association with polysaccharides and dietary
fiber reduces the hydrolysis speed and thoroughness of proteins.
(4) Processing: Heat and alkaline treatment can cause chemical reactions in proteins,
such as Lys residue generation. These changes reduce the digestibility of proteins. Besides,
Millard reactions might occur between reducing sugars and proteins. These reactions reduce
the digestibility of Lys residues.

7.3. Toxic Proteins

Toxic proteins have been found in many kinds of plants. Chemically, the toxic proteins
can be divided into two categories: lectins and enzyme inhibitors. Lectins occur widely in
leguminous plants, such as soybeans, peas, kidney beans, sword bean and castor seed. These
toxins can agglutinate red blood cells. Hence, the intake of uncooked or undercooked seeds of
these plants can cause poisoning. The main symptoms of lectin poisoning include nausea,
vomiting, and even death. Enzyme inhibitors are found in beans, potatoes, taros, wheat and
immature bananas. These proteins inhibit the activities of trypsin or amylase and affect the
digestion and absorption of nutrients by human body.

7.3.1. Toxic Peptides


Toxic peptides are found in the most quantity in poisonous weeds. Amanita mushroom
and phallus mushroom contain the highest levels of poisonous peptides. Intake of as low as
50 grams of these mushrooms could lead to death.

7.3.2. Toxic Amino Acids


Some legumes contain toxic amino acids. For example, grass vetch contain α,γ-2-amino
butyric acid and kidney bean contains β-cyano-alanine. Canavanine in canavalia hampers
arginine metabolism in the body and L-3, 4-dihydroxyphenylalanine in kidney bean can cause
acute hemolytic anemia. Consumption of excessive green kidney bean can lead to poisoning
no matter whether the whether the beans are cooked or peeled.
178 Hong Lin, Lisha Wu and Shuhui Wang

7.3.3. Allergic Proteins


In most cases, food-induced irritability is caused by the proteins in foods. For example,
the parvalbumins in myogen of fish are sensitinogens. Parvalbumins are small acidic
glycoproteins with molecular weight of 11-12kD. The isoelectric point of these proteins are
around 4.75 and two Ca2+ ions are bound to the Asp-Asp-Ser-Glu-Glu-Phe and Asp-Asp-Asp-
Glu-Lys regions. Allergic proteins in crustacean are mainly tropomyosins, of which, an acidic
glycoprotein with molecular weight of 36kD, isoelectric point of 4.5, and sugar content of
4.0%, has been identified as the major sensitinogen. With the development of technologies,
more and more allergic proteins will be identified from crustacean. At least 13 allergic
proteins have been identified in crustacean. The molecular weights of these proteins are in the
range 16~166kD and most are in the molecular weight 36kD. These proteins do not contain
Trp and have low Tyr and Phe contents.

8. CHANGES OF PROTEINS DURING FOOD


PROCESSING AND STORAGE
8.1. Effects of Processing Methods on Protein Quality

During food processing, temperature, moisture, pH, presence of other components and
pressure are main factors that lead to protein changes. Among these factors, moisture and
temperature have the largest effect on proteins. Hence, only the effects of dry and moist heat
treatments on protein are detailed.

8.1.1. Boiling
Boiling is a common processing method in the food industry. Boiling does not reduce
the quantity of proteins or essential amino acids of fish and meat. Boiling of milk neither
affects the amino acid content nor reduces the bioavailability. The egg white and yolk have
different sensitivity of heat treatment. The digestibility of raw egg white is only 50%, but raw
yolk can be digested and absorbed completely. Moderate cooking of eggs can yield of nearly
100% of digestibility and biological value due to heat denaturation of proteins. Moist heat
treatment is desired for most vegetable proteins, especially for proteins derived from beans.
Moist heat treatment increases the nutritive value of bean proteins due to protein denaturation,
inactivation of protease inhibitors and toxic substances, and disrupt of tissues and starch
granules.
Enzymes, such as lipase, lipoxygenase, protease, and polyphenol oxidase, can be
inactivated during blanching or steaming. Enzyme inactivation can prevent the formation of
undesired colors and flavors and the loss of vitamins. Heat treatment of rape seeds inactivates
myrosinase and avoids the formation the goitrogenic 5-vinyl-2-sulfur-oxazolidinone.

8.1.2. Pasteurization
It is generally believed that either the tank pasteurization method or the high-temperature
short time pasteurization has only slight effects on milk proteins, though whey proteins are
partially denatured. Extensive studies on infants, school-age children, adults and experimental
animals indicate that the nutritional value of milk is not affected by pasteurization.
Sealed container must be heated thoroughly until the contents are sterile. The required
intensity of heat treatment depends largely on the physical state of the contents. In
Proteins 179

pasteurization, the heat transferred from the vessel wall to the center in semi-solid foods is
much less than in foods containing liquids as the heat transfer medium. High-frequency
oscillatory electron heating and aseptic canning can overcome these problems, but their
applications are still very limited.
Pasteurization reduces the biological value of liquid milk by 6%, lysine content by 10%
and cystine content by 13%. The decrease in biological value may be due to the loss of
cystine. Pasteurization has much more significant effects on concentrated milk. Hence,
pasteurization can significantly reduce the bioavailability of proteins with limited lysine
contents.

8.1.3. Can Sterilization


Meat cans need more intensive sterilization conditions than dietary products, because the
heat transfer in solids or semi-soldiers are more difficult than in liquids. The heat sterilization
method used in the current meat industry does not lead to significant loss of amino acids,
except cysteine. However, canned beef has reduced digestibility and biological value than
unprocessed beef. Besides, the bioavailability of Lys and Met in canned meats is slightly
lower than that of fresh meat. Similar trends have also been found in canned fish.
Sterilization reduces the biological values of most animal proteins. In contrast, the
biological values of many vegetable proteins are increased after suitable sterilization. This is
because sterilization not only improves the digestibility, but also destroys natural anti-
nutrients. In the heat treatment of soybean products, both the loss of nutrients and inactivation
of toxic components (such as typsin inhibitors) must be taken into consideration. The
optimum sterilization conditions should minimize nutrient loss and maximize toxicant
inactivation. Researches on moist heat treatment-caused Lys loss indicate a first-order
reaction with reaction rate constant of 0.166/h and activation energy of 125.5kJ/mol.

8.1.4. Dehydration
Milk and dairy products can be dehydrated by spray drying or drum drying. The effects
of these methods on the nutritional value of milk proteins have attracted wide attentions. In
drum drying, milk is spread on the drum and the nutrient loss varies with the conditions
applied. Dehydration might lead to the loss of Lys. However, since Lys is not the limiting
amino acid of milk, moderate dehydration dose not impair the nutritive value of milk powder.
At present, dehydration has not been widely used in the industrial processing of meats,
because dehydration cannot impart meats with competitive price, quality, and consumer
acceptability compared with other products, through freeze-dried meats are more nutritional
than heat-treated or irradiated products. Nevertheless, the quality of freeze-dried products
cannot last long and critical changes might occur during storage.
Fish meal prepared from fish by dehydration is widely used as feed and only very few
are used in human foods. Fish can be dehydrated in many technologies, including flame
drying, steam pipe drying, and solvent drying. Therefore, the changes of protein quality vary
with the technology used.

8.1.5. Freeze-Drying
Most foods are dried in conventional methods. However, freeze-drying has potential
applications in the drying of unstable and gourmet foods. Studies have confirmed that freeze-
drying under proper conditions does not impair the quality of proteins.
180 Hong Lin, Lisha Wu and Shuhui Wang

8.1.6. Baking and Puffing


Cereal foods are often dried by dry heating. During dry heating, the moisture contents in
materials decrease rapidly and proteins might be lost seriously. Baking causes a marked loss
of the nutritional value of proteins on material surface. Hence, cookies and biscuits suffer
serious protein loss during processing and the losses depend on the thickness of the foods.
Generally, the loss of bioavailable amino acids in biscuits are is the function of thickness,
heat intensity and heating time. The presence of sucrose increases the loss of bioavailable
Lys. Breads lose 10%~15% of lysine during baking, and further 5% of lysine is lost when
breads become hard.
Intensive baking or puffing can lose proteins nutritive value in instant cereal foods.
Though the content of Lys is only slightly affected, its bioavailability is significantly reduced
after baking or puffing.

8.1.7. Other Processing


The application of various protein isolates in the food industry keeps increasing in recent
years. In the production of these isolates, the oilseed protein and fish protein are extracted
with alkali; then, the protein in the supernatant is precipitated by acid or heat treatment to
various yield protein isolates. These products can be used directly in foods or after medication
by spinning. Treatment of casein, peanut flour and soybean protein with strong alkaline
reduces the quality of the proteins and generate new amino acids, such as lysinoalanine,
which is the condensation compound of lysine and alanine.

8.2. Mechanisms of Protein Loss in Food Processing

8.2.1. Effects of Heating


Heating affects the spatial arrangement of food proteins and enhances the thermal
oscillation of the molecules. These changes weaken the inter-molecule interaction, unfold the
proteins, and disrupt the disulfide bridges. This process is termed heat denaturation. Heat
denaturation only changes the structures of proteins and cannot alter amino acid sequences.
The denaturation induced by moderate heat treatment is reversible. However, the denaturation
of most proteins occurred during food processing is irreversible.
Denatured proteins have different solubility and functional properties compared with
their natural forms. Hence, protein denaturation plays important role in food processing.
However, the nutrition aspects of protein denaturation have attracted much less attention. It is
generally believed that denatured proteins are more susceptible to enzyme hydrolysis and thus
have improved nutritive values.
In digestion, proteins are firstly denatured and hydrolyzed by the combination of
hydrochloride acid and pepsin in the stomach. Heat denaturation facilitates this process and is
hence believed to be beneficial from the point of nutrition view. However, this is not always
the case. If a protein plays a special function in the digestive tract, it is crucial to maintain its
structure after ingested. For example, it has been proved that feeding of denatured whey
protein is not good for the growth of calves. This truth has also been found in newborn
infants. Proteins in breast milk are only partially digested and these proteins are more
resistant to trypsin hydrolysis compared with milk proteins. It is therefore believed that some
proteins have specific functions in the digestive tract. Some of such proteins have been
identified. For example, lactoferrin is involved in iron delivery; immunoglobulin imparts
Proteins 181

Infants with passive immunity; lysozyme destroys microorganisms; and streptogenin


promotes the growth of lactic acid bacteria.

8.2.2. Effects of other Components

(1) Protein-carbohydrate interactions

Moderate reactions with reducing sugars (Maillard reactions at early stage)

Maillard reactions, also termed non-enzymatic browning reactions, occur between


reducing sugars and the free amino groups of peptides, proteins, and amino acids. Carbonyl
compounds, such as aldehydes and ketones, also undergo Maillard reaction with reducing
sugars. Maillard reaction occurs readily during the processing and storage of foods, especially
in dietary products. The rates of Maillard reactions depend on the temperature and moisture
of foods.
The first step of Maillard reactions includes the condensation between free amino groups
and carbonyl groups. The condensation products are converted to Schiff's bases, which are
subsequently isomerized to N-substituted glycosylamines. N-substituted glycosylamines
derived from amine compounds are relative stable, and those derived from amino acids are
unstable and immediately undergo Amado‘s rearrangement to produce 1-amino-l-deoxy-2-
ketones, which are the major products of Maillard reaction in milk after mild heating.
The ketosamines are subject to further reactions and the products include reductones,
furfural and unsaturated carbonyl compounds. These compounds then react with each other to
produce soluble polymers and finally insoluble melanoidins in sequence. All the Lys residues
and amino acids at the N-terminal are completely destroyed in these reactions.
In addition to nutritional value loss of proteins, Maillard reactions can produce potential
harmful compounds. It has been proved that the brown soluble compounds (pre-melanoidins)
generated by the thermal reaction between free amino acids and glucose have particular
physiological and anti-nutritional properties. However, no physiological functions have been
found in pre-melanoidins formed by proteins and sugars, because the addition of severely
heated protein-glucose mixture to a well balanced diet dose not affect the growth function of
experimental animals.

Violent reaction with carbonyl groups (Maillard reactions at later stage)

Many researchers have confirmed that reactions of proteins with glucose at high
temperature and pressure significantly reduce the nutritive value of proteins, even when the
reaction time is short. Under these conditions, carbonyl compounds cause extensive cross-
linking of proteins through Strecker degradation and aldol condensation. In this case, the
value of any protein as the amino acid source is very limited.

Reaction with sucrose

As mentioned above, reducing sugars can react with proteins and cause amino acid loss.
However, it has also been found that sucrose is also involved in the reaction with proteins.
The initial reaction between lysine and sucrose is the 'aminolysis' of the 1, 2 glycosidic
linkage between the glucose and fructose residues by the ε-amino group of lysine. Breakdown
182 Hong Lin, Lisha Wu and Shuhui Wang

of the glycosidic linkage yields reducing groups, which undergo typical Maillard reaction
with another molecule of lysine. It has been reported that sucrose at relatively high levels in
oil seeds may be largely responsible for the damage to protein quality observed when they are
severely processed [22].

(2) Protein-lipoid interactions


The reactions between oxidized lipids and proteins have been extensively reported.
However, no general conclusions can be drawn on the reactions, because lipids are oxidized
in multiple modes. During oxidation, unsaturated fatty acids firstly bind to catalysts to
produce free radicals. Free radicals then react with oxygen to form hydrogen peroxides,
which can undergo reactions in a variety of ways. Some of the products are carbonyl
compounds that can participate in Maillard reactions. These products are subject to either
degradation or polymerization.
When oxidized lipids and proteins are incubated, cross-linking occurs between lipid free
radicals and proteins, which reduces the protein solubility, increases the resistance to
digestive enzymes, and destroy most amino acids. Then, sulfur-containing amino acids are
oxidized by hydroperoxides and methionine is converted to methionine sulfoxide. Free
methionine sulfoxide as a methionine source is the same as methionine. However, methionine
sulfoxide in peptides or proteins is less bioavailable than methionine, because it is resistant to
enzymatic hydrolysis. Cystine is quickly degraded and cysteic acid is one of the degradation
products. Cysteic acid is not a source of cystine for human body.
The newly formed active carbonyl compounds can react with the amino group of Lys
resides in a way similar to browning reactions. These reactions make Lys physiologically
unavailable.
In electrostatic fields, oxidized lipids can react with amino group-free proteins to yield
simple compounds. In this case, the nutritive loss of proteins is very small.

(3) Protein-aldehyde interactions.


In addition to the interactions between carbonyl and amino groups mentioned above, two
special reactions of proteins with aldehydes have also been identified during food processing.
One is the reaction with gossypol during cottonseed processing. The other is the reaction with
formaldehyde during meat fumigation. Both the two reactions decrease the bioavailability of
lysine.

8.3. Effects of Processing on the Nutritional Value of Proteins

Proteins as food materials are exposed to physical or chemical treatments during


blending and application. These treatments affect the functional and nutritional properties of
proteins. Besides, purposive modifications have been applied to improve the existing
properties of protein or create new functions.

8.3.1. Heat Treatment


Heat treatment can cause protein structure change, peptide bond hydrolysis, amino acid
side chain modification and condensation with other molecules. The later two reactions are
harmful to the nutritive value of proteins. Structural changes and limited peptide bond
hydrolysis caused by mild heat treatment do not affect the nutritive quality of proteins, but
Proteins 183

significantly affect the functional properties of proteins. The degree and result
(conformational change and aggregation) of thermal denaturation depend largely on the
nature of proteins and environmental conditions. Mammalian collagens start to unfold,
dissociate, and dissolve when heated to 65°C in the presence of large quantity of water. In
contrast, actins undergo contraction, aggregation and water holding capacity reduction under
same conditions.
Caseinate monomers are very resistant to heat treatment. Vigorous heat treatment dose
not affect the properties of caseinate. Mild heat treatment dose not lead to the destruction or
generation of covalent bonds or serious changes of higher structures in proteins. Hence, from
the nutritional point of view, the changes induced by mild heat treatment are beneficial.
Heat treatment can eliminate the toxic effects of lectins in legumes, because the lectin is
a thermal unstable protein which can bind polysaccharides. It can reduce the nutritional value
of natural plant proteins in the diet. The presence of lectins can form compounds by
combining intestinal brush border membrane polysaccharide, which can diminish the abilities
of shifting and digestion in amino acid, produce toxicity to human and animal; heat treatment
can eliminate their harmful effects.
Protease inhibitors in foods, including trypsin inhibitors and chymotrypsin inhibitors, are
inactivated during heat treatment. High temperature treatments, such as high-pressure
sterilization, puffing, sterilization, cooking and baking, can destroy all antinutrients factors
(Figure 5-14). Therefore, moderate heat treatment can markedly improve the nutritional value
of vegetable proteins.
Many proteins, such as soybean globulin, collagen and egg albumin, are more digestible
after moderate heat treatment.

Figure 5-14. Effect of steaming on trypsin inhibitor activity and protein efficiency ratio of soybean
meal feed [20]

After heat treatment, the proteins are more susceptible to enzymatic hydrolyze due to
unfolding and amino acid residues exposure. Besides, heat treatment can remove off-odor
compounds in soybean protein and avoid the undesired flavor caused by lipid oxidase.

8.3.2. Dehydration
When the water in a protein solution is removed, aggregation occurs to the proteins. This
is the case especially when water is removed by evaporation at high temperatures, because the
184 Hong Lin, Lisha Wu and Shuhui Wang

solubility and surface activity decline sharply. Hence, the effects of drying on the functional
properties of protein should not be ignored. Drying changes the size and internal and surface
porosity of particles and consequently alters the wettability, absorption, diapersion and
dissolution of food proteins. When water is quickly removed as steam, particles shrink in the
minimum level and salts and carbohydrates migrate to the dried surface, yielding porous
powder particles. This change can be observed in freeze-drying and spray-drying.
Introduction of bubbles to protein solution before drying or the implementation of controlled
aggregation during drying can be applied to increase the porosity of particles.

8.3.3. Irradiation
Irritation causes inter- and intra-molecular cross-linking of proteins or peptide bond
breakdown in foods with low moisture content. In the presence of catalase, Tyr is oxidized
and condensed to dityrosine.

8.3.4. Oxidation
Oxidants are important additives in the food industry. For example, hydrogen peroxide is
sterilization and bleaching agent which can be used as a low-temperature fungicide in
package. It can also been used to improve the color of fish protein concentrates, flours and oil
seed protein isolates. Benzoyl peroxide can oxidize cysteine in flour to cystine so as to
enhance gluten strength and bleach flour. In some cases, benzoyl peroxide can also be used in
whey powder bleaching. Peroxides are also found in foods without artificially added oxidants
and these peroxides are often the reason for the degradation of proteins present in the foods.
Light oxidation, irradiation, trace metals, oxygen, hot air drying and aeration during
fermentation can all induce oxidative denaturation of amino acids in proteins.
Trp is the most sensitive amino acids to oxidation, followed by tyrosine and histidine in
sequence. The oxidation of sulfur-containing amino acids occurs mainly in the sulfur-
containing side chains and that of aromatic-containing amino acids mainly occurs in the side
chains containing aromatic rings.

8.3.5. Maillard Reaction


Foods containing proteins and reducing carbohydrates or carbonyl compounds (such as
aldehydes and ketones produced by lipid oxidation) are subject to Maillard reactions during
processing and storage. These reactions yield brown or black melanoidins, which impart
breads and bakery products with the brown color. The formation of melanoidins is
accompanied by the cross-linking of a small amount of proteins. The cross-linking
significantly reduces the digestibility of these proteins. Studies on some protein-carbohydrate
model systems reveal that melanoidins are mutagenic and the capacity depends on the degree
of Maillard reactions. Melanoidins are insoluble in water and are only weakly absorbed on
intestinal wall. Hence, melanoidins are only slightly physiologically toxic to human.
However, the low molecular weight precursors of melanoidin are absorbed easily.

8.3.6. Enzyme Treatment


Enzymatic hydrolysis can improve the quality of food proteins. For example, papain can
tenderize meat and chymotrypsin can coagulate casein. Papain, bromelain, pepsin and
protease produced by microorganisms can partially hydrolyze thermal denatured or solvent
denatured proteins and improve their solubility. The hydrolytes can be used in acidic or hot-
Proteins 185

processed drinks. The hydrolytes can also be used as alternative protein sources for patients
allergic to milk and gluten or those with digestive tract diseases.
Partial hydrolysis can improve the emulsifying and foaming abilities of thermally
denatured proteins, because the increased solubility of proteins facilitates their diffusion to
the oil/water and gas/water interfaces. However, when the degree of hydrolysis exceeds
3%~5%, the viscosity and thickness of protein layer absorbed on the interface are insufficient
to stabilize emulsion and foam. Hydrolysis reduces the volume of proteins, which is
unfavorable for the gelling ability and viscoelasticity of proteins, but limited hydrolysis of
gluten can enhance the expansion degree of dough in bread making and improve the flakiness
of biscuits.
In addition, proteases can connect low molecular weight peptides through peptide
transferring to produce new larger peptides, namely proteinoids.

8.3.7. Functional Peptides from Protein Hydrolysates


Proteins often undergo hydrolysis during processing. Hydrolysis not only improves the
nutritional value of foods, but also yields functional peptides, such as those with
antihypertensive, immunomodulatory, neuroactive, antimicrobial and antioxidative activities.
Hydrolysis using proteases has been preferred as the main method for producing functional
peptides from food proteins. Fish and other seafood proteins have gained particular interest as
potential functional peptide sources among various food protein resources due to their
abundance as byproducts. For example, antioxidative peptide could be prepared from Alaska
pollack (Theragra chalcogramma) and yellowfin sole (Limanda aspera) frame proteins by
using a crude enzyme mixture from mackerel intestine [23, 24]. The pepsin hydrolysate,
which exhibits a strong DPPH scavenging activity at a concentration of 6.80μg/mL, could be
obtained from a marine fish half-fin anchovy [25] and its antibacterial activity is higher than
the hydrolysates produced by other proteases [26].
Functional peptides can be generated by protease hydrolysis and/or fermention. Use of
exogenous enzymes is preferred in most cases over autolysis. Various food-grade proteases,
such as alcalase, flavourzyme and protamex from microorganisms, papain from plant, and
pepsin and trypsin from animal sources, have been widely used in producing functional
peptides in large scale.
Some functional peptides can be generated during fermentation or food processing. In
fermented foods, antioxidative peptides can be produced by the action of microbes and
endogenous proteolytic enzymes. For example, the free radical scavenging peptides could be
produced from fermented mussel sauce [27]. Milk whey fermented by lactic acid bacteria has
been reported to contain antioxidative peptides [28]. The peptide cyclo (His-Pro) is an
endogenous cyclic dipeptide with antioxidant activity. It has been isolated from processed
foods such as dried shrimp and fish sauce [29].

9. CONTRIBUTION OF PROTEINS TO THE COLOR,


FLAVOR, AND TASTE OF FOODS
9.1. Bitterness
Some amino acids are bitter and the partial hydrolysis of proteins can also yield bitter
peptides, such as Leu-Leu and Arg-Pro. The bitterness of protein hydrolyzates and mature
cheese is contributed by short peptides and amino acids.
186 Hong Lin, Lisha Wu and Shuhui Wang

Peptides with molecular weight greater than 6kD cannot the action sites of taste
receptors and are hence tasteless. Whether a peptide with molecular weight less than 6kD is
bitter and its bitter intensity is related to its hydrophobicity. For details, see Section 5.1.3.

9.2. Off-Odors

Some protein products must undergo the deodorizaton process to remove off-odor
compounds bound to proteins. Ketones, aldehydes, alcohols, phenols, and lipid oxidation
products can produce beany flavor, rancidity, bitterness, and harsh taste. These compounds
can bind to proteins. When the proteins are cooked or chewed, these compounds are released
and the undesired tastes or odors are delivered to consumers, which markedly reduce the
acceptability of foods.
In addition to off-odors, proteins can also serve as the carrier of flavors. For example,
textured vegetable proteins can impart the meat flavor. Ideally, the desired volatile flavor
components should bind to carriers during storage and processing and be released rapidly and
completely in the oral cavity upon ingestion. Any factors that change the conformation of
proteins affect the binding to volatile compounds to proteins. In the presence of water
facilitates the binding to polar compounds to proteins, but shows no effect on that of nonpolar
compounds. Volatile compounds have only limited diffusion in dried protein components.
However, when the water activity is slightly improved, volatile compounds diffuse to their
binding sites rapidly. Casein can bind more volatile carbonyl compounds, alcohols and esters
in neutral and alkaline conditions than in acidic solutions. Oxidates and sulfates in high
concentrations can cause protein unfolding and thus improve the binding of proteins to
carbonyl compounds. Agents that can dissociate proteins or interrupt disulfide bonds improve
the binding of proteins to volatile compounds. When oligomers are dissociated to subunits,
the hydrophobic areas between subunits are masked during conformation change of the
subunits. Hence, the binding to nonvolatile compounds is reduced.
Thorough hydrolysis of proteins reduces their ability of binding volatile compounds. For
example, one kilogram of soybean protein can bind 6.7 mg caproaldehyde. After the protein
is hydrolyzed by acidic protease, the binding capability is reduced to 1 mg/kg protein. Hence,
the beany flavor of soybean protein can be alleviated by hydrolysis. Thermally denatured
proteins generally have improved volatile binding capabilities. For example, when 10%
soybean protein isolate is heated at 90°C for 1~24h in the presence of caproaldehyde and is
then free dried, the amount of caproaldehyde bound is 3~6 times more than that of unheated
protein. Dehydration process, such as free drying, can release more than 50% of bound
volative compounds. However, if a volatile compound has a low vapour pressure and is
present in low content, it can be well retained after the treatments. It should be noted that the
presence of lipids is beneficial for the binding and retention of volative compounds, including
those generated by lipid oxidation.

9.3. Sweetness of Protein Derivates

Development of safe and strong sweeteners by the modification of non-sugar and


unsweet natural compounds is an emerging field since 1960s. Such sweeteners that have been
put into industrial production are divided into tow categories: amino acids and their dipeptide
Proteins 187

derivates, and dihydrochalcone and their derivates. The potential of perilla aldehyde and its
derivates as sweetener has also attracted much attention.
Some naturally occurring amino acids have been found to be sweet, including Gly, D-
Ala, L-Ala, D-Ser, L-Ser, D-Thr, L-Thr, D-Trp, D-Pro, D-hydroxyproline, and D-Glu. Some
amino acid derivates are also found to be sweet. For example, the specific saccharinity of 6-
methyl-D-Trp reaches 1000 and is a potential novel sweetener.
In 1969, it was found that many dipeptide derivates of aspartic acid are sweet and
phenylalanine methyl ester (aspartame) has been approved by FDA for use in foods as a
sweetener. Aspartame is the sweetest compound among all sweeteners identified and is 2.7
times sweeter than sucrose. These dipeptide sweeteners are non-sugar compounds. Their
components are natural molecules and can be metabolized by human body. Besides, their
sweetness curves fit that of sucrose very well and the sweetness tastes very well. The
shortcoming is that they are unstable in high temperatures.

Sweet dipeptide derivatives have the following structural characteristics:


The two amino acids are of the L form.
The amino acid in the N terminal is Asp and its –COOH and –NH2 groups are free.
Another amino acid is neutral and one of the ends must be esterified. The smaller the
ester group, the sweeter the dipeptide.
The sweet intensity decreases with the increase of molecular weight.

9.4. Flavor Binding

The proteins of food have little flavor of their own, but are known to bind trap aroma
compounds. The most extensively studied protein is the milk protein β-lactoglobulin. The
protein can interact with many flavour compounds, such as aldehydes and ketones, ionones
and esters. Both reversible binding and irreversible binding can occur between proteins and
flavor compounds. The type of interaction between a protein and a flavor compound depends
on the nature of both the protein and the aroma compound. As most aroma compounds are
hydrophobic in nature, hydrophobic and reversible binding is predominant. On the other
hand, certain flavor compounds, such as aldehydes, can form covalent bonds with proteins
and such interactions are irreversible.
Proteins can be used as flavor carriers in the food industry. It is suggested that β-
lactoglobulin, the major whey protein, could be engineered to bind and protect a wide range
of volatile and unstable flavors during food manufacturing or to release them in more or less
controlled ways by chemical or physical modifications.
Processing significantly affects the interactions between flavors and proteins. Studies on
the interactions between whey protein isolate and flavor compounds (2-nonanone, 1-nonanal,
and trans-2-nonenal). It was found that hydrophobic interactions are weakened upon heating
or high pressure denaturation, whereas covalent interactions were enhanced [30]. Conditions
of high hydrostatic pressure treatment (HHP) and flavor species significantly affect flavor
retention. HHP treatments did not result in significant changes in the flavor retention for
benzaldehyde in WPC solutions, but significant decreases of nonanone retention were only
observed at 100 ppm when WPC solutions were HHP-treated for 10 min. While use of HHP
treatment of WPC has potential in real food systems, these findings demonstrate the
188 Hong Lin, Lisha Wu and Shuhui Wang

importance of careful selection of HHP treatment times and flavor concentrations for desired
outcomes in food application [31].
Milk protein-flavor interactions are very dependent on the conformational state of a
protein. Therefore, factors such as pH, temperature, and high pressure that influence protein
conformation can markedly change flavor binding characteristics of proteins. Interactions of
vanillin with soy, casein, and whey proteins Bindings of vanillin with casein and whey
protein were enthalpy driven, while the interaction of vanillin with soy protein was highly
entropy driven. The researchers inferred that conformational changes of soy protein might be
important in binding of vanillin [32].

REFERENCES
[1] Guang B, Lin H, Wang GC. Food Protein Chemistry. Beijing: Chemical industry press,
2005.
[2] Murray RK, Granner DK, Rodwell VW, Mayes PA. Harper's Biochemistry. 25th
edition. McGraw-Hill Companies, 1999.
[3] http://www.hemaquest.com/indications/hemoglobin.asp.
[4] Xie, BJ. Food chemistry. 1st edition. Beijing: Science Press, 2004: 255.
[5] Damodaran S. Influence of protein conformation on its adaptability under chaotropic
conditions. International Journal of Biological Macromolecules, 1989, 11:2-8.
[6] Han JM, Ledward DA. High pressure/thermal treatment effects on the texture of beef
muscle. Meat Science, 2004, 68: 347-355.
[7] Wang DF. Food Chemistry. 1st edition. Beijing: Chemistry Industry Press. 2007: 118.
[8] Xie BJ. Food chemistry. 1st edition. Beijing: Science Press, 2004: 259.
[9] Xie BJ. Food chemistry. 1st edition. Beijing: Science Press, 2004: 260.
[10] Hu WW, Xie BJ. Food Chemistry. 1st edition. Beijing: Science Press, 1992: 165.
[11] Damodaran S. Interfaces, protein films, and foams. Advances in Food and Nutrition
Research. 1990, 34: 1-79.
[12] Xie BJ. Food chemistry. 1st edition. Beijing: Science Press. 2004: 280.
[13] Wang DF. Food Chemistry. 1st edition. Beijing: Chemistry Industry Press. 2007: 118.
[14] Xie BJ. Food chemistry. 1st edition. Beijing: Science Press. 2004: 278.
[15] Xie BJ. Food chemistry. 1st edition. Beijing: Science Press. 2004: 281.
[16] Pusztai A, Clarke EMW, Grant G, King TP. The toxicity of Phaseolus vulgaris lectins.
Nitrogen balance and immunochemical studies. Journal of the Science of Food and
Agriculture. 1981, 35: 701-711.
[17] Zhu H, Damodaran S. Effects of calcium and magnesium ions on aggregation of whey
protein isolate and its effect on foaming properties. Journal of Agricultural and Food
Chemistry. 1994, 42: 856-862.
[18] Eggum BO, Beames RM. The nutritive value of seed proteins. In: Gottschalk W,
Müller PH. Seed Proteins: Biochemistry, Genetics, Nutritive value. Berlin: Springer.
1983: 499-531.
[19] FAO/WHO/UNU. Energy and Protein Requirements, Report of a Joint
FAO/WHO/UNU Expert Consultation. World Health Organization Technical Report
Series. 1985: 724.
Proteins 189

[20] Lund D. Effect of commercial processing on nutrients. In: Karmas E, Harris RS.
Nutritional Evaluation of Food Processing. New York: Van Nostrand Reihhold. 1988:
319-354.
[21] Wang DF. Food Chemistry. 1st edition. Beijing: Chemistry Industry Press. 2007: 124.
[22] Hurrell RF, Carpenter KJ. Mechanisms of heat damage in proteins. The role of sucrose
in the susceptibility of protein foods to heat damage. British Journal of Nutrition. 1977,
38: 285-297.
[23] Je JY, Park PJ, Kim, SK. Antioxidant activity of a peptide isolated from Alaska pollack
(Theragra chalcogramma) frame protein hydrolysate. Food Research International.
2005, 38: 45-50.
[24] Kim SY, Je JY, Kim SK. Purification and characterization of antioxidant peptide from
hoki (Johnius belengerii) frame protein by gastrointestinal digestion. The Journal of
Nutritional Biochemistry. 2007, 18: 31-38.
[25] Song R, Wei RB, Zhang B, Yang ZS, Wang DF. Antioxidant and Antiproliferative
Activities of Heated Sterilized Pepsin Hydrolysate Derived from Half-Fin Anchovy
(Setipinna taty). Marine Drugs. 2011, 9: 1142-1156.
[26] Song R, Wei RB, Zhang B, Yang ZS, Wang, DF. Optimization of the Antibacterial
Activity of Half-Fin Anchovy (Setipinna taty) Hydrolysates. Food and Bioprocess
Technology. 2011: 1-11.
[27] Rajapakse N, Mendis E, Jung WK, Je JY, Kim SK. Purification of a radical scavenging
peptide from fermented mussel sauce and its antioxidant properties. Food Research
International. 2005, 38: 175-182.
[28] Virtanen T, Pihlanto A, Akkanen S, Korhonen H. Development of antioxidant activity
in milk whey during fermentation with lactic acid bacteria. Journal of Applied
Microbiology. 2007, 102: 106-115.
[29] Minelli A, Bellezza I, Grottelli S, Galli F. Focus on cyclo(His-Pro): history and
perspectives as antioxidant peptide. Amino acids. 2008, 35: 283-289.
[30] Kühn J, Considine T, Singh H. Binding of Flavor Compounds and Whey Protein
Isolate as Affected by Heat and High Pressure Treatments. Journal of Agricultural and
Food Chemistry. 2008, 56: 10218-10224.
[31] Liu XM, Powers JR, Swanson BG, Hill HH, Clark S. High Hydrostatic Pressure
Affects Flavor-binding Properties of Whey Protein Concentrate. Journal of Food
Science. 2005, 70: C581-C585.
[32] Li Z, Grün IU, Fernando, LN. Interaction of Vanillin with Soy and Dairy Proteins in
Aqueous Model Systems: A Thermodynamic Study. Journal of Food Science. 2000,
65: 997-1001.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 6

VITAMINS

Yibin Zhou1, Dongfeng Wang2 and Ping Dong2


1
College of Tea and Food Technology, Anhui Agriculture University, China
2
College of Food Science and Engineering,
Ocean University of China, China

ABSTRACT
Vitamin is one important group of nutrients that determines the nutritional value of
foods. These compounds play extremely important role in many metabolisms in organism
as cofactors of various enzymes and long-term vitamin deficiency can cause various
diseases and even death. Besides, some vitamins, such as Vc and sulfur-containing
compounds, also markedly affect the color and flavor of foods. This chapter introduces
the structures, properties, physiological functions and deficiency symptoms, and stability
of the vitamins that have been identified one by one. Then, the factors that influence
vitamin contents in foods and the changes of vitamins during processing and storage are
discussed.

1. CLASSIFICATION OF VITAMINS
Vitamins are classified by their biological and chemical activities, as well as the
chronological order of being found, but not their structures. Basically each "vitamin" has a
multiple chemical substances (called vitamers) that show similar biological activity, rather
than having a fixed chemical structure. For example, Vitamin A includes retinol, retinal, and
four known carotenoids. The classification and some functions of vitamins are summarized in
Table 6-1.
Vitamins are classified into two water-soluble and fat-soluble ones according to
solubility. Among the 13 vitamins identified, the eight members of the B family and vitamin
C are water soluble, while vitamins A, D, E, and K are fat soluble.
192 Yibin Zhou, Dongfeng Wang and Ping Dong

Table 6-1. Classification of vitamins and their functions

Category Name Synonyms Physiological functions


Water- VB1 Thiamine, Antineuritic factor Antineuritic Prevent beriberi
soluble
VB2 Riboflavin Prevent inflammation of lip and tongue,
Facilitate growth
VB3,VPP Nicotinic acid, Niacin Nicotinamide Prevent pellagra Form Coenzyme I or II
Antipellagra factor
VB6 Pyridoxine, pyridoxal, pridoxal Play a role in amino acid metabolism
VB9 Folic acid, folinic acid Prevent pernicious anemia
VB12 Cyanocobalamin, Prevent pernicious anemia
hydroxycobalamin,
methylcobalamin
VB7,VH Biotin Prevent skin disease Facilitate lipid
metabolism
V B5 Pantothenic acid Prevent Paresthesia
VC Ascorbic acid Prevent or cure scurvy Facilitate the
growth of intercellular substance
Fat-soluble VA1,2 Antixerophthalmic vitamin, Work as a light-sensitive substance in a
carotenoids, visiual cell, prevent epidermal cell
retionol, retinal keratinization, facilitate growth, prevent
and cure xerophthalmia
VD1,3 Cholecalciferol, Lexacalcitol, Regulate metabolism of calcium and
Resistant rickets phosphate, prevent rachitis and
gibbosity
VE Tocopherol, Aantisterility vitamin Prevent barrenness
VK1,2,3 Phytomenadione, phylloquinone Accelerate blood coagulation

Because water-soluble vitamins can dissolve in body fluid and can not be stored readily,
consistent daily intake is required. On the other hand, fat-soluble vitamins, absorbed through
the intestinal tract with the help of lipids (fats), can be easily accumulated in the body. Water-
soluble vitamins are readily excreted from the body so their consumption can be simply
checked by the degree of urinary output.

2. VITAMINS IN FOODS
As described above, vitamins are a class of natural organic compounds that maintains the
normal physiological functions of human body. Although, the amount required for daily diet
is only several micrograms or milligrams, they are essential for human health.
Microorganisms in intestines can synthesize some vitamins, but the quantity far from the need
of human body. Therefore, vitamins have to be largely provided by foods.

2.1. Vitamin A

Vitamin A, including vitamin A1 (retinol) and vitamin A2 (3-dehydroretinol), is also


called antixerophthalmic vitamin, and its deficiency can cause night-blindness,
Vitamins 193

hyperkeratosis, and keratomalacia. Vitamin A is needed by the retina of the eye as the light-
absorbing molecule of retinal, without which the vision will be lost. An oxidized form of
vitamin A – retinoic acid, is a hormone-like growth factor for epithelial and other cells.
Vitamin A was discovered in 1917 by Elmer McCollum (University of Wisconsin–Madison),
Lafayette Mendel (Yale University) and Thomas Burr Osborne (Yale University). In 1919,
Steenbock (University of Wisconsin) proposed that yellow plant pigments (beta-carotene)
might be vitamin A, which was named as ―fat-soluble factor A". Vitamin A exists in animal
tissues, plant tissues and fungi.
(1) Structure.
Structures of vitamin A and its precursors are shown in Table 2.

Table 6-2. Carotenoids structure and activity of vitamin A [1]

Compounds Structure Relative


activity

β-carotene 50

α-carotene 25

β-Apo-8´- 25-30
carotenal

Zeaxanthin 0

Canthaxanthin 0

Astacin 0

Lycopene
0

Both vitamin A1 (retinol) and vitamin A2 (3-dehydroretinol) have the unsaturated


monohydric alcohol (an isoprenoid chain, also called a retinyl group) with the beta-ionine
ring shown as follows:
Both the retinyl group and the beta-ionine ring are essential for the activities of vitamin
A. The difference between vitamin A1 and vitamin A2 lies in the bond between C3 and C4 in
194 Yibin Zhou, Dongfeng Wang and Ping Dong

the beta-ionine ring. Usually vitamin A refers to vitamin A1, retinol. The biological effect of
vitamin A2 is about 40% of that of vitamin A1.
In foods of animal origin, the major form of vitamin A is an ester, primarily retinyl
palmitate, which is converted to retinol in small intestine. Retinol can be converted to its
visually active aldehyde form, retinal. The associated acid (retinoic acid), a metabolite that
can be irreversibly degraded from vitamin A, has only partial vitamin A activity, and does not
function in the retina for the visual cycle.
Food carotenoids, mainly include beta-carotene and lutein (Table 6-2), are also
precursors of vitamin A. Usually they have two connected retinyl groups, which are used in
the body to contribute to vitamin A activity. The well-known orange pigment of carrots (beta-
carotene), which shows some vitamin activities itself, is an important source of vitamin A.
(2) General properties.
Usually, vitamin A is a light yellow viscous liquid. Pure vitamin A1 is a light-yellowish
(or primrose yellow) crystal and melts at 62°C-64°C. Vitamin A is fat soluble and is stable
under alkali conditions or inert atmosphere. Vitamin A is thermal stable and is not destroyed
in temperatures up to 120°C-130°C, but it is liable to decomposition in exposure to air,
oxidants and ultraviolet radiation. Both vitamin A and carotene can react with antimony
trioxide in ethanol to produce dark blue yellow. This property has been used for the
spectrometric determination of vitamin A and carotene.
(3) Physiological functions and deficiency symptoms.
Vitamin A promotes the growth of human body. Besides, it maintains the integrity and
health of epithelial tissue and ensures the normal vision.
Deficiency of vitamin A for a long time can cause night blindness and loss of dark
adaptation. In subjects suffering severe vitamin deficiency, the keratinized epithelial cells at
gastrointestinal tract and eye cornea can even not be produced. The obvious characteristic of
vitamin A deficiency is xerophthalmia, such as cornea hyperemia, inflammation and sclerosis.
This is why vitamin A is also known as the antixerophthalmic vitamin. Excessive intake of
vitamin A (more than 65,000~500,000 international units) can result in poisoning, but the
symptoms can disappear several days later if vitamin A is excluded from the diet.
(4) Stability and degradation.
All naturally occurring carotenoids are present in the trans conformation. The compounds
can transform to the cis conformation and lose its activity when heating. The isomerization of
carotenoids occurs readily during storage, as shown in Table 6-3. In addition, exposure to
light, acids and watery iodine also causes the transform from cis- to trans-conformation of
carotenoids and retinols.
The destruction mechanisms of vitamin A precursors vary with processing conditions
(Figure 6-1). The cis-trans transformation of beta-carotenes upon heating occurs only in the
absence of oxygen. Beta-carotene is decomposed to a series of aromatic hydrocarbons at
higher temperatures and the major decomposition product is lonene, which has important
implications on food flavor. Carotene is often intentionally added to foods to produce aroma
during food processing.
Vitamins 195

Table 6-3. Distribution of β-carotene isomers in fresh fruits and


processed vegetables [1]

Products Status Percentage of total β-carotene (%)


13-cis All trans 9-cis
Sweet potato Fresh 0 100.0 0
Canned 15.7 75.4 8.9
Carrot Fresh 0 100.0 0
Canned 19.1 72.8 8.1
Pumpkin Fresh 15.3 75.0 9.7
Canned 22.0 66.6 11.4
Spinage Fresh 8.8 80.4 10.8
Canned 15.3 58.4 26.3
Collard Fresh 16.3 71.8 11.7
Canned 26.6 46.0 27.4
Cucumber Fresh 10.5 74.9 14.5
Pickld cucumber Pasteurized 7.3 72.9 19.8
Tomato Fresh 0 100.0 0
Canned 38.8 53.0 8.2
Peach Fresh 9.4 83.7 6.9
Canned 6.8 79.9 13.3
Apricot Dehydrated 9.9 75.9 14.2
Canned 17.7 65.1 17.2
Nectarine Fresh 13.5 76.6 10.0
Plum Fresh 15.4 76.7 8.0

Figure 6-1. β-Carotene degradation [2].


196 Yibin Zhou, Dongfeng Wang and Ping Dong

Table 6-4. Contents of beta-carotene in fresh and dehydrated carrot [3]

Carrot Content (mg/kg)


Fresh 980-1860
Vacuum freeze-drying 870-1125
Conventional air-drying 636-987

Oxygen, light, enzymes, and lipid hydroperoxide enhance the oxidative degradation of
carotene. Oxidation of beta-carotene first involves the formation of the 5, 6-epoxide, which
will isomerize to beta-carotene oxide, namely 5, 8-epoxide. Vitamin A and provitamin A are
usually inactivated in dehydrated foods, but the loss varies with the drying technology (Table
6-4). Maintenance of Aw and oxygen partial pressure in low levels can reduce the loss of
carotenoid and vitamin A.

2.2. Vitamin D

Vitamin D is also known as anti-rickets or anti-rachitis vitamin. Six members have been
ascertained, including Vitamin D2, Vitamin D3, Vitamin D4, Vitamin D5, Vitamin D6, and
Vitamin D7, among which Vitamin D2 and Vitamin D3 are the most important.
Vitamin D in food always coexists with vitamin A. Fat in fishes or liver in animals are
rich in vitamin D. The content of Vitamin D in fish oil from seafood is much high, followed
by egg yolks, milk, and butter. Sunlight can promot the synthesis of vitamin D in animals.
Thus, the levels of vitamin D in milk and cream produced in summer are more than those in
winter due to the stronger sunlight in summer.
(1) Structure.
The vitamin D members are steroids that contain the cyclopentanoperhy drophenanthrene
ring structure and they are distinguished by their structures of the branch chain R, as shown in
Figure 6-2. Vitamin D is found only in animal tissues, but most plants contain sterols, which
can be converted to vitamin D by ultraviolet light.
(2) General properties.
Vitamin D members are colorless crystal and fat soluble. Vitamin D is quite stable
against acids, alkali, and oxygen, but is readily destroyed by light and temperatures higher
than 160°C.
(3) Physiological functions and deficiency.
Vitamin D regulates the metabolism of calcium and phosphorus and maintains the level
of calcium and phosphorus in blood. Hence, the deficiency of calcium and phosphorus is
often related to the level of vitamin in human body. The increased requirements of pregnant
and lactating women on calcium and phosphorus can be realized by vitamin D-reinforced
diets.
Vitamin D deficiency leads to osteomalacia and rickets. However, excessive intake of
vitamin D can cause posioning. The symptoms of acute vitamin D poisoning include loss of
appetite, nausea, vomiting, diarrhea, headache and polyuria and those of chronic poisoning
include weight loss, albinism, and even soft-tissue calcification or serious kidney failure.
Vitamins 197

R
CH 3
19
11 12 13 17 16
CH 2 15
9 14
8
1 10
2
3 5 7
4 6
HO

CH2
Vitamin D2 R= CH CH CH CH CH
CH3 CH3 CH3
CH2
R= CH CH2 CH2 CH2 CH
Vitamin D3
CH3 CH3

CH3
R= CH CH2 CH2 CH CH
Vitamin D4
CH3 CH3 CH3

CH3
Vitamin D5 R= CH CH2 CH2 CH CH
CH3
CH3 CH 2C H 3

CH3
R= CH CH CH CH CH
Vitamin D6
CH3 CH3
C H 2C H 3

CH3
Vitamin D7 R= CH CH2 CH CH2 CH
CH3 CH3
CH3

Figure 6-2. Chemical structures of vitamin D members [4].

2.3. Vitamin E

Vitamin E is also called as anti-sterility factor or tocopherol and is a family of eight


separate compounds, in which, α-, β-, γ-, δ-tocopherols are the most important and α-
tocopherol is the most active. Hence, tocopherol sometimes refers solely to α-tocopherol.
The contents of tocopherol in some foods are listed in Table 6-5. The content of vitamin
E is often expressed as international unit (IU). One IU of vitamin E is equal to 1 mg of D, L-
α-tocopherol acetate or 0.667 mg of D-α-tocopherol.
(1) Structure.
Vitamin E is derivatives of benzo-3 H-pyran and its general structure is shown in Figure
6-3. The members of the vitamin E family differ in the structures of the three side chains, as
listed in Table 6-6.
(2) General properties.
Pure vitamin E is a transparent and light yellow oily liquid. It is fat soluble and is stable
against acid, alkaline and heating (up to 200°C in the absence of oxygen), but it is susceptible
to oxidation by oxygen. Hence, vitamin E is an important natural antioxidant in foods.
Vitamin E is quiet stable under white light, but is easily destroyed by ultraviolet light.
(3) Physiological function and deficiency.
198 Yibin Zhou, Dongfeng Wang and Ping Dong

Table 6-5. Contents of tocopherol in plant oil and some foods [4,5]

Foods α-T α-T3 β-T β-T3 γ-T γ-T3 δ- T3


Plant oil (mg/100g)
Sunflower seed 56.4 0.013 2.45 0.207 0.43 0.023 0.087
Peanut oil 0.013 0.007 0.039 0.394 13.1 0.03 0.922
Bean oil 17.9 0.021 2.80 0.437 60.4 0.078 37.1
Cottonseed oil 40.3 0.002 0.196 0.87 38.3 0.089 0.457
Corn germ oil 27.2 5.37 0.214 1.1 56.6 6.17 2.52
Olive oil 9.0 0.008 0.16 0.417 0.471 0.026 0.043
Palm oil 9.1 5.19 0.153 0.4 0.84 13.2 0.002
Other foods (μg/ml or g)
Spinage 26.05 9.14
Beef 2.24
Flour 8.2 1.7 4.0 16.4
Barley 0.02 7.0 6.9 2.8
Notes: T-tocopherol; T3- tocotrienol.

Figure 6-3. Structure of vitamin E [6].

Table 6-6. Members of the vitamin E family and their


Physiological values [4,7]

Relative
Side chain Name R1 R2 R3 Occurrence
biological value
α- tocopherol -CH3 -CH3 -CH3 Wheat embryo 1
β- tocopherol -CH3 -H -CH3 Wheat embryo 0.5
γ- tocopherol -H -CH3 -CH3 Corn 0.2
[ ]3
δ- tocopherol -H -H -CH3 Bean 0.1
Σ- tocopherol -CH3 -CH3 -H Rice 0.5
η- tocopherol -H -CH3 -H Rice 0
ε- tocopherol -CH3 -H -CH3 Corn 0.5
[ ]3
Σ1- tocopherol -CH3 -CH3 -CH3 Rice 0.5

Vitamin E is involved in the conversion of arachidonic acid to prostaglandins and slows


down the aggregation of blood platelets. Vitamin E deficiency can cause damage of the
genitals of human and infertility.
Vitamins 199

Figure 6-4. Oxidation pathway of α-tocopherol initiated by peroxide free radicals [6].

(4) Stability and degradation.


The majority of vitamin E in foods is lost during processing and storage. For example,
about 80% vitamin E is lost in the processing of wheat, corn, oat and rice. Degreasing,
dehydration, lipid refinement and oxidation significantly affect vitamin E contents in foods,
but the specific losses depend on the processing technology and starting material. For
example, 36~45% of α-tocopherol is lost in the dehydration of chicken and beef, but there is
almost no loss in the process of dehydration of pork. Rinsing with water does not reduce
tocopherol contents in foods due to it fat solubility. The stability of vitamin E in foods is
related to aw, temperature and duration during storage.
Vitamin E is a strong antioxidant and can retard or prevent lipid oxidation. In the
presence of peroxide free radicals, vitamin E is oxidized to α-tocopherol oxides, α-tocopherol
quinone and α-tocopherol hydroquinone, as shown in Figure 6-4.
In addition, singlet oxygen can also attack the epoxy system in tocopherol molecule,
resulting in the formation of a hydrogen peroxide derivative. The derivative can generate
tocopherol quinone and α-tocopherol quinone-2-3-epoxide after rearrangement (Figure 6-5).

2.4. Vitamin K

Vitamin K is found due to its blood coagulation functions. Two kinds of vitamin K have
been identified, including vitamin K1, which is extracted from alfalfa, and vitamin K1, which
is obtained from degraded fishes.
200 Yibin Zhou, Dongfeng Wang and Ping Dong

Figure 6-5. Oxidation pathway of α-tocopherol initiated by singlet oxygen [7].

Figure 6-6. Structures of vitamin K1 and vitamin K2.

Vitamin K occurs mainly in plant tissues. Green leafy vegetables, such as alfalfa,
cabbage, cauliflower, spinach and cabbage, contain more vitamin K1 than vitamin K2, with
the latter derived mainly from the metabolism of intestinal bacteria.
(1) Structure.
Vitamin K1 and K2 are the derivatives of 2-methyl-1, 4-naphthoquinone and the two
compounds differ in their side chains, as shown in Figure 6-6.
(2) General properties.
Vitamin K1 is a viscous yellow oily liquid can be crystallized from alcohol solution
under low temperature, and its melting point is -20 °C. Vitamin K2 is a yellow crystal and
melts in the temperature range from 53.5°C to 54.5°C. Both vitamin K1 and K2 are stable
Vitamins 201

against heating, but are readily destroyed by alkali and sunlight. K2 is more susceptible to
oxidation than K1.
(3) Physiological function and deficiency.
Vitamin K promotes blood coagulation and is essential for the synthesis of prothrombin
in liver. Vitamin K deficiency leads to reduced content of prothrombin in plasma and
prolonged duration of blood coagulation. Vitamin K will lose its promotion of prothrombin
synthesis in liver if liver dysfunction occurs. In addition, vitamin K also enhances intestines
peristalsis and secretion.

2.5. Vitamin B1

Vitamin B1 is also known as thiamin. Yeast is reported to have the highest content of
vitamin B1, followed by muscle, drupe, and eggs. Vitamin B1 in cereals occurs mainly in
cortex and embryo. Table 6-7 lists the contents of vitamin B1 in some cereal products.
(1) Structure.
Vitamin B1 contains a pyrimidine ring that contains an amino group and a thiazole ring
thatn contains a sulfur residue, as shown in Figure 6-7.
(2) Properties.
The hydrochloride salt of vitamin B1 is a white crystal. The crystal is stable at 100°C for
24 hours, but is decomposed in temperatures higher than 249°C. Vitamin B1 is water soluble
and one gram of vitamin B1 can dissolve in 1mL water, 18 mL glycerin, 100 mL 95% alcohol
or 315 mL anhydrous alcohol, but is insoluble in ethyl ether, acetone, chloroform or benzene.
Vitamin B1 has a smell similar to that of yeast and tastes bitter.

Table 6-7. Contents of vitamin B1 in some cereal products


(mg/100g Dry)[8]

Name Vitamin B1 content Names Vitamin B1 content


Wheat 0.37-0.61 Brown rice 0.3-0.45

Bran 0.7-2.8 Cortex 1.5-3.0


Embryo 1.56-3.0 Rice embryo 3.0-8.0
Flour (85% yield) 0.3-0.4 Endosperm 0.03
(73% yield) 0.07-0.1 Corn 0.3-0.45
(60% yield) 0.07-0.08 Soybean 0.1-0.6
Potato 0.08-0.1 Pea 0.36

Figure. 6-7. Structure of thiamin.


202 Yibin Zhou, Dongfeng Wang and Ping Dong

Figure 6-8. Structure of thiochrome.

Figure 6-9. Structure of thiamin pyrophosphate (TPP).

Vitamin B1 is stable in solutions with pH less than 3.5. Under these conditions, the
vitamin can maintain its activity even when heated at 120°C; however, it is easily destroyed
when the pH exceeds 3.7. For example, 25% of thiamin is lost when heated in 97°C at pH 4.3
for 1 hour and the value reaches 80% when the pH is further increased to 7.0. Though vitamin
B1 is readily damaged by high temperatures in alkaline conditions, cooking is insufficient to
completely destroy the vitamin in rice. During bread bakery, about 15~30% of the total
amount of vitamin B1 in flour is lost.
Both vitamin B1 and its pyrophosphate salt can be oxidized by mild oxidant (such as the
alkaline solution of potassium ferricyanide) to produce the dark blue fluorescence-emitting
thiochrome. This reaction has been used to quantitative determination of vitamin B1. The
structure of thiochrome is shown in Figure 6-8:
Thiamine can be converted to thiamine pyrophosphate (TPP, Figure 6-9) by thiamine
kinase in the presence of ATP. TPP plays an important role in sugar metabolism.
(3) Physiological function and deficiency.
Vitamin B1 participates in the oxidative decarboxylation of α-keto acid, such as pyruvic
acid and α- ketoglutaric acid, as cocarboxylase. Vitamin B1 deficiency results in the
accumulation of pyruvic acid in blood. Meanwhile, the accumulated pyruvic acid inhibits the
activity of dehydrogenase and leads to the accumulation of lactic acid. Hence, vitamin B1 is
essential for sugar metabolism.
Vitamin B1 is of special importance for consumers with starch as the main energy source.
It is reported that the amount of vitamin B1 required is propontional to the amount of
carbohydrates consumed and it is recommended that 1 microgram of vitamin B1 is needed for
the metabolism of 1 gram of carbohydrates.
The early symptom of vitamin B1 deficiency includes nervous system disorder, mental
and physical fatigue, dyspepsia and loss of appetite. If vitamin B1 cannot be supplemented in
time, the patients develop polyneuritis or beriberi. In this stage, the patients become poor in
heath status and other symptoms include lower limb edema, nerve paralysis, loss of muscle
contraction capacity, and even death.
(4) Stability and degradation.
Vitamin B1 is the most unstable among all the vitamins identified and is susceptible to the
influence of aw, pH, temperature, ionic strength, buffer solution and the presence of other
compounds. The degradation of vitamin B1 is initiated by the nucleophilic substitution of the
Vitamins 203

methylene carbon between the two rings. Therefore, vitamin B1 can be readily destroyed by
strong nucleophiles such as HSO3-.
Vitamin B1 is stable at low aw and room temperature. For example, nearly no loss of
vitamin B1 is observed when cereal foods with aw ranging from 0.1 to 0.65 are stored at
37°C. In storage temperature 45°C and aw 0.2~0.5, the decomposition of vitamin B1
increases. The maximum vitamin B1 degradation is observed in aw 0.5. As the aw further
increases, the loss of vitamin B1 decreases, as shown in Figure 6-10.
Temperature significantly affects the retention of vitamin B1 during storage (Table 6-8).
In 1.5°C, no loss of vitamin B1 is detected in tomato juice, pea and orange juice after storage
for 12 months. When the storage temperature is elevated to 35°C, up to 50% of vitamin B1 is
lost.
The relationship between vitamin B1 degradation rate and pH is shown in Figure 6-11.
The same relationship is observed in either phosphate buffer or cereal. That is, the
degradation of vitamin B1 increases with the rise of pH.
Similar to other other-water soluble vitamins, rinsing and boiling cause great loss of
vitamin B1. Table 6-9 lists the effect of various processing methods on the retention of
vitamin B1 in various foods.
The thermal degradation of Vitamin B1 can generate special flavor compounds, such as
hydrogen sulfide, furan, and dihydro-thiophenol. These compounds impart foods with special
flavor.

2.6. Vitamin B2

Vitamin B2 is also known as riboflavin and is found mainly in yeast, liver, milk, muscle
and yolk. In addition, Greenery vegetables, grain and germinated seed also contain vitamin
B2.

Figure 6-10. Relationship between vitamin B1 degradation rate and aw of cereal foods when stored at
45°C [9].
204 Yibin Zhou, Dongfeng Wang and Ping Dong

Table 6-8. Retention of vitamin B1 in some foods during storage [10]

Species Retention after storage Species Retention after storage for 12


for 12 months (%) months (%)
1.5°C 35°C 1.5°C 35°C
Apricot 35 72 Tomato juice 60 100
Mungbean 8 76 Pea 68 100
Lima bean 48 92 Orange juice 78 100

Table 6-9. Effect of treatment methods on the retention of vitamin B1 [11]

Products Treatment Retention (%)


Cereal Extrusion, Cook 48-90
Potato Fried after immerse in water for 16 hr 55-60
Fried after immerse in sulfurous acid solution for 16 hr 19-24
Soybean Boiled in water or carbonate after immerse in water 23-52
Smashed potato Various heat treatments 82-97
Vegetable Various heat treatments 80-95
Freeze or fried Various heat treatments 77-100
fish

Figure 6-11. Relationship between vitamin B1 degradation and pH.


Vitamins 205

(1) Structure.
Riboflavin is the condensation product of D-ribitol and 7, 8-dimethyl-isoalloxazine. The
structure of vitamin B2 is showed in Figure 6-12.
(2) General properties.
Riboflavin is an orange needle crystal and tastes bitter taste. This vitamin melts in 282 °C
and sparingly soluble in water, but readily soluble in alkaline solutions.
The aqueous solution of riboflavin generates yellow fluorescence in wavelength range
440~500nm. This property can be used to spectrometrically determine the content of the
compound.
Riboflavin is more thermal stable than vitamin B1. After dried yeast is heated in 120 °C
for several hours, nearly all vitamin B1 is lost, but vitamin B2 is completely retained.
Riboflavin is unstable in the presence of light and ultraviolet, especially in alkaline and
high temperature conditions. In the presence of light, riboflavin in alkaline solutions is
converted to lumiflavin. This product has the same color and fluorescence emission capability
as riboflavin. In acidic or neutral solutions, riboflavin is transformed to lumichrome, which
can emit blue fluorescence. The structures of the two compounds are illustrated in Figure 6-
13.
(3) Physiological function and deficiency.
Vitamin B2 is an essential cofactor for enzymes involved in cellular respiration and
dehydrogenation. Vitamin B2 deficiency leads to reduced respiration capacity and retarded
metabolism. The symptoms of vitamin B2 deficiency include stomatitis, angular cheilitis, eye
keratitis, and dermatitis.
(4) Stability and degradation.
Riboflavin is stable against thermal treatment and oxygen. Riboflavin is resistant to light
in acidic solutions, but is readily decomposed by light in alkaline solutions. In light-induced
decomposition, photochemical degradation occurs in ribitol to generate inactive lumiflavin
and a series of free radicals (Figure 6-14). Lumiflavin has stronger oxidizability than
riboflavin and its generation accelerates the destruction of other vitamins, especially Vc. The
special off-odor of sunlight-exposed milk has been attributed to the light-induced
decomposition of riboflavin.

Figure 6-12. Structure of vitamin B2.


206 Yibin Zhou, Dongfeng Wang and Ping Dong

Figure 6-13. Chemical structures of lumiflavin and lumichrome [12].

Figure 6-14. Photodegradation of riboflavin [12].

2.7. Pantothenic Acid

Pantothenic acid (Vitamin B5) is a water soluble vitamin and occurs widely in foods,
including yeast, liver, kidney, egg, muscle, skimmed milk, pea, peanut and sweet potato. The
contents of pantothenic acid in unpolished rice, wheat, and corn are about 1.7 mg/100g, 1.0-
1.5 mg/100g, and 0.46 mg/100g respectively. Intestinal bacteria and some plants can
synthesize pantothenic acid.
(1) Structure.
The chemical name of pantothenic acid is N-(α, γ-dihydroxy-β, β-dimethyl-butyryl)- β-
alanine. Its structure is shown in Figure 6-15.
(2) Properties.

CH3
HOCH2 C CH CO NH CH2 CH2 COOH
CH3 OH

Figure 6-15. Structure of vitamin B5.


Vitamins 207

4 COOH CONH2
5 3
61 2
N N
Nicotinic acid Nicotinamide

Figure 6-16. Structures of nicotinic acid and nicotinamide.

Pantothenic acid is light yellow and viscous. It is soluble in acetic acid and water, but
insoluble in chloroform and benzene. In neutral solutions, pantothenic acid is resistant to heat,
oxidation and reduction, but can be decomposed β-alanine and other compounds by acids,
bases, and dry heating. The calcium salt of pantothenic acid is a colorless crystalline powder.
Calcium pantothenate tastes slightly bitter and is water soluble. It is stable against light and
air, but its aqueous solutions of pH 5~7 can be destroyed by heat.
In organisms, pantothenic acid takes effect mainly in the form coenzyme A (CoA) form.
(3) Physiological function.
Pantothenic acid is the coenzyme of many enzymes involved in carbohydrate, lipid and
protein metabolism and plays an important role in the transfer of the acetyl group.
Pantothenic is also essential for the growth of some microorganisms.
(4) Stability and degradation.
Pantothenic acid is stable in neutral solutions, but is easily decomposed in acid solutions.
In the pH range 6~4, its decomposition rate increases with the increase of acidity. Due to the
heat liability and strong hygroscopicity of pantothenic acid, the calcium salt of pantothenic
acid has been commercially used as an alternative. The changes of pantothenic acid contents
in foods are affected by processing methods in addition to the materials. For example, the loss
of pantothenic acid in pasteurized milk is usually less than 10%. The loss of pantothenic acid
in vegetables is about 10-30 % and most of vitamin is lost during rinsing. The bioavailability
of dietary pantothenic acid is about 51%.

2.8. Vitamin B3

Vitamin B3 is known as vitamin PP and is famous for its anti-pellagra capability. Two
members of the family have been identified, including nicotinic acid and nicotinamide. Both
te two compounds occur widely in yeasts, liver, muscle, milk, peanut, and soybean. The
cortex and embryo of cereal grains also contain high levels of the two compounds.
(1) Structure.
Both nicotinic acid and nicotinamide are the derivates of pyridine and their structures are
shown in Figure 6-16.
(2) Properties.
Both nicotinic acid and nicotinamide are colorless needle crystals. Nicotinic acid melts in
235.5-236°C. It is sparingly soluble in water but readily soluble in ethanol. The melting point
of nicotinamide is 129-131°C and the compound is readily water soluble. Nicotinic acid is the
most stable among all the vitamins identified and is not destroyed by light, air, heating, acids,
or bases. Nicotinamide is converted to nicotinic acid in acidic solutions upon heating.
208 Yibin Zhou, Dongfeng Wang and Ping Dong

Figure 6-17. Oxidized and reduced states of nicotinamide.

Nicotinic acid can react with cyanogen bromide to yield a yellow-green compound. This
reaction has been used for the quantitative analysis of nicotinic acid and nicotinamide.
The double bond between C4 and C5 can be reduced. Hence, the two compounds can be
present either in oxidized or reduced form.
Nicotinamide implements its in vivo function as nicotinamide dinucleotide (NAD, also
known as coenzyme I) or nicotinamide dinuclotide phosphate (NADP, CoII). NADP can be
converted from NAD by phosphorylation with ATP.
(3) Physiological functions and deficiency.
Nicotinamide is a main component of CoI and CoII, which are the coenzymes of many
dehydrogenases. Nicotinamide in the two enzymes occur either in the reduced and oxidized
states and the transformation between the two states realize the transfer of hydrogen (Figure
17).
Nicotinamide deficiency results in pellagra. The symptoms in the early phase include
symmetrical dermatitis in hands, cheeks, foreheads and other naked areas, gastrointestinal
disorders, mouth and tongue inflammation, indigestion and diarrhea. If nicotinamide is not
supplemented in time, the patients might suffer neurological disorder and even death.
(4) Stability and degradation.
As mentioned previously, nicotinic acid is the most stable vitamin. However, non-
chemical processing of vegetables, such as dressing and rinsing, can cause loss of this vitamin
as other water-soluble ones. The loss of nicotinic acid of pork and beef during storage is
attributed to biochemical reactions, roasting seems to have no effect the content of nicotinic
acid. No loss of nicotinic acid is detected in processed diary products.

2.9. Vitamin B6

Vitamin B6 is also called adermine and includes pyridoxine, pyridoxal, and


pyridoxamine. Vitamin B6 widely distributes in animals and plants. Wheat germ, rice bran,
soybean, peanut, yeast, liver, fish and meat are the major storage pool of this vitamin.
(1) Structure
Pyridoxine, pyridoxal and pyridoxamine are the derivatives of pyridine and their
structures are shown in Figure 6-18.
(2) Properties.
Pyridoxine, pyridoxal and pyridoxamine are colorless crystals and are readily soluble in
water and alcohol. The three compounds are stable in acidic conditions, but are easily
destroyed in alkaline conditions. Pyridoxine is heat tolerant, but the other two compounds are
not.
Vitamins 209

Figure 6-18. Structures of pyridoxine, pyridoxal and pyridoxamine.

Figure 6-19. Structures of pyridoxal phosphate and pyridoxamine phosphate.

Figure 6-20. Structures of Schiff bases of pyridoxal and pyridoxamine [13].

Pyridoxine can be converted to pyridoxal or pyridoxamine and all the three compounds
can be phosphorylated, of which, pyridoxal phosphate and pyridoxamine phosphate (Figure
6-19) are the major active forms of vitamin B6 and they are the coenzymes of amino acid
decarboxylases and amino transferases.
(3) Physiological functions and deficiency.
Vitamin B6 is the coenzyme of amino acid decarboxylases and amino transferases and
participates in many metabolism reactions. Long-term deficiency of vitamin B6 may cause
skin irritation and damage of the central nervous system and hematopoietic organ.
(4) Stability and degradation.
The three members of vitamin B6 exhibit excellent thermal stability, of which,
pridoxamine is the most stable and is selected in food fortification. In the presence of oxygen,
vitamin B6 can be transformed to inactive 4-pyridoxine acid by UV irradiation. Vitamin B6 is
easily degraded in alkaline solutions, but is stable in acidic conditions.
Vitamin B6 can react with amino acids to produce Schiff base when heated (Figure 6-20).
In acidic solutions, the formed Schiff base undergoes further dissociation. In addition, the
Schiff base can rearrange to yield many cyclic compounds.
Vitamin B6 suffers more or less loss during processing. For example, about a half of the
vitamin is lost in sterilized liquid milk and the loss further increases during storage for 7~10
days. High temperature-short time pasteurization (92°C, 2~3s) causes only a loss of 30% in
milk, but the values reaches up to 84% when sterilized in 119~120°C for 13 to 15 min.
210 Yibin Zhou, Dongfeng Wang and Ping Dong

2.10. Vitamin H

Vitamin H is also called biotin and includes α- and β-biotin, of which, the former exists
in yolk and the later in liver. Biotin widely distributes in plant and animal tissues. Most of
vitamin H is bound with proteins and only a little portion occurs in the unbound state. Many
organisms can synthesize biotin on their own. In human body, part of this vitamin is
synthesized by intestinal bacteria.
(1) Structure.
Both the two members of biotin are thionic compounds and they are differentiated by
their side chains, as shown in Figure 6-21.
(2) Properties.
Biotin is a colorless needle crystal and its melting point is 232~233°C. Biotin is soluble
in hot water and ethanol, but is insoluble in ether or chloroform. It is stable against light,
heating, acids, but can be destroyed by high temperatures and oxidants.
(3) Physiological functions and deficiency.
Biotin is the cofactor of many carboxylases. It is the carrier of CO2 in many CO2 fixation
reactions. Because biotin can be synthesized intestinal microorganisms in addition to intake
from foods, biotin deficiency occurs rather occasionally compared with other vitamins. The
symptoms of biotin deficiency include dermatitis, muscle pain, hypersensitivity, fatigue,
anorexia, and slight anemia.

2.11. Vitamin B11

Vitamin B11 is also called as folic acid or folate. Folic acid occurs widely in nature.
Green leaves, liver, kidney, cauliflower, yeast, beef and kernels are found to have high
contents.
(1) Structure.
Vitamin B11 is composed of pteridine, p-aminobenzoic acid and L-glutamate, as shown
in Figure 6-22.
(2) Properties.

Figure 6-21. Structure of biotin [14].


Vitamins 211

Figure 6-22. Structure of vitamin B11.

Folic acid is a bright yellow crystal and is sparingly soluble in water. It is easily
destroyed by light in aqueous solutions. Caron atoms in positions 5, 6, 7, 8 can be reduced by
NADP·H2, positions to yield tetrahydrogen folic acid (THFA).
THFA is the carrier of many one-carbon groups, such as formyl, formaldehyde, and
methyl and plays important role in the synthesis of proteins and nucleic acids.
(3) Physiological function and deficiency.
Folic acid is involved in the synthesis of proteins and nucleic acids. Its deficiency can
cause blood diseases and the symptoms include pernicious anemia, glossitis and
gastrointestinal problems. Because intestinal bacteria can synthesize folic acid, the incidence
of folic acid deficiency is very low.
(4) Stability and degradation.
Dihydrofolate (FH2) and tetrahydrofolate (FH4) are easily oxidized in air and are
sensitive to pH. The two compounds are the most stable in pH ranges 8-12 and 1-2. In neutral
solution, FH4 and FH2, and folic acid are quickly oxidized to p-aminobenzoyl-L-glutamic
acid (PABG), pteridine, xanthine, 6-methyl pterin and many other pterin derivatives. FH4 is
oxidized more quickly in acidic solutions than in alkaline solutions and its oxidation products
include PABG and 7,8-dihydro-pterin-6-carboxyl aldehyde. Reducing agents, such as thiol
and ascorbic acid, can slow down the oxidation of FH2 and FH4.
The stability of several derivatives of THFA follows the decreasing order of 5-formyl-
tetrahydrofolate > 5-methyl-tetrahydrofolate > 10-methyl-tetrahydrofolate > tetrahydrofolate.

Table 6-11. Loss of folic acid in some foods during processing [11]

Foods Processing methods Loss of folic acid activity (%)


Eggs Oil fried or cooking 18~24
Liver Cooking None
Atlantic plaice Cooking 46
Cauliflower Boiling 69
Carrot Boiling 79
Meat γ-radiation None
Grapefruit juice Canned or storage Negligible
Tomato juice Canned 50
Storage in dark for 1 year 7
Exposure to light for 1 year 30
Corn Refining 66
Flour Milling 20~80
Meat or Canned storage for 3 years Negligible
vegetables Canned storage for 5 years Negligible
212 Yibin Zhou, Dongfeng Wang and Ping Dong

Figure 6-23. Structure of vitamin B12..

The loss of folic acid in foods varies with food species and processing technologies, as
listed in Table 6-10.

2.12. Vitamin B12

Vitamin B12 contains cobalt and is therefore also called cobalamine. To present, at least
five members of this family have been identified, but vitamins B12 sometimes refer solely to
cyanoccbalamin.
Liver has the highest vitamin B12 content, followed by milk, meat, egg, and fish.
Vitamin B12 is not found in plants. Because only bacteria can synthesize vitamin B12,
vitamin B12 in animal tissues derives either from foods or intestinal microbes. Vitamin B12
are bound to proteins in foods and are absorbed only after it is dissociated from the complex
by heating or protease hydrolysis.
(1) Structure.
Vitamins of the B12 family are polycyclic compounds and contain one trivalent cobalt
ion (Figure 6-23). According to the structure the R group, cobalamide is divided into
cyanocobalamin (R = CN), hydroxocobalamin (R = OH), adenosylcobalamin (R = Ado),
nitrosocobalamin (R = nitroso), methylcobalamin (R = CH3), etc.
(2) Properties.
The compounds of the B12 family are pink needle crystals and melt in temperatures higer
than 320°C. The compounds are soluble in water, ethanol and propanol, but insoluble in
chloroform. Vitamin B12 in either crystals or aqueous solutions (pH 4.5~5) are quite stable,
but are readily destroyed strong acids/bases, sunlight, oxidants and reducing agents.
(3) Physiological functions and deficiency.
Vitamins 213

Vitamin B12 is the coenzyme of many enzymes and participates in a variety of reactions.
For example, it is the carrier of methyl and is involved in the biosynthesis of methionine and
thymine. Besides, vitamin B12 increases the bioavailability of folic acid.
Vitamin B12 synthesized by intestinal microbes must combine with the special
mucoprotein (also known as endogenous factor) secreted by gastricism for its adsorption.
Lack of the endogenous factor retard the absorption of vitamin B12 and causes pernicious
anemia and lesions in nervous system, tongue and gastricism.
(4) Stability and degradation.
Vitamin B12 in aqueous solutions is stable in pH range 4~6 in dark environments and
room temperature, but it is quantitatively destroyed in alkaline solutions upon heating.
Reducing agents in low levels protect vitamin B12 from destruction, but those in high
concentrations have opposite effects. Trivalent iron salts can stabilize vitamin B12, but
bivalent iron salts rapidly destroy the vitamin.

2.13. Lipoic Acid

Lipoic acid occurs either in the reduced and oxidized forms and their structures are
shown in Figure 6-24). Lipoic acid, which is necessary for the growth of microorganisms and
protozoa and dissolves in water, is generally classified as a vitamin B. Lipoic acid is rich in
liver and yeast, and the human body can synthesize it. Lipoic acid is a coenzyme of pyruvate
dehydrogenase and α-ketoglutarate dehydrogenase, plays the role as acyl transfer.
Lipoic acid is water soluble. It is essential for the growth of microorganisms and protozoa
and is generally considered a member of the B family. Human can synthesize lipoic acid on
their own. Lipoic acid is the coenzyme of pyruvate dehydrogenase and α-ketoglutarate
dehydrogenase and participates in acyl transfer.

2.14. Vitamin C

(1) Structure.
Vitamin C, or ascorbic acid, is an acid derivative of hexose. Vc occurs widely in fresh
fruits and vegetables. Only L-ascorbic acid is bioactive. The structure of vitamin C is shown
in Figure 6-25.
(2) Properties.
Vc is a colorless sliced crystal and melts in 190-192°C. It is soluble in water and ethanol
and its aqueous solutions taste sour. Vc is unstable against heating, sunlight exposure, and
metal ions such as such as Cu2+, Fe3+. However, it is quite stable in oxalic acid or
metaphosphoric acid solutions. Vc is a strong reducing agent and can be reversibly oxidized
to the dehydrogenated form (Figure 6-25). Vitamin C has been widely used as an antioxidant
in the food industry.
L-ascorbic acid can reduce the blue dye 2, 6-dichlorophenol-indophenol to the colorless
leuco and react with 2,4-dinitrophenylhydrazine to yield colored hydrazone. These reactions
can be used for the qualitative and quantitative analysis of Vc.
214 Yibin Zhou, Dongfeng Wang and Ping Dong

Figure 6-24. Structure of lipoic acid [15].

Figure 6-25. Structure of Vitamin C.

(3) Physiological functions and deficiency.


Vc enhances the formation of supportive tissues and intercellular adhesion molecules. Vc
functions as both hydrogen donor and receptor and plays important role in the redox reactions
in human body. In addition, Vc is also involved in disease resistance and detoxification.
Because human body cannot synthesis Vc, Vc deficiency causes a variety of symptoms,
of which, the most notably symptom is scurvy. The symptoms in the early stage of Vc
deficiency include local inflammation of skin, loss of appetite, dyspnea and general fatigue.
In the later phase, the patients might suffer microvessel fracture in visceral, subcutaneous
tissue, metaphyseal and gingiva and even death.
(4) Stability and degradation.
The stability of Vc depends largely on temperature, pH, oxygen, enzymes, metal ions
such as Cu2+ and Fe3+, aw, initial concentration, and Vc to dehydrogenated Vc ratio.
Although the nonenzymatic browning reaction of Vc occurs slowly in anaerobic conditions,
Vc is degraded in the pathway shown in Figure 6-26 in weak acid and weak alkaline
conditions.
Diketo-gulonic acid can be further decomposed to produce a variety of compounds,
including reducing ketones, furfural, and furan-2-carboxylic acids. In the presence of amino
acids, Vc, dehydroascorbic acid, and their degradation products undergo the Maillard
reaction, as shown in Figure 6-27.

R-CO-CO-COOH

Figure 6-26. Formation of diketo-gulonic acid through the degradation of Vc.


Vitamins 215

Figure 6-27. Vc browning pathway [16].

Figure 6-28. Relation of Vc content in cabbage and blanching time [17].

Vc has been used as natural antioxidant in foods. For example, because Vc can reduce o-
quinone compounds and thus inhibit enzymatic browning, it has been used as a bread
improver. In addition, Vc can protect folic acid and other oxidizable compounds from
oxidation. Vc is also an effective free radical scavenger. It can remove singlet oxygen,
reducing oxygen and carbon-centered radicals, and regenerate other anti-oxidants such as
copherol. The changes of Vc content during food storage in an indicator of food quality
change. Because Vc is water soluble and is sensitive to heat, pH, and oxygen, processing and
storage cause serious Vc loss in foods. Figure 6-28 shows the effects of heating time on Vc
216 Yibin Zhou, Dongfeng Wang and Ping Dong

content, Figure 6-29 indicates the retention of Vc in various processing methods, and Figure
6-30 illustrates the relationship between Vc destruction rate and aw.

Figure 6-29. Remained Vc content after various processes.

Figure 6-30. Relation between water activity and the rate of Vc destruction. O: Orange juice crystal; ●:
Sucrose solution; : Corn and soybean mixture; □: Wheat flour.

Treatment of food materials with sulfur dioxide reduces the loss of Vc. Besides, sugar
and sugar alcohols protect Vc from oxidative degradation, possibly due to their combination
with ions.
Vitamins 217

3. CHANGES OF VITAMIN CONTENTS IN FOODS


3.1. Stability of Vitamins

The contents of various vitamins in foods are affected by many factors, such as maturity
of raw materials, growth environment, soil conditions, fertilizer and water management,
illumination time and intensity, as well as post-harvest or post-slaughter handling. The
properties of the vitamins have been reported by many publications; however, only few
efforts have been made on the changes of the vitamins in complex food systems during
storage and processing. The stability of various vitamins is summarized in Table 6-12.

3.2. Effect of Maturity on Vitamin Contents

According to Table 6-13, the maturity of tomato significantly affects the content of Vc.

3.3. Change of Vitamins During Postharvest Processing and Storage

The nutrient compositions in foodstuff change significantly after harvest or slaughter.


Many enzymes, especially endoenzymes such as oxidase and hydrolase, are released from
cells.

Table 6-12 Stability of vitamins [11]

Items Light Oxidant Reductant Heat Moisture Acid Base


VA +++ +++ + ++ + ++ +
VD +++ +++ + ++ + ++ ++
VE ++ ++ + ++ + + ++
VK +++ ++ + + + + +++
VC + +++ + ++ ++ ++ +++
B1 ++ + + ++ ++ + +++
B2 +++ + ++ + + + +++
B5 + + ++ + + + +
B6 ++ + + + + ++ ++
B12 ++ + +++ ++ ++ +++ +++
Folic ++ +++ +++ + + ++ ++
acid
Biotin + + + + + ++ ++
Note: +; good stability; ++, moderate stability; +++, poor stability.
218 Yibin Zhou, Dongfeng Wang and Ping Dong

Table 6-13. Contents of Vc in tomato as a function of maturity tomato [18]

Weeks after bloom Average weight of a Color Vitamin C content (mg%)


single fruit/g
2 33.4 green 10.7
3 57.2 green 7.6
4 102.5 green-yellow 10.9
5 145.7 red-yellow 20.7
6 159.9 red 14.6
7 167.6 red 10.1

These enzymes might change the chemical structure and activity of vitamins. For
example, dephosphorylation of VB6, thiamine or riboflavin significantly affects the activities
of the vitamins
The most important factors that affect vitamin stability during storage and processing are
temperature and time, because the activities of enzymes are directly relevant to temperature
and reaction time. For example vitamin C oxidase specifically catalyzes the oxidation of
vitamin C; the loss of beta carotenes in dehydrated food is 15% after storage for 10 days and
the value reaches up to 98% after storage for 60 days.
Foods undergo multiple reactions during storage and processing and these reactions
significantly affect the sensory properties and vitamin contents of the foods. For example,
lipid oxidation produces hydrogen peroxide and epoxide, which initiate the oxidation of
carotenoid, tocopherol, and vitamin C; furthermore, hydrogen peroxide is decomposed to
carbonyl compounds, which can react with thiamine, vitamin B, and pantothenic acid and
lead to loss. In addition, carbonyl compounds generated in non-enzymatic browning of
carbohydrates also partially contribute to the loss of some vitamins. The stability of vitamins
is storage method dependent (Table 6-14).

3.4. Losses of Vitamins during Cereal Grinding

Cereals are often grinded before further processing. Grinding generates heat and hence is
a potential threat of vitamin stability. Figure 6-31 shows that effect of grinding time
(indicated by flour yield) on the retention of various vitamins.

3.5. Losses of Vitamins During Rinsing and Blanching

Rinsing and blanching causes the loss of water-soluble and heat-liable vitamins. For
example, blanching at 100°C for 2 min leads to the loss of 65% of vitamin C in cabbage and
vitamin C is completely lost after the 10 min of blanching. Steaming and boiling lead to
different vitamin losses and water-soluble vitamins are better retained in steaming than in
boiling (Table 6-15).
Vitamins 219

Table 6-14. Losses of vitamins in different storage technologies [18]

Methods Number of Loss rate (%) (1)


vegetable Vitamin A Vitamin B1 Vitamin B2 Vitamin B5 Vitamin C
speciess
Frozen 10(2) 12(4) 20 24 24 26
storage 0-50 0-61 0-45 0-56 0-78
Storage after 7(3) 10 67 42 49 51
sterilization 0-32 56-83 14-50 31-65 28-67
Note: (1) All vegetables were heated and dehydrated before storage.
(2)
Vegetables including asparagus, lima bean, kidney bean, brocoli, cauliflower, green pea, potato,
spinage, cabbage and tender corn.
(3)
Vegetables treated with hot water including asparagus, lima bean, kidney bean, green pea, potato,
spinage, and tender corn; (4) Average value.

Figure 6-31. Relationship between vitamins retention ratio and wheat flour yield [20].

Table 6-15. Retention of various vitamins in potato slices during steaming


and boiling [21]

Vitamins Boiling Steam


Vitamin C 60% 89%
Vitamin B1 88% 90%
Vitamin B5 78% 93%
Vitamin B6 77% 97%
Folic acid 66% 93%
220 Yibin Zhou, Dongfeng Wang and Ping Dong

3.6. Losses of Vitamins During Chemical Treatment

Various chemicals are often added during the storage and processing of foods and these
compounds affect the stability of some vitamins. For example, decolourants in wheat flour
causes oxidation and consequently loss of some sensitive vitamins; sulfur dioxide, sulphites,
bisulfite, and pyrosulfites are inhibitors of both enzymatic and non-enzymatic browning
reactions and protect vitamin C from oxidation.
Nitrite is added as a color fixative and preservative in bacons. This compound can react
with vitamin C and destroy thiamine and folic acid.

REFERENCES
[1] Chandler, LA; Schwartz, SJ. HPLC separation of cis-trans carotene isomers in fresh
and processed fruits and vegetables. Food Science, 1987, 52, 669-672.
[2] Pesck, CA; Warthesen, JJ. Kinetic model for photoisomerization and concomitant
photo-degradation of ß-carotenes. Journal of Agricultural and Food Chemistry, 1990,
38, 1313-1315.
[3] Dellamonica, ES; McDowell, PE. Comparison of beta-carotene content of dried carrots
prepared by three dehydrated process. Food Technology, 1965, 19, 1597-1599.
[4] Van Niekerk, PJ. Determination of vitamins, in HPLC in Food Analysis. 2nd ed. San
Diego: Academic Press, 1988.
[5] Thompson, JN; Hatina, G. Determination of tocopherols and tocotrienols in foods and
tissues by high performance liquid chromatography. Journal of Liquid
Chromatography, 1979, 2, 327-344.
[6] Herbert, V. Present Knowledge in Nutrition. 6th edition. Washington DC: International
Life Science Institute – Nutrition Foundation, 1988.
[7] Machlin, LJ. Vitamin E in commercial processing on nutrients. In Machlin LM.
Handbook of Vitamins. 2nd ed. New York: Marcel Dekker, 1991
[8] Cort, WM; Borenstein, JH. Nutrient stability of fortified cereal products. Food
Technololgy, 1976, 30, 52-62.
[9] Dennison, DJ; Kirk, J. Storage stability of thiamin and riboflavin in a dehydrated food
system. Journal of Food Processing and Preservation, 1977, 1, 43-54.
[10] Freed, MS; Brenner, S; Wodicka, VO. Prediction of thiamin and ascorbic acid stability
in canned stored foods. Food Technology, 1949, 3, 148-151.
[11] Emilia, L; Jana, K. Vitamin losses: Retention during heat treatment and continual
changes expressed by mathematical models. Journal of Food Composition and
Analysis, 2006, 19, 252–276.
[12] Woodcock, EA; Warthesen, JJ. Riboflavin photochemical degradation in pasta
measured by high performance liquid chromatography. Food Science, 1982, 47, 545-
555.
[13] Weisinger, H; Hinz, HJ. Kinetic and thermodynamic parameters for Schiff base
formation of vitamin B6 derivatives with amino acids. Archives of Biochemistry and
Biophysics, 1984, 235, 34-40.
[14] Bonjour, JP. Biotin. In: Machlin LJ, ed. Handbook of Vitamins. 2nd edition: New York:
Marcel Dekker, 1991.
Vitamins 221

[15] Flavia. NI. Lipoic acid: a unique antioxidant in the detoxification of activated oxygen
species. Plant Physiology and Biochemistry, 2002, 40, 463–470.
[16] Tannenbaum, S. Ascorbic acid. In: Fennema, O. (Ed.), Principles of Food Science Part
I Food Chemistry. Marcel Dekker, New York, 1976, 477–544.
[17] Plank, R. Handbuch der Kältetechnik. Bd. XIV. Berlin: Springer-Verlag, 1966.
[18] Maleski, W; Markakis, P. A research note. Ascorbic acid content of developing tomato
fruit. Journal of Food Science, 1971, 36, 537-539.
[19] Lund, D. Nutritional Evaluation of Food Processing. 3rd editon. New York: Van
Nostrand Reihhold, 1988.
[20] Moran, R. Biotin. Present Knowledge in Nutrition. International chemical form.
Nutrition, 1959, 122, 533-545.
[21] Zhao, H. The loss of vitamin in the process of cooking and food processing. Foreign
medical and hygiene anthology, 2003, 30, 221.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 7

MINERALS

Dongfeng Wang1, Lina Yu2, Haiyan Li1, Bin Zhang1,


Shuhui Wang3 and Xingguo Liang1
1
College of Food Science and Engineering, Qingdao, China
2
Shandong Peanut Research Institute, Qingdao, China
3
Biosystems Engineering Department, College of Agriculture - Ginn College
of Engineering, Aubum University, Auburn, AL, US

ABSTRACT
Among the 92 natural chemical elements identified to present, 81 of them have been
found in human body and 25 elements are found to be essential to life, which are termed
essential minerals. Essential minerals are widely involved in metabolisms in human body
as cofactors or activators of enzymes and some play important role in maintaining
osmotic pressure and cell membrane integrity. Besides, some minerals are important
constituents of human tissues, such as calcium and phosphorus in bone and teeth. This
chapter concerns the classification, distribution, property, and function of minerals and
their occurrence in foods. Besides, the changes of minerals in foods during harvesting,
processing, and storage are also highlighted in this chapter.

1. INTRODUCTION
1.1. Definition and Classification

To present, 115 chemical elements have been discovered, including 23 artificial and 92
natural ones. Of the 115 elements, 81 have been discovered in human bodies. According to
their importance to human body, elements are divided into 3 classes:

Essential elements

Essential elements support the normal biochemical processes of human body. Deficiency
of elements of this class leads to impaired functions of human body and the functions can be
224 Dongfeng Wang, Lina Yu, Haiyan Li et al.

restored in the early stage of the deficiency if the elements are supplemented in time.
Essential elements have special physiologic functions and cannot be replaced by other
minerals. To present, 29 elements have been found essential for the human body, including
oxygen (O), carbon (C), hydrogen (H), nitrogen (N), calcium (Ca), phosphorus (P), potassium
(K), sodium (Na), chlorine (Cl), sulfur (S), magnesium (Mg), ferrum (Fe), fluorine (F), zinc
(Zn), cuprum (Cu), vanadium (V), stannum (Sn), selenium (Se), manganese (Mn), iodine (I),
nickel (Ni), molybdenum (Mo), chromium (Cr), cobalt (Co), bromine (Br), arsenic (As),
silicon (Si), boron (B), and strontium (Sr). The former 11 elements account for up to 99.95%
of the total amount of the 29 elements in human body and are termed main or macro
elements. The remaining 18 elements are trace elements and account for only 0.05% of the
total amount.

Potential beneficial elements or supplementary elements

Elements of this class are beneficial to the physiological activities of human body when
their contents are in normal levels. However, the elements are toxic when they are present in
high levels. Rubidium (Rb), aluminum (Al), niobium (Nb), zirconium (Zr), lithium (Li), and
some rare earth elements (REEs) belong to this class.

Toxic elements

Toxic elements are also referred to as contamination elements or toxic trace elements.
These elements exert their toxicity even in very low concentrations. Bismuth (Bi), stibium
(Sb), beryllium (Be), cadmium (Cd), mercury (Hg), plumbum (Pb), and thallium (Tl) are
important toxic elements.
The elements except C, H, O, and N are termed minerals. Based on the contents in foods,
minerals are divided into main minerals, trace minerals, and ultra-trace minerals. Main
minerals include Na, K, Ca, P, Mg, S, and Cl; trace minerals include Fe, F, I, Zn, Se, Cu, Mn,
Cr, Mo, Co, and Ni; and ultra-trace minerals include Al, As, Ba, Bi, B, Br, Cd, Cs, Ge, Hg,
Li, Pb, Rb, Sb, Si, Sn, Sm, Ti, Sr, Tl, W, and V.

1.2. Distribution of Minerals in Human Body

The toxicity of main elements arranged from left to right in the same period and from top
to bottom in the same group in the element periodic table increases with their atom number
(Figure 7-1). Therefore, the argument that the effect of an element on the biological
eukaryotic cells closely relate to the periodic law of the elements is proposed. The study on
the subgroup elements shows that the change of their optimal concentration stimulated cells
growth with their abundance in the seawater is established (Figure7-2).
Figure 7-1 shows that all the 11 main elements have atom number less than 20 and they
are mainly distributed on the top of the S and P areas in the element periodic table. The
essential trace elements have the tendency to be at the position of the fourth period, especially
at the d and ds areas. Most toxic elements except Be and Ba lie in the bottom of the P area.
The majority of essential trace elements are in the top of the element periodic table and their
atom numbers are less than 35 except I and Mo. Almost all the toxic elements are located in
the lower part of the element periodic table, especially in the fifth and sixth period.
Minerals 225

The distribution of minerals in the environment and human body is displayed in Figure 7-
2. Living organisms have evolved regulatory measures to maintain essential mineral contents
in normal ranges. That is to say, a living organism can maintain a mineral in a constant range
when the element intake is insufficient or excessive.

Figure 7-1. Distribution of macro and trace elements in the element periodic table [8].

Figure7-2. Distribution of elements in the seawater (upper), human body (middle) and crust (lower).
(Data in graph are log c; the unit of c is mg/kg) [5].

Table 7-1. Functions of main minerals [4]

Mineral Functions
B Growth promotion and essential for plant growth
F Closely correlated with bone growth
Fe Component of hemoglobin, myoglobin and cytochrome
Zn Involvement in enzyme activity, nucleic acid structure and protein synthesis
I Component of thyroxine
Cu Cofactor of many metalloenzymes
Se Component of glutathione peroxidase and participation in liver function and
muscle metabolism
Mn Activation of enzyme and participation in hematopoiesis
Mo Component of molybdoenzymes
226 Dongfeng Wang, Lina Yu, Haiyan Li et al.

Table 7-1. (Continued)

Mineral Functions
Cr Effect of insulin intensive and glucose utilization promotion
Mg Activation of enzyme and component of bone
Si Participation in skeletogeny
P Component of ATP
Co Component of VB12
Ca Component of bone and involvement in neural transmission
S Component of proteins
K Electrochemical and messenger function and extracellular cations
Na Electrochemical and messenger function and extracellular cations
Cl Electrochemical and messenger function and extracellular anions

1.3. Functions of Minerals in the Living Body

Table 7-1 lists the functions of main minerals for living organisms.
Minerals play important roles in the human body and their functions often involve
complex mechanisms. The functions of the minerals are affected by their interactions (such as
coordination and antagonism) with other food constituents and the compounds in human
body. For example, P and Ca mutually affect their absorption and their absorption rates are
the highest when the two minerals are present in molar ratio 1:1 in foods. Fe increasingly
antagonizes the absorption of Zn in Fe/Zn ratios range from 1:1 to 22:1 in diets and Zn
reduces the absorption of Cu. Many researches have revealed that both deficiency and
imbalance of minerals cause a number of abnormalities in living organisms.
In contrast to most organic constituents, minerals cannot be synthesized in the human
body and must be ingested from foods and drinking water. Besides, minerals cannot be
metabolized and are repelled from the body by excreting. Therefore, dietary structure
significantly influences the contents of minerals and their proportions in the body.

2. THE EXISTENCE STATES OF MINERALS IN FOODS


Minerals in foods can be present in the dissolved or non-dissolved, colloidal or non-
colloidal, organic or inorganic state, ionic or non-ionic state, or complexed and non-
complexed state.
The existence states of minerals significantly influence their bioavailability and the safety
of foods. For instance, iron in heme is absorbed much easier than other iron forms and its
bioavailability is much less affected by other factors in diets, including iron absorption
inhibitors. The absorption of minerals in foods is also affected by the presence of other food
components. For example, vitamin C enhances the absorption of iron, but phytic acid and
polyphenols inhibit iron absorption.
The toxicity of toxic elements, such as Cd, Pb, and Hg, is affected by the presence other
constituents in foods. For example, vitamin C decreases the toxicity of Cr6+ by reducing it to
Cr3+; phytic acid, polyphenols, and proteins reduce the toxicity of Pb and Cd by complexing
with them. The toxicity of heavy metals can be antagonized by other minerals. Zinc is the
Minerals 227

metabolic antagonist of Cd, competing with Cd for the thiol of metallothionein. Pb is more
poisonous in the absence of Fe and Cr in the diet; Se can complex with Hg and reduce Hg
toxicity. The valence significantly influences the toxicity of metals. As3+ is much more toxic
than As5+; methyl arsenic acid and dimethyl arsenic acid are moderately toxic, while
arsenobetaine and arsenocholine are almost non-toxic. Cr3+ is an essential trace element,
while Cr6+ is toxic and Cr2O72- is a strong carcinogen.
Hence, in evaluating the bioavailability and safety of a mineral in foods, its existence
state must be taken into consideration. Generally, only very few minerals are present as free
ions in foods.

2.1. Combination with Monosaccharides and Amino Acids

According to the Lewis acid-base theory, metals are Lewis acids which can provide
empty orbitals. Small molecules that are rich in N, S, and O atoms, such as sugars, amino
acids, nucleic acids, chlorophylls, and hemoglobins, contain lone pair electrons and can act as
Lewis bases. Therefore, minerals can form complexes with above-mentioned biomolecules.
α-Amino acids are bidentate ligands and bond to metal ions via an oxygen of the
carboxylic acid group and the nitrogen of the amino group to form a five-membered ring, as
shown in Figure 7-3. Under certain conditions, the side chains of amino acids also participate
in the coordination. In addition to carboxyl terminal peptide and certain groups of amino acid
side chains, the carboxyl in peptide and imino groups are also involved in coordination [10].

O O

C H
N CH2
H2 C Zn
C
N O
H O
Note: Left: Zn(Gly)22H2O, Midmost Glycine tripeptide mineral complex, M = Cu (II) or Ni (II); Right:
Glycine tetrapeptide cupric complex.

Figure 7-3. Structures of mineral-peptide complexes.

Both plant- and animal-derived foods contain a large number of monosaccharides,


carbohydrate derivatives and amino acids. As long as the adjacent hydroxyls in sugars are
arranged in favorable spatial configurations, coordination with metals occur readily. For
example, the three hydroxyl groups on pyranose in the axial-transverse-axial configuration
and the three hydroxyl groups on furanose in the cis-cis-cis structure favor the coordination
with divalent or trivalent minerals. The presence of carboxyl and imino groups in
carbohydrates increases the stability of their complexes with minerals by several orders of
magnitude. Yunlan Su, et al. determined the molecular structure of the lanthanum-galactose
acid complex by infrared spectroscopy and found two complexes with different structures.
Iron can generate multi-core complexes with glucose, fructose and other monosaccharides.
The osamines generated during the Maillard reaction, such as glycosyl amines and
glucosyl amines, can react with minerals to form stable complexes. Nagy L [6] et al found
that the Amadori isomer formed during the Maillard reaction between glucose and amino
acids at high temperatures is a more stable mineral ligand.
Structural analysis of complexes formed between Cu(II) and aminosaccharide reveals that
all the aminosaccharides are the binary ligands, in which -NH2 is the main coordination group
228 Dongfeng Wang, Lina Yu, Haiyan Li et al.

for Cu(II) and other divalent minerals and the -OH group plays less role in coordination with
Cu(II). The –OH group on 1-C of GlcNH2GalNH2 is the main coordination group for divalent
minerals, but if the -OH on 1-C is not available due to steric hindrance, such as that in
aminosaccharide derivatives, other –OH groups participate in the coordination. Nagy L et al.
[6] studied the structural parameters of carbohydrate-mineral complexes by EXAFS
(extended X-ray absorption fine structure spectroscopic) and the results are given in Table 7-
2. It could be seen that minerals in foods can form various complexes with carbohydrates and
their derivatives.

Table 7-2. Structural parameters of carbohydrate-mineral complexes

Ligands Binding Interaction N γ/pm σ/pm


Fru Fe-O 6 195 9.8
Fe…C 4 277 7.9
Fe…Fe 1 310 7.0
Rib Cu-O(eq) 4 191 7.5
Cu-O(ax) 2 230 13.0
Cu…C 2 271 10.0
GlcNH2 Cu-O(eq) 3 193 7.7
Cu-O(eq) 1 193 7.7
Cu-O(ax) 2 234 13.0
Cu…C 2 274 10.0
Adenosine Cu-O(eq) 4 191 5.4
Cu-O(ax) 2 232 7.8
Cu…C 2 275 8.4
Uridine Cu-O(eq) 4 192 7.9
Cu-O(ax) 2 234 7.9
Cu…C 2 279 9.6
HyA Cu-O(eq) 4 191 8.2
Cu-O(ax) 2 234 3.2
Cu…C 2 313 13.0
Zn-O 4 202 8.1
Zn…C 2 299 13.2
N-D-glugly Cu-O(eq) 2 190 5.6
Cu-N(eq) 2 190 5.6
Cu-O(ax) 2 215 2.5
Cu…C 4 270 2.8
Cu…Cu 1 297 11.4
N-D-glugly Ni-O,N 6 204 7.6
Ni…C 6 285 5.4
N-D-glugly Co-O,N 6 200 11.9
Co…C 6 290 11.7
Co…Co 1 303 7.6
PHTAc Zn-O,N 4 205 8.0
Zn…C 6 290 10.0
Zn…O,C,S 6 286 14.0
PHTAc Mn-O,N 6 216 9.5
Mn…C 6 305 11.0
Mn…O,C,S 8 375 18.0
Minerals 229

Ligands Binding Interaction N γ/pm σ/pm


PHTAc Ag-N 1 203 5.0
Ag-S 1 230 4.6
Ag…C 4 294 19.0
PHTAc Ni-O,N 6 203 8.0
Ni…C 6 284 9.5
Ni…O,C,S 8 390 15.0
LacA Mn(IV)-O 6 208 2.3
Mal Mn(IV)-O 6 208 3.5
GlcA Mn(IV)-O 6 209 1.9
Sacc Mn(III)-O 6 209 3.6
Mal Mn(III)-O 6 206 1.0
GlcA Mn(III)-O 6 208 3.3
Note: N, coordination number; γ, atomic distance; σ, Debye-Waller factor; Fru, D- Fructose; Rib, D-
Rhamnose; GlcNH2, 2-amino-2-deoxy-D-Glucose; HyA, Hyaluronic Acid; N-D-glugly(N-D-
gluconylglycine, a pseudopeptide derivative glucono-delta-lactone and glycine); PHTAc, [2-
(polyhydroxyalkyl)thiazolidine-4-carboxylic acid]; LacA, D-lactobionic acid; Mal, maltitol 4-O-ζ-
D-glucopyranosyl-D-glucitol; GlcA, D- Gluconate; Sacc, D- Sucrose; Ara, L- or D- Arabinose.

2.2. Combination with Oxalic Acid and Phytic Acid

Oxalic acid in plant-derived foods is an important mineral chelator. When oxalic acid and
phytic acid are present in high levels in foods, the bioavailability of some essential minerals
and the toxicity of some toxic elements will be significantly reduced.
Phytic acid can form insoluble complexes with Ca, Fe, Mg, Zn and many other minerals.
Besides, phytic acid is also able to form ternary complexes with proteins and minerals. The
reaction not only reduces the bioavailability of proteins, but also weakens the absorption of
minerals. About 10% of P in vegetables cannot be absorbed by bodies because of its
combination with phytic acid. In cereals, the percentage is as high as about 40% and even up
to 90% in some species, as shown in Table 7-3.

2.3. Combination with Nucleotides

The biological functions of certain nucleotides are closely related to minerals. For
example, in the presence of Mg, ATP can be hydrolyzed to ADP and phosphate. All the three
components in nucleotides have the ability to coordinate with minerals and their coordination
capacities decrease in the following order: bases > phosphate groups > pentose. Bases
coordinate with minerals through N-3 of pyrimidine and N-7 of purine base. Ca2+, Mg2+,
Cu2+, Mn2+, Ni2+ and Zn2+ are common coordination minerals for nucleotides. In the case of
coordination with ATP, Ca2+ and Mg2+ bond only to the phosphate groups, whereas Cu2+,
Mn2+, Ni2+ and Zn2+ combine with both phosphate groups and N-7 of adenine. The stability
constants of ATP-divalent minerals follow the following order: Cu2+ > Zn2+ > Co2+ > Mn2+ >
Mg2+ > Ca2+ > Sr2+ > Ba2+ > Ni2+. NMR and Raman spectroscopy reveal that the molar ratio
of the Mg2+-ATP complex is 1:1 (Figure 7-4). 1H-NMR and 31P-NMR indicate that Cu2+
binds to the N-7 of purine or the N-3 of pyrimidine on ATP.
Cu2+, Co2+, Ni2+ and Cd2+ coordinate with the phosphate and N-7 of adenine in ATP in
two ways, with the resultant structures termed macrochelated inner sphere (Figure 7-5A) and
230 Dongfeng Wang, Lina Yu, Haiyan Li et al.

macrochelated outer sphere (Figure 7-5B) respectively. In the former structure, N-7 of
adenine directly binds to the mineral, but the α-phosphate group coordinates through H2O. In
the later structure, N-7 of adenine coordinates with mineral through H2O, whereas the α, β, γ-
phosphate groups directly link with the mineral (Figure 7-5).

Table 7-3. Content of phosphorus coordinated with phytic acid in


plant food

Food Phosphorus coordinated with Food Phosphorus coordinated with


Phytic Acid Phytic Acid
mg/100g % mg/100g %
Oat 208~355 50~88 Potato 14 35
Wheat 170~280 47~86 Kidney Bean 12 10
Barley 70~300 32~80 Carrot 0~4 0~1
Rye 247 72 Orange 295 91
Rice 157~240 68 Lemon 120 81
Corn 146~353 52~97 Walnut 120 24
Peanut 205 57 Soybean 231~575 52~68

Figure 7-4. Complex of Mg with ATP [4].

a)

Figure 7-5 (continued)


Minerals 231

b)

Figure 7-5. Two simplified structural models of (ATP) 2-Chelate inner ring (A) and external ring (B)
coordination sphere (M indicates a mineral ion) [7].

2.4. Combination with Ring Ligands

Minerals can complex with planar ring ligands in addition to low molecular weight
carbohydrates, amino acids and peptides mentioned above. Take porphyrins as example.
Porphyrins are the derivatives of porphine consisting of four pyrrole rings connected by four
carbon atoms (Figure 7-6a, b). When H atoms in numbered positions of the porphine ring are
replaced by other groups, porphyrins are generated (Table 7-4).

a)

b)

Figure 7-6. Structure of porphine. In (a) and (b), the structure is numbered in
different ways.
232 Dongfeng Wang, Lina Yu, Haiyan Li et al.

Table 7-4. Important Porphyrins

Porphyrins Substituents
1 2 3 4 5 6 7
Protoporphyrin IX M V M V M P P
Mesoporphyrin IX M E M E M P P
Deuteroporphyrin IX M H M H M P M
Hematoporphyrin IX M B M B M P P
Spirograhic porphyrin IX M F M V M P P
Corproporphyrin III M P M P M P P
Aetioporphyrin III M E M E M E E
Uroporphyrin III A P A P A P P
Note: A=-CH2COOH, B=-CH (OH) CH3, E=-C2H5, F=-CHO, M=-CH3, H=-H, P=-CH2CH2COOH, V=-
CH=CH2.

Porphyrins can form complexes with Fe2+, Fe3+, Zn2+, Co2+, Cu2+, Mg2+ and other mineral
ions, such as Fe in hemoglobin and Mn in chlorophyll.
Heme (Figure 7-7) consists of an iron atom and a planar porphyrin ring. The iron in the
center is connected with the N-atoms of four pyrrole rings through coordination bonds, and
the fifth coordination point is provided by a histidine residue in globin, and the remaining
sixth coordination point comes from negatively charged atoms in other ligands, such as H2O.
Heme can connect with globin to form myoglobin (Figure 7-8).
Three types of hemes, namely heme a, heme b and heme c, have been found. Heme a is
the prosthetic group of cytochrome c oxidase; heme b is an iron-protoporphyrin IX complex
(Table 7-4) and it is the prosthetic group of hemoglobin, myoglobin, cytochrome b,
cytochrome p-450, catalase and peroxidase; heme c is the prosthetic group of cytochrome c.

Figure 7-7. Structure of heme.

Chlorophyll is a green pigment in the higher plants and photosynthetic organisms. It has a
planar molecule and consists of four pyrrole rings connected through the methylene bridge.
The Mg in center is connected with the N-atoms of four pyrrole rings through coordination
bonds (Figure 7-9). Many chlorophyll species have been identified, such as chlorophyll a, b, c
and d, as well as bacteria chlorophyll and chlorobium chlorophyll. Chlorophyll a and b occur
widely in plant-derived foods in the ratio 3:1. Chlorophyll is transformed to pheophytin
during food processing. In acidic conditions, the Mg atom in chlorophyll is replaced by
Minerals 233

hydrogen atoms and the resultant pheophytin is dark olive-brown. Heating accelerates the
transformation. In addition to hydrogen, Mg can also be replaced by other divalent minerals,
such as zinc and copper.

Figure 7-8. Structure of myoglobin.

Figure 7-9. Structure of chlorophyll.

Vitamin B12 consists of a corrin ring and has four pyrrole subunits. Because cobalt is
essential for its bioactivity, Vitamin B12 is also known as cobalamin. Vitamin B12 is a red
crystal and its systematic name is α-(5,6-dimethylbenzimidazolyl)cobamidcyanide. In the
structure, the cobalt atom combines with four inner nitrogen atoms in the corrin ring. If the
sixth coordination site is replaced by cyanide, VB12 is transformed to cyanocobalamin; if the
cyanogroup connected with cobalt is replaced by the hydroxyl group, VB12 is converted to
hydroxocobalamin, which is the ubiquitous form of VB12 in nature. If the cyano group is
replaced by the nitroso group, nitriocobalamin is generated. Nitriocobalamin is found mainly
in some bacteria. The structure of Vitamin B12 is given in Figure 7-10.

2.5. Combination with Proteins

Protein consists of amino acids. In addition to peptide bonds, terminal amino groups and
carboxyl groups, as well as some side groups of amino acid residues, such as the hydroxyl
group in serine and threonine, the phenolic hydroxyl group in tyrosine, the carboxyl group in
234 Dongfeng Wang, Lina Yu, Haiyan Li et al.

acidic amino acids, the amino group in basic amino acids, the imidazolyl group in histidine,
the sulfhydryl group in cysteine, and the thioether group in methionine, can coordinate with
minerals. However, only the groups in suitable sites can form complexes with minerals.

Figure 7-10. Structure of vitamin B12.

Figure 7-13. schematic diagram of Zn2+ in carboxypeptidase A [3].

Figure 7-11 is the schematic diagram of carboxypeptidase A. Zn2+ coordinates with the
protein constituent through two imidazole groups and a carboxyl group in the protein.
Metals are involved in the activity of a large number of enzymes and such enzymes are
known as metalloenzymes. Zn, Fe, Cu, Mn, Mg, Mo, Co, K, and Ba are the most important
cofactors for such enzymes.
Living organisms synthesize a large variety of proteins for the storage and transport of
minerals.
Minerals 235

Ferritin

Ferritin is mainly distributed in the spleen, liver and bone marrow of animals, as well as
in plants chloroplasts and certain fungi. Its main physiological function is to store unused or
excessively absorbed iron.

Transferrin

Transferrin is mainly distributed in body fluids and cells of vertebrates. A large variety of
transferrins have been identified, such as serotransferrin in serum and lactotransferrin in milk
and lacrimal gland secretion. Serotransferrin is a class of mineral-binding glycoproteins with
molecular weight about 7.7~7.4 × 104. After the transferrin releases all Fe3+ irons, the protein
constituent can combine with other divalent or trivalent ions other than iron, such as Cu2+,
Zn2+, Cr3+, Mn3+, Co3+, and Ga3+.

Iron-sulphur protein

Iron-sulphur proteins are characterized by the presence of the Fe-S chromophore and
their molecular weight is about 10 kD. All the Fe atoms in the proteins have variable valence.
The proteins participate in redox reactions in organism as the electron transfer group and are
involved in biological oxidation, nitrogen fixation and photosynthesis.

Cuprein

Cupreins are copper-containing proteins and superoxide dismutase is a typical example of


such proteins. To present, more than forty kinds of cupreins have been found. Cupreins with
blue color are called blue copper proteins and those without the blue color are called non-
ceruloplasmins.

Minerallothionein

Metallothionein (MT) was firstly separated from horse kidney in 1957. Later, it was
found that MT is widespread in nearly all organisms. MTs are a category of induced protein
with a molecular weight about 6~10 kD. MTs are characterized by their high Cys contents,
reaching up to 25~35%. The functions of MTs include anti-oxidation, free radicals
scavenging, heavy metal complexing, and mineral storage. The structures of MTs are highly
conserved. Many minerals, such as Cd, Pb and Zn, can induce the synthesis of MTs.
One of the most important functions of MTs is to the alleviate toxicity of heavy metals.
The affinity of MTs to some heavy metals follow the order Cd2+ > Pb2+ > Zn2+. Because the
MT level is positively correlated with contaminant level, it is widely used as an indicator of
environmental pollution.
Capasso, C. et al [1] determined the three-dimensional structure of MT-nc obtained from
the Antarctic fish (Notothenia coriiceps). NMR analysis showed that MT-nc is composed of
the α- and β- domains. The α-domain in the C-terminal consists of eleven Cys residues and
four mineral atoms, and the β-domain in the N-terminal has nine Cys residues and three
mineral atoms (Figure 7-12).
The ninth cystine of MT-nc in the α domain is different from that of mammals.
236 Dongfeng Wang, Lina Yu, Haiyan Li et al.

The terminal amino acid sequence of mammal-derived MTs is CXCC (C denotes Cys, X
denotes any amino acid), while the sequence of fish MT is CXXXCC. This difference
attributes to the different structures of their α domains.
Except few exceptions, the N-terminals of mammal-derived MTs are acetyl Met, and the
carboxyl-terminals are Ala. Each MT molecule contains 20 Cys residues and the Cys residues
are in the same positions. The Cys residues can form five Cys-X-Cys units, one Cys-Cys-X-
Cys-Cys unit and one Cys-X-Cys-Cys unit.
Although the amino acid compositions of MTs in different animal are quite similar, the
MTs differ in the metal binding capacity. Generally, each MT molecule can bind seven
mineral ions and the affinities of some metals to MTs follow the increasing order Zn2+ < Pb2+
< Cd2+ < Cu2+ < Ag+ < Hg+.

a)

b)

a
The main chain composite structure of 31-60 residues in α domain;
b
The main chain composite structure of 2-28 residues in β domain.

Figure 7-12. NMR diagram of MT-nc.

Phytochelatins

Phytochelatins are firstly found in plants and have similar metal binding capabilities as
MTs. It is estimated that more than 90% of Cd2+ in plant cells are bound with phytochelatins.
The general formula of phytochelatins is (γ-Glu-Cys)n-Gly. Phytochelatins are major
heavy metal binding peptides in plants and some yeast species. The biosynthesis of
phytochelatins is induced rapidly after exposure to heavy metals, especially Cd2+ and Hg2+.
Heavy metals in living organism are present in both the soluble and insoluble forms, in
which, soluble heavy metals bind mainly to phytochelatins. Phytochelatins from different
crops have the following common characteristics: (1) the molecules have similar structure, as
Minerals 237

shown in Figure 7-14; (2) the Glu residue locates in the N-terminal; (3) the amino acid
connected with Glu-γ-oxo is Cys; (4) the γ-Glu-Cys unit occurs repeatedly in the structure.

Figure 7-14. Structure diagram of phytochelatins.

2.6. Complexation with Polysaccharides

Polysaccharides can complex with metals through thiol, amino, and carboxyl groups in
addition to hydroxyls. Complexation with metals enhances the biological functions of
polysaccharides and provides an effective way for removing harmful minerals.
Polysaccharide chains contain a large number of groups for coordination with metals,
such as pectin and alginate of the plated ribbon-type conformation (Figure 7-15). The
conformation of polysaccharides significantly affects the metal-binding capability. In the case
of alginate, Ca2+ facilitates the maintenance of the egg-box conformation, as shown in Figure
7-16.

Figure 7-15. Plated ribbon-type conformation of pectin and alginate.

Ca Ca Ca Ca

Figure 7-16. Sketch map of egg box conformation.


238 Dongfeng Wang, Lina Yu, Haiyan Li et al.

3. PHYSICOCHEMICAL PROPERTIES, TROPHISM AND SAFETY OF


MINERALS IN FOOD
3.1. Physicochemical Properties

3.1.1. Solubility
The transfer and metabolism of most minerals in all biological organisms occur in
aqueous solutions. Therefore, the bioavailability and activity of minerals in foods, to a great
extent, depend on their solubility in water. Mg, Ca and Ba are in the same group (II A) and
their halogenides are readily soluble in water. However, their hydroxids, carbonates,
phosphates, sulphates, oxalates and phytates, are sparingly soluble. Besides, the presence of
phytic acid significantly reduces the bioavailability of some minerals, such as Fe, Zn, Ca, Mg
and Mn.
The solubility of minerals is influenced by pH and other constituents in foods. Generally,
minerals are present in the dissolved state in pH below 6 and their complexation with
proteins, amino acids, organic acids, nucleic acids, nucleotides, peptides and carbohydrates
markedly improve their solubility and bioavailability. Harmful minerals can be removed by
the formation of insoluble complexes. For example, citric acid is sometimes used as a chelator
in lead poisoning treatment.

3.1.2. Oxidability and Reducibility


Minerals might occur in multiple valences in foods and their valences are subject to
changes due to the presence of other food components. The valence of a metal largely affects
its significance for human body. For example, Fe2+ is beneficial for the biological process of
living organisms, but Fe3+ are toxic; Cr2+ and Cr3+ are not toxic or only moderately toxic, but
Cr6+ is a well-known strong carcinogen (LD50 6~8 g). Cr6+ in blood stream can be oxidized by
oxygen to chromium oxide and transform hemoglobin to methemoglobin, leading to
decreased oxygen level in blood. Inhale of a small amount of chromic salt or chromic acid
(+6) cause great pathological changes in the kidney, liver, nervous and blood.

3.1.3. Activity
The reactivity of an ion in biochemical reactions depends on its activity instead of its
concentration. The definition of the activity of minerals is as follows:

ai= fi·Ci

where: ai denotes the activity of the ion, fi denotes the activity coefficient, and Ci denotes the
ion concentration.
The activity coefficient (fi) increases as the ion concentration decreases. No method has
been developed to determine the accurate activity of minerals in foods.
Because the ion activity is positively correlated with ion concentration, the ion activity
can be presented by ion concentration. Indeed, all mineral-involved biochemical reactions are
related to the concentrations, states, valence state, dietary patterns, and so on (Figure 7-18).
Minerals 239

Figure 7-18. Schematic relationship between biological activity and relative concentration of minerals.

3.1.4. Chelating Effect


Most metal ions in foods are present as complexes by coordinating with organic
molecules. According to Werner‘s coordination theory, coordination compounds are divided
into two parts. Take [Cu(NH3)4]SO4 as an example. Cu(NH3)42+ is defined as the internal
limiting ion, Cu as the central ion, NH3 as the ligand, and SO42- as the external ion. In foods,
most internal limiting ions are transition elements, such as Fe, Co, Mg, and Cu. The central
ions are connected to certain atoms on ligands, such as O, N and S, which are called
coordination atoms. C, H, VA group elements, and VIA group elements are important
coordination atoms.
A chelate complex is characterized by the presence of two or more separate coordinate
bonds between a polydentate (multiple bonded) ligand and a single central atom. Chelate
complexes are contrasted with coordination complexes composed of monodentate ligands,
which form only one bond with the central atom. Many minerals exert their functions by
chelating or coordination with organic ligands. For example, Fe in heme is essential for
oxygen transport and Mg in chlorophyll enables photosynthesis. In addition, many metals,
such as Fe2+, Mg2+, Co2+, Mo2+, Mn2+, Cu2+ and Ca2+, can coordinate with specific groups on
the side chains of amino acids in metalloenzymes.

3.2. Nutrition and Toxicity

3.2.1. Nutrition
Atomic absorption spectrophotometry is a conventional method for determining the total
content of a mineral in foods. However, the bioavailability and nutritional value of minerals
are affected by multiple factors and the information on the total content is insufficient for
bioavailability and nutrition evaluation. The utilization of minerals is discussed below by
taking Fe as an example.
Fe in diets is present in two forms: heme-iron and non-heme iron. Heme-iron occurs
primarily in animal-derived foods. The heme-iron is bivalent and can be absorbed directly and
stored in intestinal ferritin for human utilization. Non-heme iron is the major form of Fe in
plant-derived foods, such as cereals food, vegetables, fruits and legumes. Non-heme iron is
trivalent and cannot be easily absorbed unless it is reduced to its bivalent form. Some plant-
240 Dongfeng Wang, Lina Yu, Haiyan Li et al.

derived foods contain high contents of phosphates, oxalic acid and tannic acids and these
compounds can precipitate Fe by complexing with it, which further reduces Fe utilization.
The presence of reducing agents in plant-derived foods improves Fe utilization. Generally, the
bioavailability of Fe in animal-derived foods is higher than that in plant-derived foods, as
shown in Figure 7-19.
The utilization of Fe is associated with its sources, existence state and diet structure. For
example, phosphoric acid in milk can precipitate Fe and reduces its utilization; polyphenols in
tea can complex with Fe and affect its utilization; diet with insufficient Cu decreases Fe
absorption, because Cu is essential for hemoglobin synthesis.
The absorption of Fe is also affected by individual or physiological factors. People
suffering Fe deficiency or anemia absorb Fe easily and women generally have a better
absorption than men. Besides, the possibility of Fe deficiency decreases with age increase.

3.2.2. Toxicity
All minerals are toxic when they are present in high levels in foods. In addition to
contents, the toxicity of minerals is also affected by the following factors.

Figure 7-19. Utilization of Fe element in different sources. 1 to 12 represents rice, spinach, beans, corn,
lettuce, wheat, soybean, iron, protein, bovine, fish, hemoglobin, and beef, respectively.

Synergistic or antagonistic actions between minerals

The presence of a metal might enhance or decrease the toxicity of another. For example,
As increases the toxicity of Pb; Cu increases the toxicity of Hg; Cu decreases the toxicity of
Hg; Mo significantly reduces Cu absorption; Co increases the toxicity of S; Se decreases the
toxicity of Cd and Ni; Cd antagonizes the absorption of Zn and Cu.

Valence

The toxicity of an element is closely related its valence. The toxicity of elements depends
on their coordination with biological macromolecules and the valence markedly influences
the coordination. For example, Cr3+ is an essential trace element, while the Cr6+ is highly
toxic.

Chemical form

The toxicity of an element is also related to its chemical forms. For example, the LD50s
(mg/kg) of various arsenides are arsenite 14.0, arsenate 20.0, methyl arsenate 700~1800, 2-
Minerals 241

methyl arsenate 700~2600, arsenocholine 6500 and betaine arsenate complex >10,000. It
could be seen that volatile inorganic arsenic is the most toxic. Arsenocholine and betaine
arsenate are often considered non-toxic.

Poisoning mechanism of heavy metals

The poisoning mechanisms of heavy metals are rather complex and affected not only by
their contents in the body, but also by the exposing pathway, metabolism, and health
conditions of ingesters. In general, heavy metals exert their toxicity in one or more of the
following mechanisms:

Heavy metals disrupt the functional groups in biomolecules. For example, Hg(II)
and Ag(II) can bind with –SH of Cys residues in enzymes and block the
participation of –SH in enzymatic reactions.
Heavy metals replace the essential metals in biomolecules. The activity of many
enzymes is closely related to the metal cofactors. The activity of metalloenzymes is
easily destroyed by the replacement of metal factors. For example, Be(II) can
replace the Mg(II) in Mg(II)-containing enzymes.
Heavy metals change the conformation of biomolecules. Heavy metals can bind to
various biomolecules, such as proteins, nucleic acids and change their
conformations. For example, Pb2+, Cd2+ and Ni2+ can bind to double stranded DNA
(dsDNA) and this interaction leads to different modifications in the dsDNA
structure.

3.2.3. Effect of Existence State


The existence states of minerals tremendously influence their nutrition and toxicity.

The existence state determines mineral solubility.

Minerals must be in the dissolved state for their nutrition or toxicity. Minerals often occur
as complexes with various organic ligands in foods and the ligands tremendously influence
mineral solubility and bioavailability. For example, Ca in protein-Ca complexes is absorbed
easily, but that in the Ca-oxalic acid complex is insoluble and possibly causes urinary
calculus. The solubility of minerals also depends on the counter ion. For a same heavy metal,
its oxide is less poisonous than its soluble chloride and nitrate. Generally, the counter ion
influences the toxicity of a metal in the following order: Nitrate > Chloride > bromide >
acetate > iodide > perchlorate > sulfate >> phosphate > carbonate > fluoride > hydroxide >
oxides. The solubility of metal salts in aqueous solution decreases with the increase of atomic
weight of the metals.

The existence states decide the physiological functions of minerals.

The physiological functions of minerals are closely related with their valences. Cr3+ plays
an important role in glucose, fat, and cholesterol metabolisms. In contrast, Cr6+ is highly
toxicity and its in vivo transformation to Cr3+ is nearly negligible. When the body
accumulates an excessive amount of Cr6+, poisoning symptoms appear.
242 Dongfeng Wang, Lina Yu, Haiyan Li et al.

The coexistence of other minerals changes the nutrition and toxicity a metal.

Hg, Pb and Cd are well-known toxic metals, but the intake of certain components may
strengthen or weaken their toxicity. For example, lipopolysaccharides increase the
accumulation of Hg in mice and augment the Hg-induced nephrotoxicity.

Table 7-5. Minerals contents in some foods [2, 9]

Content / mg/100g
Food Ca Mg P Na K Fe Zn Cu Se
Fried eggs 57 13 269 290 138 2.1 2.0 0.06 8
Wheat bread 35 6 30 144 31 0.8 0.2 0.04 8
Graham bread 20 26 74 180 50 1.5 1.0 0.10 16
Salt-free macaroni 5 13 38 1 22 1.0 0.4 0.07 19.0
Cooked rice 10 42 81 5 42 0.4 0.6 0.01 13.0
Instant rice 10 42 81 5 42 0.4 0.6 0.01 13.0
Mature black soybean 24 61 120 1 305 2.0 1.0 0.18 7.9
Red cashew 25 40 126 2 356 3.0 0.9 0.21 1.9
Whole milk 291 33 228 120 370 0.1 0.9 0.05 3.0
Skim milk /nonfat milk 302 28 247 126 406 0.1 0.9 0.05 7.6
Ameican cheese 261 10 316 608 69 0.2 1.3 0.01 3.8
Sayda cheese 305 12 219 264 42 0.3 1.3 0.01 7.0
Farmhouse cheese 63 6 139 425 89 0.1 0.4 0.03 7.3
Low fat yogurt 415 10 326 150 531 0.2 2.0 0.10 5.5
Vanilla ice-cream 88 9 67 58 128 0.1 0.7 0.01 4.7
Overbark baked potato 20 55 115 16 844 2.8 0.7 0.62 1.8
Underbark boiling otato 10 26 54 7 443 0.4 0.4 0.23 1.2
Palm cabbage, crude stem 216 114 297 123 1470 4.0 2.0 0.40 0.9
Palm cabbage, cooked stem 249 130 318 141 1575 4.5 2.1 0.23 1.1
Crude shattering carrot 15 8 24 19 178 0.3 0.1 0.03 0.8
Cooked freezing carrot 21 7 19 43 115 0.4 0.2 0.05 0.9
Fresh entire tomato 6 14 30 11 273 0.6 0.1 0.09 0.6
Canned tomato juice 17 20 35 661 403 1.0 0.3 0.18 0.4
Attains (defrosting) 17 18 30 2 356 0.2 0.1 0.08 0.4
Attains 52 13 18 0 237 0.1 0.1 0.06 1.2
Apple (skin) 10 6 10 1 159 0.3 0.1 0.06 0.6
Banana (peeling) 7 32 22 1 451 0.4 0.2 0.12 1.1
Roasted beef (cans) 5 21 176 50 305 1.6 3.7 0.08 —
Roasted veal (cans) 6 28 234 68 389 0.9 3.0 0.13 —
Roasted chicken breast 13 25 194 62 218 0.9 0.8 0.04 —
Roasted chicken drumstick 10 20 156 77 206 1.1 2.4 0.07 —
Cooked salmon 6 26 234 56 319 0.5 0.4 0.06 —
Canned salmon with bone 203 25 277 458 231 0.9 0.9 0.07 —
Minerals 243

4. CONTENTS OF MINERALS IN FOODS


The contents of minerals in foods are governed by multiple factors, such as place of
production, cultivation methods, and processing techniques. For example, the Cu content in
rice is influenced by the Cu content in soil, climate, water source, use of fertilizer, insecticide,
pesticide and fungicide, and processing equipment. The mineral contents of some foods are
shown in Table 7-5. Besides, the utilization of minerals in foods is closely associated with the
dietary structure due to the interactions with other food components. This section concerns
only the impact of material, processing, and storage on mineral contents in foods.

4.1. Effect of Raw Material on Mineral Contents in Foods

The contents of minerals in plant-derived foods are subject to the influences of soil, water
and fertilizer management, antagonistic effect among minerals and climate. As shown in
Table 7-6, the contents of some minerals in black glutinous rice of three different provinces in
China differ greatly.
The contents of minerals in animal-derived foods depend on feed, animal health status
and environment, in which, the mineral content of feed is the most important determinant. As
shown in Table 7-7, the supplementation of trace elements in feed markedly increases the
contents of K, Ca, and P in milk.

4.2. Effect of Processing on Mineral Contents in Foods

Processing method, water used, processing equipment, and food additives also change the
contents of minerals in foods.
Table 7-8 lists the effects of four processing methods on the mineral contents of instant
fiddlehead. It could be seen that the four treatments slightly increase the contents of Ca, but
more or less decrease the contents of other minerals, of which, salting-out dehydration plus
blanching leads to the highest loss of Mg, Mn, and Zn, and salting-out dehydration without
blanching causes most Fe and Cu losses.
Table 7-9 illustrates the loss of some minerals in spinach after blanching. K and Na suffer
the highest loss after blanching, but the Ca content is nearly not affected. Hence, minerals in
free state, such as K and Na, are lost easily during bleaching, but minerals occuring in
insoluble state (such as Ca) are not lost.

Table 7-6. Contents of some minerals in black glutinous rice sampled from three
provinces in China (mg/100g)

producing area Zn Cu Fe Mn Ca Mg
Hunan 19.48 1.779 17.18 15.46 27.59 12.27
Zhejiang 19.47 2.549 20.13 24.25 59.48 12.00
Guizhou 17.64 0.702 24.97 25.36 32.00 11.42
244 Dongfeng Wang, Lina Yu, Haiyan Li et al.

Table 7-7. Effect of trace element supplementation in cattle feed on the minerals content
of milk (mg/100g)

Fe Cu Zn Mn K Na Ca Mg P
Experimental group 0.122 0.032 0.417 0.008 81.60 83.70 144.0 11.00 98.60
Control group 0.137 0.007 0.442 0.010 68.39 85.34 77.67 10.06 82.09

Table 7-8. Effect of different processing methods on some trace elements content in
instant fiddlehead (mg/100g dry weight)

Processing methods Ca Mg Fe Mn Cu Zn
Before processing 62.5 238.0 32.0 8.1 27.4 9.5
(1) 80.0 140.9 30.6 7.3 22.4 7.1
(2) 80.1 169.5 21.1 7.3 20.3 7.0
(3) 80.6 127.0 27.6 5.1 20.2 5.7
(4) 88.0 157.3 20.7 7.7 15.5 7.9
Note: (1) natural dehydration plus blanching; (2) natural dehydration without blanching; (3) salting-out
dehydration plus blanching; (4) salting-out dehydration without blanching.

Table 7-9. Effect of steam blanching on the loss of minerals in spinach

g/100g
Mineral Loss (%)
Not steam blanching Steam blanching
K 7.9 3.0 56
Na 0.5 0.3 43
Ca 2.2 2.3 0
Mg 0.3 0.2 36
P 0.6 0.4 36
Nitrite 2.5 0.8 70

Table 7-10. Effect of processing method on the Cu content in potato


(mg/100g fresh weight)

Fluctuati Fluctuati
Processing method Cu Processing method Cu
on (%) on (%)
Material 0.21 0.00 Potato mesh 0.10 -52.38
Water boiling 0.10 -52.38 French frying potato crisp 0.27 +28.57
Baking 0.18 -14.29 Fast potato 0.17 -19.05
Potato chip 0.29 +37.20 Peeling potato 0.34 +61.90

The effect of processing method on Cu content in potato shown in Table 7-10. Cu content
is slightly increased in the frying potato crisp and peeling potato.
Minerals 245

4.3. Effect of Storage on Mineral Contents in Foods

Minerals in foods can migrate from packaging materials. The contents of some minerals
in canned liquid and solid foods are given in Table 7-11. The contents of Al, Sn and Fe in
solid foods are evidently increased because solid foods repeatedly collide with the packing
material.

Table 7-11. Contents (g/Kg) of trace elements in some vegetable cans

Vegetable Can State Al Sn Fe


Mung bean La L 0.10 5 2.8
S 0.7 10 4.8
Bean La L 0.07 5 9.8
S 0.15 10 26
Small green pea La L 0.04 10 10
S 0.55 20 12
Parsley heart La L 0.13 10 4.0
S 1.50 20 3.4
Sweetcorn La L 0.04 10 1.0
S 0.30 20 7.4
Mushroom P L 0.01 15 5.1
S 0.04 55 16
Note: a. La = Lacquered can; P = T in plate can; b. L = Liquid, S = Solid.

REFERENCES
[1] Capasso, C; Carginale, V. Solution structure of MT-nc, a Novel Metallothionein from
the Antarctic Fish Notothenia coriiceps. Structure, 2003, 11, 435-443.
[2] ESHA Research. The Food Processor Plus, ESHA Research, Salem, OR, 1992.
[3] Ji, L; Huang, J; Mo, T. Bioinorganic Chemistry an Introduction (Second), Guangdong:
Zhongshan University Press, 2001
[4] Kan, JQ. Food Chemistry. Beijing: China Agricultural University Press; 2008.
[5] Liu, Y. An idea on trace elements as nutrients of organism. Journal of Peking University
(Natural Science), 1986, (3): 121.
[6] Nagy, L; Szorcsik, A. Equilibrium and structural studies on metal complexes of
carbohydrates and their derivatives. Journal of Inorganic Biochemistry, 2002, 89, 1-12.
[7] Sigel, H; Sigel, A. Metal Ions in Biological Systems. 1st edition. New York: Marcel
Dekker, 1995.
[8] Tang Renhuan. On the Biological Element Spectrum in Organism. Acta Scientiarum
Naturalium Universitatis Pekinensis, 1996, 32, 790-803.
[9] U. S. Department of Agriculture (1976-1986). Composition of Foods. Agriculture
Handbook Nos.8-1 to 8-16. Human Nutrition Information Service, USDA, Hyattsville,
MD.
[10] Yang, P. Introduction of Bioinorganic Chemistry, Xi‘an: Xi‘an Jiaotong University
Press, 1991.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 8

FOOD FLAVORS

Xiaoxiong Zeng1 and Guaoqing Huang2


1
College of Food Science and Technology, Nanjing
Agriculture University, Nanjing, China
2
College of Food Science and Engineering, Qingdao
Agriculture University, Qingdao, China

ABSTRACT
The special tastes or aromas of foods are contributed by abundant small-molecular
weight compounds and these compounds significantly affect the acceptability of foods by
consumers. This chapter firstly concerns the major tastes and their perception by human
beings. Then, the major flavor compounds in various foods, including fruits, vegetables,
and meats are elucidated in detail. Besides, reactions that contribute to the formation of
flavors during food processing and storage are also presented in detail in this chapter.
Foods should not only meet the needs of consumers on nutrition, but also provide
desirable flavors so that consumers can enjoy the foods. Flavor is an important aspect that
determines food quality, and affects the intake and absorption of nutrients in foods.
Flavor is an overall integrated perception of all contributing senses (smell, taste, sight,
feeling, and sound) at the time of food consumption. The evaluation and preference to
flavors differ significantly among individuals, areas, and ethnics. Flavor together with
nutritional value and security determine consumers‘ acceptance of foods.
The flavor of food is contributed by multiple compounds in addition to
environmental factors. The development of modern analysis techniques, such as
chromatography and mass spectrometry, provides great convenience for further study of
food flavor. However, since food flavor is a physiological perception, neither qualitative
nor quantitative methods can accurately measure or describe a food flavor yet.
248 Xiaoxiong Zeng and Guaoqing Huang

TASTE COMPOUNDS IN FOODS


Taste and Taste Compounds

Food Taste
A taste is a feeling caused by the stimulation of food compounds on taste receptors in the
mouth. Taste buds are the most important taste receptors in the mouth, followed by free nerve
endings. Taste buds locate on the surface of the tongue and the back of the oral cavity. Each
taste bud is the cluster of approximately 30-50 taste cells and taste receptors are distributed on
the membranes of these cells. Each taste bud is connected to the oral cavity through a hole on
the top. When foods are ingested, the taste compounds get contact with the corresponding
receptors in the taste cells through the hole and consequently a taste is perceived. Free nerve
ending acts as a micro-receiver and can identify different chemical substances. It is scattered
in the entire oral cavity and surrounded by sacs.
Different parts of the tongue are sensitive to different tastes. Each of the primary tastes is
associated with a specific area in the tongue. The tip of the tongue is most sensitive to sweet
and salty tastes, while sour seems to register more strongly on the sides of the tongue. Far to
the rear of the tongue are most of the receptors for bitter tastes.
The sensitivity to a taste is often described by threshold, which is the lowest
concentration required for eliciting a sensation. Threshold values are usually determined
using individuals representative of the general populations, and each panelist indicates
whether or not the compound can be detected. The concentration range where at least half
(sometimes greater) of the panelists can detect the compound is designated as the flavor
threshold. Table 8-1 lists the threshold values of some taste compounds.
Three kinds of threshold value are proposed based on measurement methods. Absolute
threshold value, also called sensory threshold value, is the lowest level of a taste compound
that can be perceived. It is determined by tasting a series of diluted taste compound solutions.
Differential threshold value is the smallest change in taste compound concentration that
consumers can detect. Terminal threshold value is the minimum concentration of a taste
compound, whose further increase does not enhance the perception. In this chapter, all
thresholds value mentioned refer to the absolute threshold value, except special instructions.

Taste classification.

Different taste classifications have been proposed due to the differences in culture and
dietary habits. In Japan, the primary tastes include sweet, sour, bitter, salty and pungent. In
European and American countries, six primary food tastes are identified, including sweet,
sour, bitter, salty, pungent, and astringent. In India, up to eight tastes are recognized as
primary tastes and they are sweet, sour, bitter, salty, pungent, insipid, astringent and
abnormal. In China, the original standard specifies only five primary tastes, including sweet,
sour, bitter, salty and pungent, but the astringent and delicious tastes are added later. From the
physiological point of view, only the sweet, sour, bitter and salty tastes can be perceived by
taste receptors. The pungent taste is the pain feeling when pungent substances stimulate oral
mucosa, nasal mucosa, skin and trigeminal nerve. The astringent taste is the response of
tactile nerves to protein aggregation. Both the tastes affect the flavor of foods independently.
The presence of delicious compounds enhances the sensation of other tastes. In European and
American countries, the delicious taste is excluded from the primary tastes.
Food Flavors 249

Factors affecting taste perception.

The perception of tastes is affected by many factors. In addition to dietary habits, health
status, age and other individual factors, taste perception is also affected by the following:
Temperature: the optimum temperature for taste sensation is between 10-40 °C and taste
receptors are the most sensitive at 30 °C. When the temperature is lower than 10 °C or higher
than 50 °C, the sensation becomes dull. It can be seen from Table 8-1 that the salty taste is the
most sensitive to temperature variation and the sour taste of citric acid is the least sensitive.
Solubility: the intensity of a taste is related to the solubility of the taste compound,
because taste receptors or free nerve endings are stimulated only by dissolved compounds.

Table 8-1. Threshold values of several taste compounds

Threshold (%)
Taste compounds Taste
25 °C 0 °C
Quinine sulfate Bitter 1.0×10-4 3.0×10-4
Sucrose Sweet 0.1 0.4
Sodium chloride Salty 0.05 0.25
Citrate Sour 2.5×10-3 3.0×10-3

Generally, taste compounds that dissolve quickly are perceived in a very short time and
the sensation disappears very fast. For example, sucrose is readily soluble. Its sweetness is
perceived rapidly after ingestion, but the sensation lasts only a short time. In contrast, the
sweetness of saccharin is perceived slowly, but the sensation can last for a relative long time.
Presence of other taste compounds: Interactions might occur between different taste
compounds.

a. Enhancing: the presence of a taste compound could enhance the intensity of another
taste. For example, table salt in concentration 0.017% enhances the sweetness of
sucrose solution (15 %).
b. Modification: the presence of a taste compound might change the taste of another
compound. For example, water tastes sweet if table salt or quinine has been
perceived before.
c. Elimination: the presence of a taste compound reduces or eliminates the intensity of
another taste. For example, any one of sucrose, citric acid, table salt, and quinine can
reduce the taste intensity of the other three compounds. In wines or beverages, the
sweetness of sugars weakens the sour taste and vice versa.
d. Multiplication: when two taste compounds are present at the same time, the intensity
of the tastes increases significantly. For example, the mixture of sodium glutamate
and 5‘-inosinic acid has markedly improved delicious taste than alone and maltol
significantly enhances the sweet taste in beverages and candies.
e. Adaptation: adaptation refers to the gradual decline of taste intensity with prolonged
stimulation. The time required for the recovery of taste sensation varies with tastes.
The time needed for recovery of sour taste sensation after adaptation is 1.5~3 min
and that for sweetness, bitterness, and salty is 1~5 min, 1.5~2.5 min and 0.3-2 min
respectively.
250 Xiaoxiong Zeng and Guaoqing Huang

The interactions between tastes are very complicated and are affected by both
psychological and physicochemical factors. The mechanisms involved are not well
understood yet to present.

Sweet Taste
Sweetness is the most popular taste for consumers. It improves the palatability and
certain properties of foods. The intensity of sweetness is expressed in relative sweetness by
using 10 % sucrose solution as reference. Table 8-2 lists the relative sweetness of several
sweeteners.
The mechanism of sweet taste can be explained by the AH/B through proposed by Robert
Shallenberger and Terry Acree in 1963. It is supposed that the initial event in the perception
of sweetness is the hydrogen bonding of a stimulant to receptors on the tongue. Each sweet
compound possesses an electronegative atom A (usually N or O) covalently connected with a
hydrogen atom (AH). Hence, the AH can be hydroxyl (-OH), imino (-NH) or amino (-NH2)
group. AH functions as the proton donor. The sweet compound contains another
electronegative atom B (usually N, O, S, or Cl), which is 0.25-0.4 nm away from the AH
group and acts as the proton receptor. The sweetness receptor contains the AH/B unit. When
the AH/B unit of a sweet compound binds to the AH/B unit in the receptor through hydrogen
bonding, the taste nerve is stimulated and the sweet taste is perceived. Figure 8-1 shows the
AH/B structure of chloroform, saccharin and glucose.
This theory has been successfully used to explain the sweet sensation of many
compounds. However, this theory has some limitations. It can not explain why the sweet
intensities of sugars and D-amino acids that contain the AH/B structure differ significantly
and why the optical isomers of amino acids have different tastes.

Table 8-2. Relative sweetness of several sweeteners (with sucrose as 1.0)

Relative
Sweetener Sweetener Relative sweetness
sweetness
β-D-fructose 1.0-1.75 D-tryptophan 35
α-D-glucose 0.40-0.79 glycyrrhizic acid 200-250
α-D-galactose 0.27 saccharin 200-700
β-D-mannose 0.59 Naringin dihydrochalcone 100

Xylitol 0.9-1.4 Neohesperidin dihydrochalcone 1500-2000

Chloroform Saccharin Glucose

Figure 8-1. AH/B relationships of chloroform, saccharin, and glucose [1].


Food Flavors 251

Sweet receptor

Figure 8-2. Schematic showing the relationship between AH/B and γ sites in the saporous sweet unit for
β-D-fructopyranose [1].

To solve these problems, Kier extended the AH/B theory. He proposed that each sweet
compound also contains a lipophilic region γ in addition to the AH/B structure. The lipophilic
region might be methylene (-CH2-), methyl (-CH3) or phenyl (-C6H5) groups. All the three
active units (AH, B and region γ) must be arranged correctly in space so that the units can
contact with receptor molecules. The relationship between the three groups can be illustrated
in Figure 8-2.

Bitter Taste
The bitter taste is unpleasant on its own, but its combination with other tastes can provide
foods with special flavor, such as in tea, coffee, beer, and balsam pear. Strychnine is the
bitterest substance (threshold 0.0016 %) ever found and quinine is often used as reference in
the assessment of bitter intensity of other substance.
The sensation of the bitter taste is similar to that of the sweet taste. Bitter compounds
posses the AH/B entity and can interact with receptors through hydrogen bonding. The
difference is that the proton donor (AH) in bitter compounds are –OH, –C(OH)COCH3, –
CHCOOCH3, or –NH and the proton donor (B) is –CHO, –COOH, or –COOCH3. In addition,
the distance between AH and B is 0.15 nm, which is much less than that in sweet compounds.
Naturally occurring bitter compounds are alkaloids, terpenoids, and glycosides in plants
and bile in animals.
Caffeine, theobromine, theophylline: caffeine, theobromine and theophylline are
derivatives of purine and are important bitter substances in foods. Caffeine occurs in tea,
coffee and cacao, and theophylline is found in cacao and tea (Figure 8-3). All the three
alkaloids can stimulate the central nervous system.
Naringin and neohesperidin: naringin and neohesperidin are main bitter taste compounds
in citrus fruits and are mainly distributed in the peel of the fruits. Both the two compounds are
flavanone glycosides and are water soluble. The bitterness of naringin is related to the l→2
glycosidic bond between rhamnose and glucose. Enzymatic hydrolysis of the linkage yields
products without bitterness (Figure 8-4).
252 Xiaoxiong Zeng and Guaoqing Huang

R1 N N R3 R1=R2=R3=CH3 caffeine
R1=H R2=R3=CH3 theobromine
R1=R2= CH3 R3= H theophylline
O N N

R2

Figure 8-3. Structure of caffeine, theobromine and theophylline.

Enzyme

Figure 8-4. Debittering site of naringin by naringinase.

humulone cis-isohumulone trans-isohumulone

Figure 8-5. Thermal isomerization of humulone to isohumulone [2].

R3

COOH
R1=R2=OH R3=H chenocholic acid
R1=R3=OH R2=H deoxycholic acid
R1=R2=R3= OH cholic acid
R1 R2

Figure 8-6. Structures chenocholic acid, deoxycholic acid and cholic acid.

Bitter substance in beer: the bitter taste of beer is contributed by the bitter substances
originally contained in hops and those generated during brewing. The major bitter substances
in beer are α-acids, such as humulone, cohumulone, adhumulone. Hops are often added as the
mashed malted barely (or malt extract) is boiled in water. During this boiling process the
modestly bitter α-acids undergo thermal isomerization to form extremely bitter iso-α-acids.
(Figure 8-5).
Bile acid: bile acid is an extremely bitter fluid secreted by the liver of most vertebrates. It
is involved in the digestion of lipids in the small intestine and the adsorption of fat-soluble
vitamins. The major bitter compounds in bile acid are chenocholic acid, deoxycholic acid, and
cholic acid (Figure 8-6). If bile acid leaks from gallbladder during the processing of livestock,
poultry and aquatic products, the bitter taste cannot be removed even after repeated washing.
Food Flavors 253

Sour Taste
The sour taste is produced by hydrogen ion of organic acid, inorganic acids and acidic
salts. Moderate sour intensity yields pleasant sensation and enhances appetite. Generally, the
intensity of the sour taste is positively related to the concentration of hydrogen ion in
solutions. When the H+ concentration is too high (pH <3.0), the sour taste becomes
intolerable and sour intensity changes can no longer be perceived. The sensation of the sour
taste is affected by counter ion species and the buffering capacity of food medium. For
example, the sour intensities of five common acids in the same pH are in the following
decreasing order: acetate acid > formic acid > lactic acid > oxalate acid > hydrochloride yield.
Anions decide the characteristics of the sour taste, which is why the sour taste of sour
compounds differs.
Acidulants are important additives in food processing. The compounds not only impart
foods with desired sour taste, but also inhibit the growth of microorganisms. The most
commonly used acidulant in food industry is acetic acid, followed by citric acid, lactic acid,
tartaric acid, gluconic acid, malic acid, fumaric acid and phosphoric acid. Acetic acid is the
major active component of vinegar. Citric acid is the most widely used acidulant in food
industry. Malic acid is used to mask the lingering bitterness of synthetic sweeteners. Gluconic
acid-δ-lactone can produce gluconic acid upon heating and is hence a slow-acting acidulant. It
can be used as leavening agent in biscuit processing and curing agent in tofu preparation.
Phosphoric acid is mainly used in the production of cola-type beverages.

Salty Taste
The salty taste is contributed by neutral salts and it is an indispensable and the most
fundamental taste in foods. This taste is the compromise of the effects of dissociated anions
and cations. Cations produce salty taste, while anions suppress the sensation of salty taste and
elicit other undesirable tastes. Whether an inorganic salt is bitter or salty depends on the
diameters of cations and anions. If the sum of the diameters of cation and anion of a
compound is less than 0.65 nm, the salt tastes salty; otherwise, the salt tastes bitter. For
example, the sum of the diameters of Mg2+ and Cl- is 0.85 nm and MgCl2 tastes quite bitter.

L-sodium
L-sodium glutamate
glutamate (MSG) (MSG) Inosine-5‘-monophate
Inosine-5'- (5‘-IMP).
monophosphate (5′-IMP)

Sodium chloride is the only substance known to evoke a purely salty taste in any
concentration that is suprathreshold. The optimum concentration of NaCl in liquid foods is
0.8-1.2 %. Because excessive intake of NaCl can cause negative impacts on health, the
development of NaCl substitutes has attracted much attention. For example, a mixture of 20
% of KCl and 80 % NaCl has been developed as a low-sodium salt.
Delicious Flavor
The delicious flavor is a very complex sensation. The flavor enhances the appetite of
consumers and makes foods tasted more delicious. When the concentration of a delicious
254 Xiaoxiong Zeng and Guaoqing Huang

compound in foods is higher than threshold value, it evidently increases the delicious taste of
the food, and enhances the original flavor of the food even its concentration is lower than its
threshold value. Hence, delicious compounds are also known as flavor enhancers.
Delicious compounds can be divided into amino acid, nucleotide and organic acid types
and their typical representatives are L-sodium glutamate (MSG), 5‘-inosine monophosphate
(5‘-IMP), and sodium succinate respectively. MSG is the first discovered and commercially
available flavor enhancer. It occurs widely in nature and is abundant in kelps. 5‘-IMP is
widely found in chickens, fishes and broths and 5‘-IMP in animals is mainly generated by the
degradation of ATP in muscles. 5‘-Guanosine monophosphate (5‘-GMP) is the major
delicious component in mushrooms. Sodium succinate has been identified in poultry,
livestock and mollusks and the highest content is detected in shellfishes. It is also present in
low levels in fermented products, such as soy sauce, sauce, and yellow rice wine. Aspartic
acid and its sodium salt are the main delicious substances in bamboo shoots, though their
intensities are lower than that of MSG. The combination of IMP, GMP and MSG
significantly enhance the delicious taste of MSG. For instance, the delicious intensity of the
mixture containing 1 % IMP, 1 % GMP, and 98 % MSG is four times of that of MSG alone.

Pungency
Certain compounds found in some spices and vegetables cause characteristic hot, sharp
and stinging sensations that are known collectively as pungency. Pungent compounds
enhance the appetite and increase the secretion of digestive juice of consumers and they are
indispensable condiments in daily life. Based on sensation, natural pungent compounds are
divided into hot, aromatic and irritative types.
Hot compounds: pungent compounds of this type are odorless and can cause the burning
feeling in mouth. The pungent compounds in chili, black pepper and zanthoxylum belong to
this type.

Chili.

Capsaicinoids are the vanillyl amides of monocarboxylic acids with varying chain length
(C8~C11) and capsaicin is the active component in chili pepper. The contents of capsaicin in
chili peppers vary significantly with species. The contents of capsaicin in red chili, horn red
chili, Sam chili, and Uganda chili are 0.06 %, 0.2 %, 0.3 %, and 0.85 %, respectively.

Capsaicin

Black pepper.

Piperine is the major pungent compound in black pepper. Pepperine is an amide


compound and has three isomers, of which the isomers with more cis double bonds possess
Food Flavors 255

high pungency intensity. The liability of black pepper to light and storage is due to the
isomerization of piperine isomers.

Pipirine

Chinese prickly ash.

The main pungent compound in Chinese prickly ash is suberosin and it also an amide
compound.
Aromatic compounds: pungent compounds of this type are volatile and aromatic.

Ginger.

Zingiberols are the main active components responsible for the pungency of fresh ginger.
The carbon chain length on the lateral side of hydroxyl in the side chain on the ring is
different (n=5-9). When fresh ginger is dried, zingiberols are converted to more pungent
gingerols through dehydration. When ginger is heated, the side chain of zingiberols breaks to
produce zingerone, which is a moderate pungent compound.

Zingiberols Zingerols Zingerone

Cloves and nutmeg.

The main pungent components in the two species are eugenol and isoeugenol
respectively.

Eugenol Isoeugenol
Irritative compounds: in addition to tongue and oral mucosa, compounds of this type are
also irritative to nose and eyes.
256 Xiaoxiong Zeng and Guaoqing Huang

Mustard, radish, horseradish.

The pungency and irritability of these materials are contributed by mustard oil that is
generated through the hydrolysis of sinigrin. It is the collective of isothiocyanate. The
following active substances have been identified in mustard, radish, and horseradish:

CH2=CH CH2- NCS CH3CH=CH-NCS

allyl isothiocyanate propylene isothiocyanate.

CH3(CH2)3- NCS C6H5CH2-NCS

butyl isothiocyanate benzyl isothiocyanate.

Welsh onion, garlic, leek, and onion.

Disulfides are the major pungent and irritative substances in welsh onion, garlic, leek,
and onion. The pungent components of garlic are generated by the decomposition of alliin
and include diallyl disulfide and allyl propyl disulfide. The pungent components in leek and
onion are also sulfur-containing compounds. These substances are converted to sweet thiol
compounds when heated. This is why cooked onion and garlic taste sweet other than pungent.

Astringency
Astringency is the sensation of the aggregation of proteins in oral mucosa. Tannins and
polyphenols are major components in foods that cause astringency. Besides, some salts (such
as alum), aldehydes, organic acids (such as oxalic acid), and quinine acid are sometimes also
involved in astringency sensation. Mature fruits taste less astringent than immature fruits,
because polyphenols are decomposed, oxidized, or polymerized during maturation. Tea also
contains polyphenols, but their contents vary with processing methods. Red tea is less
astringent than green tea, because polyphenols are oxidized during fermentation. Astringency
is a characteristic taste of red wine. To obtain an acceptable astringency, measures must be
taken to reduce the contents of polyphenols in wines.

FLAVOR COMPONENTS IN FOODS


Aroma Compounds in Plant-Derived Foods

Fruits
The characteristic aromas of fruits are mainly contributed by esters, aldehydes and
terpenes, followed by alcohols, ethers and volatile acids. The contents of these compounds
increase gradually as fruits get mature. Table 8-3 lists the aromatic compounds in some fruits.

Vegetables
Generally speaking, the intensity of the aroma of vegetables is weaker than that of fruits.
However, some vegetable, such as onion, garlic, leek, and onion, possess unique and strong
aromas.
Food Flavors 257

Table 8-3. Main aroma components in some fruits

Fruit Major aroma components Minor aroma components


Apple Isoamyl acetate Volatile acids, ethanol,
acetaldehyde, geranium alcohol
Pear Isoamyl formate Volatile acids
Banana Isoamyl acetate, isoamyl isovalerate Hexanol, hexenal
Muskmelon Diethyl sebacate
Peach Ethyl acetate, agarolactone Volatile acids, acetaldehyde,
advanced aldehyde
Apricot Amyl butyrate
Grape Methyl o-aminobenzoate C4~C12 fatty acid esters, volatile
acids
Citrus Butyraldehyde, octyl aldehyde,
decanal, linalool
Peel Formic acid, acetaldehyde, alcohol,
acetone
Juice Phenylethyl alcohol, formic acid, ethyl
acetate

Fresh vegetables.

Many fresh vegetables have the smell of soil. This aroma is generated by methoxy alkyl
pyrazines, such as 2-methoxy-3-isopropyl pyrazine in fresh tomato and pee, 2-methoxy-3-
isobutyl pyrazine in green pepper, and 2-methoxy-3-sec-butyl pyrazine in red beet root. These
compounds are synthesized from the precursor leucine. The biosynthesis pathway of pyrazine
compounds in plants is shown in Figure 8-7.

enzyme

leucine

2-methoxy-3-isobutyl
pyrazine

Figure 8-7. Biosynthesis pathway of methoxy alkyl pyrazines in plant tissues [3].

Unsaturated fatty acids in vegetables can be oxidized by endogenous lipoxygenase to


produce peroxides, which are further decomposed to produce aromatic aldehydes, ketones
and alcohols.

Vegetables of the Liliaceae family.

The aroma of these vegetables is generated by sulfur compounds, such as dialkyl


thioethers, dialkyl disulfides, dialkyl trisulfides, and dialkyl tetrasulfides. In addition, thio-
258 Xiaoxiong Zeng and Guaoqing Huang

propanal, thiocyanate, thiocyanate esters, mercaptan, dimethylthiophene, and thiosulfinate are


also involved in aroma formation. These compounds are transformed by enzymes from their
precursors when plant tissues are damaged.
The aroma precursor of onion is S-(1-propenyl)-L-cysteine sulfoxide, which is converted
from cysteine. In aroma formation, the precursor undergoes rearrangement with the help of
allinase and generates 1-propenyl sulfenic acid and pyruvate. 1-Propenyl sulfenic acid is
unstable.

acetone alkyl
garlic alcohol ammonia

1-allyl-1-subsurfuracid

S-propionaldehyde-S-oxidate

Figure 8-8. Reactions involved in the formation of onion aroma [4].

Figure 8-9. Reactions involved in the formation of garlic flavor compounds [5].

Part of the compound is rearranged to lacrimatory sulfoxide thio-propanal and part is


converted to mercaptan, disulfide compounds, tri-sulfur compounds and thiophene (Figure 8-
8). All these compounds are involved in the characteristic aroma of onion.
2-Amino-3-[(S)-prop-2-enylsulfinyl]propanoic acid (alliin) is the aroma precursor of
garlic and it is degraded in a similar way to that of S-(1-propenyl)-L-cysteine sulfoxide in
onion (Figure 8-9). The generated 2-propene-1-sulfinothioic acid S-2-propenyl ester (allicin)
has a strong irritating smell. Allicin also undergoes rearrangement to produce mercaptan,
disulfide compounds and other aromatic compounds. Allicin together with diallyl disulfide
and methyl allyl disulfide produce the characteristic aroma of garlic.
The characteristic smell of chives flavor is contributed by dimethyl disulfide, dipropyl
disulfide and propyl propenyl disulfide and that of asparagus is generated by 1,2-disulfide-3-
cyclopentene and 3-hydroxy-butanone. 5-Methyl-2-hexyl-3-dihydro-furanone and
propylmercaptanare contribute to the characteristic smell of leek.

Vegetables of the Cruciferous family.


Cruciferous vegetables, such as mustard, radish, and horseradish, have strong pungent
smells. These smells are caused by isothiocyanate esters, such as 2-vinyl isothiocyanate, 3-
Food Flavors 259

propenyl isothiocyanate and 2-styryl isothiocyanate. Isothiocyanate esters are generated by


the enzymatic hydrolysis of glucosinolates. In addition to isothiocyanate esters, allyl
isothiocyanate (R-S-C=N) and nitrides are also formed during the hydrolysis (Figure 8-10).

Mushroom.

Figure 8-10. Reactions involved in the formation of Cruciferae flavors [6].

Figure 8-11. Reactions involved in the formation of lenthionine [7].

The aroma precursor of mushrooms is lenthionine acid. The precursor can be hydrolyzed
by S-alkyl-L-cysteine sulfoxide lyase to produce the aromatic lenthionine (Figure 8-11). In
addition, benzyl isothiocyanate, phenethyl isothiocyanate, and benzaldehyde cyanohydrin are
also involved in the aroma formation of mushroom.

Other vegetables.

The major aromatic compounds in cucumber are carbonyl substances and alcohols and its
characteristic aroma is contributed by 2-trans-6-cis-nonadienal, trans-2-nonene aldehyde and
2-trans-6-cis-nonadienol. Besides, 3-cis-hexenal, 2-trans-hexenal, and 2-trans-nonnenal also
affect the aroma of cucumber. These flavor compounds are synthesized from linoleic acid and
linolenic acid.
More than 80 kinds of volatile compounds have been identified in tomato, in which, 3-
cis-hexenal, 2-trans-hexenal, β-ionone, hexanal, β-damascenone, 1-penten-3-one, 3-methyl
butyraldehyde, are major active compounds for tomato aroma. In heated products such as
260 Xiaoxiong Zeng and Guaoqing Huang

ketchup, the aroma changes due to the formation of dimethyl sulfide, the increase of β-
ionone, β-damascenone and the decrease of 3-cis-hexenal and hexanal.
Potato contains only trace amounts of aromatic compounds. Pyrazines, including 2-
isopropyl-3-methoxy-pyrazine, 3-ethyl-2-methoxy-pyrazine and 2, 5-dimethoxy-pyrazine, are
the major active components in fresh potato. Volatile compounds in cooked potato are
carbonyl compounds (including saturated and unsaturated aldehydes, ketones and aromatic
aldehydes), alcohols (C3~C8 alcohols, linalool, neroli and geraniol), sulfur compounds
(mercaptan, sulfide and thiazole) and furan compounds.
A large variety of terpenes have been identified in the volatile oil of carrot, mainly
including γ-bisabolene, caryophyllene, and terpinolene. cis-γ-Bisabolene, trans-γ-bisabolene
and hypoxanthine contribute to the characteristic smell of carrot.

Aromatic Components in Tea


Aroma is an important factor that determines the quality of tea. The aroma type and
characteristic aroma compounds of teas are associated with tea varieties, growing conditions,
harvesting time, maturity and processing methods. Only tens of aroma compounds have been
identified in fresh tea leaves, but up to more than 500 aromatic compounds are found in
processed teas.

Aroma components in green tea.

Green tea does not undergo fermentation during processing and exhibits the typical
roasted and fresh smell. The first step of green tea processing is water removing and enzymes
are deactivated in this step. Hence, most aromatic components of green tea are original from
fresh leaves and only a few are formed during processing.
The main volatile components in fresh tea leaves are leaf alcohols (3-cis-hexenol, 2-cis-
hexenol) and leaf aldehydes (3-cis-hexenal and 2-cis-hexenal), which have a strong flavor of
grass. During processing, part low boiling point substances, such as leaf alcohols and leaf
aldehydes, are evaporated, while part leaf alcohols and leaf aldehydes are isomerized to trans-
leaf alcohols and trans-leaf aldehydes. These products have the faint scent and are the main
body of the aroma of green tea. The smells of high boiling point aromatic compounds, such as
linalool, benzyl alcohol, phenylethanol, and acetophenone, get exposed with the volatilization
of low boiling point substances. Linalool is an important active component and accounts for
10 % of total aromatic components in green tea. These high boiling point compounds have
pleasant smells and are important aromatic substances of green tea.
The characteristic aroma of teas harvested right before or after Ching Ming Festival is
contributed by disulfide ether and leaf alcohols and the aroma disappear gradually during
prolonged storage.

Semi-fermented tea.

Oolong tea is the representative of semi-fermented tea and its major aromatic components
include leaf alcohol, cis-jasmone, jasmine lactone, methyl jasmonate, nerolidol, benzyl
alcohol cyanohydrin, and ethyl acetate.
Food Flavors 261

CH2COOCH 3 HO CH CN

CH 2CH=CHCH 2CH3
OH
O O

O O

Cis-jasmone Jasmine lactone Methyl jasmonate Benzyl alcohol cyanohydrin


Nerolidol

Red tea

Red tea undergoes fermentation during processing and has strong aroma. During
processing, tremendous reactions occur to produce hundreds of aromatic components. Hence,
red tea has obviously different aroma from green tea. Alcohols, aldehydes, acids and esters
are major constituents of the aroma of red tea and violet ketone plays important role in the
formation of the characteristic aroma.

O O

O
O2 +
Cis-theaspirane β-carotene β-ionone β-damascenone

Figure 8-12. Oxidation and decomposition of β-carotene in red tea.

Carotenoids, amino acids, and unsaturated fatty acids are the aroma precursors for red
tea. During processing, β-carotene is oxidized and decomposed to ionone (Figure 8-12),
which is further oxidized dihydroaclinidiolide and theaspirone.
Unsaturated fatty acids in tea, especially linolenic acid and linoleic acid, undergo
enzymatic oxidation during processing to produce C6-C10 aldehydes and alcohols. Meanwhile,
the fatty acids are also esterified by alcohols to yield esters with different aromas, such as
benzyl acetate, ethyl phenylacetate, methyl benzoate, and methyl salicylate. These
compounds have important effects on the aroma of tea.
Amino acids are also decomposed by enzymes to produce aldehydes, alcohols, and acids.
These compounds also contribute to the aroma of red tea.

Aromatic Compounds in Animal-Derived Foods

Livestock and Poultry Meat


Raw pork contains more than 300 kinds of volatile compounds, of which, most are
hydrocarbons, aldehydes, ketones, alcohols, esters, furan compounds, nitrogenous compounds
and sulfur compounds. The aroma of different raw meats varies and depends mainly on lipid
composition. Raw beef or pork has no special odor, but raw mutton and dog meat possess
special smells. The goaty flavor of mutton is contributed by methyl fatty acids, such as 4-
methyl-capryli acid, 4-methyl-pelargonic acid and 4-methyl-capric acid, while the fishy smell
of dog meat is closely related to the presence of trimethylamine and lower fatty acids. The
special smell of sexually maturated male livestock is due to the secretion of gonad. For
example, two compounds, namely 5α-male-16-en-3-one and 5α-male-16-ene-3α-ol (Figure 8-
13), are responsible for the strong odor of the meat of emasculated male pig.
262 Xiaoxiong Zeng and Guaoqing Huang

enzyme

Pregnenolone 5α-male-16-en-3-one

Figure 8-13. Formation of characteristic aromatic components in boar [8].

Aromatic compounds in cooked meats are generated in three ways: lipid oxidation and
hydrolysis; Maillard reaction between amino acids or proteins and reducing sugars; and
further decomposition or recombination of flavor compounds. The aroma compositions of
cooked meats vary with the cooking temperature and processing methods. Cooked pork
contains reduced volatile compounds (mainly aldehydes, ketones, carboxylic acids and sulfur
compounds). Some non-volatile compounds, including free amino acids, peptides,
carbohydrates, vitamins and nucleotides, are important aroma precursors of livestock and
poultry meats. When heated, the precursors undergo various chemical reactions to yield
characteristic aromas.
When fat-containing beef is cooked, abundant volatile compounds are produced,
including fatty acids, aldehydes, ketones, alcohols, ethers, furan, pyrrole, lactones, aromatic
hydrocarbons, sulfur compounds (thiazole, thiophene, alkyl sulfur, sulfide, disulfide
compounds) and nitrogenous compounds (oxazole, pyrazine). To present, more than 600
volatile compounds have been identified in cooked beef, of which acidic compounds have
only minor effect on the aroma. Instead, thiophenes, furan, pyrazine compounds and pyridine
compounds contribute largely to the characteristic aroma of cooked beef. The composition of
the characteristic aroma of cooked pork is similar to that of beef, except that the contents of γ-
or δ-lactones converted from 4 (or 5)-hydroxyl fatty acid precursors are higher than in cooked
beef. Besides, cooked pork contains more unsaturated carbonyl and furan compounds than
cooked beef. Because mutton contains less free fatty acids and unsaturated fatty acids than
beef or pork, cooked mutton with less carbonyl compounds. The characteristic aroma of
cooked chicken meat is provided by sulfide and carbonyl compounds, in which, carbonyl
compounds, such as 2-trans-4-cis-decadien-1-al, are the most important active components.
The aroma of boiled meats is provided mainly by neutral compounds, such as sulfides,
furan-type and benzene-type compounds, while that of roasted meats is contributed mainly by
basic compounds, such as pyrazine, pyrrole, pyridine, and carbonyl compounds. However,
regardless of the processing method, sulfur compounds are the most important active
components for meat aroma. If sulfur compounds are removed, cooked meats will lose their
aroma. The content of hydrogen sulfide significantly affects the aroma of cooked meat. When
the content is too high, the odor of rotten egg is perceived; however, when the content is too
low, the aroma intensity is reduced. Smoked meats have unique aroma and tastes. The smoke
used contains phenols, formaldehyde, acetaldehyde, acetone, cresol, fatty acids, alcohols,
pyromucic aldehydes, and guaiacol, in which, fatty acids, phenols, and alcohols play
important role in the formation of the unique taste and aroma of smoked meats.
Lipids play an important role in the formation of the aroma of livestock meats. When
tallow is heated, it is decomposed and generates abundant compounds, including esters,
hydrocarbons, alcohols, carbonyl compounds, lactones, pyrazine and furan compounds, which
Food Flavors 263

are important active components of beef aroma. The same decomposition products have also
been detected in heated lard. When pork is cooked at temperatures lower than 100 °C, flavor
compounds derived from fat accounts for more than 50 % of total aroma compounds.

Aquatic Products

Volatile compounds in fresh aquatic products.

Generally, fresh marine fishes and freshwater fishes have only light odor and the odor is
mainly provided by volatile carbonyl compounds and alcohols, including aldehyde (C6, C8,
and C9), ketones and alcohols (such as 1-octene-3-one, 2-trans-nonyl aldehyde, cis-1-5-
octadiene-3-one and 1-octene-3-ol). These compounds are generated from the oxidation of
unsaturated fatty acids by lipoxygenase. As the freshness decreases, the compositions of the
odor change gradually and a characteristic fishy smell is perceived. This smell is caused by δ-
amino-valeraldehyde, δ-amino-valeric acid and hexahydropyridine compounds in fish skin
mucus, which are synthesized from basic amino acids. δ-Amino-valeraldehyde and δ-amino-
valeric acid have strong fishy smell. Because fish blood also contains δ-amino-valeraldehyde,
fish blood also smells fishy.

Volatile compounds in rotting fish.

Rotted aquatic products have disgusted odor. Ammonia, dimethylamine (DMA),


trimethylamine (TMA), methyl mercaptan, indole, skatole and fatty acid oxidation products
are major constituents of the odor. All these compounds are alkaline and can be neutralized
by acetic acid to eliminate the unpleasant odor.
In fresh fish, ammonia can also be generated in the formation of inosine monophosphate
(IMP) through the hydrolysis of adenine nucleotide (AMP) by AMP deaminase (Figure 8-14).
As the rot proceeds, free amino acids, urea and proteins are decomposed to produce large
amount of ammonia, such as the muscle of cartilaginous fishes contains high content of urea,
which can be decomposed by microbial urease to produce ammonia and carbon dioxide
(Figure 8-15). Hence, the fishes have strong smell of ammonia.
TMA is a main representative compound that causes the unpleasant odor of rotten fish
and its threshold is as low as 300-600 μg/kg. Fresh fishes contain trimethylamine oxide
(TMAO) instead of TMA. TMAO is odorless and is important in maintaining the osmotic
equilibrium between blood and tissues. It is found only in marine fishes. TMAO can be
reduced to TMA under the action of enzymes or microbes (Figure 8-16). TMA has been used
as an indicator of the degradation of unfrozen fishes.
DMA and formaldehyde are also the decomposition products of TMAO (Figure 8-16).
The odor intensity of DMA is lower than that of TMA.
264 Xiaoxiong Zeng and Guaoqing Huang

Figure 8-14. Generation of ammonia in AMP hydrolysis.

Figure 8-15. Formation of ammonia from urea.

Figure 8-16. Formation of volatile amines in marine fishes [9].

Volatile sulfur compounds are always detected in degraded marine fishes, such as
hydrogen sulfide, methyl mercaptan, dimethyl sulfide and diethyl sulfide. These compounds
are also involved in the unpleasant odor of marine fishes.
Marine fishes might exhibit the smell of oxidized fish oil or cod liver oil during storage
due to the oxidation of polyunsaturated fatty acids. Linolenic acid, arachidonic acid and
docosahexaenoic acid are the main unsaturated fatty acids in fish oil, their auto-oxidation
decomposition products can cause unpleasant odor. The smell of the oxidation products
depends on the degree of oxidation. In the early phase of oxidation, the products have the
smell of cucumber. As the oxidation proceeds, the smell of cod liver oil appears.
Food Flavors 265

FLAVOR COMPOUNDS FORMATION PATHWAYS


Lipoxygenase-Catalyzed Reactions

Lipid Oxidation
Lipoxygenase occurs widely in plants and catalyzes the oxidation of polyunsaturated
fatty acids. The produced peroxides are then decomposed by lyase to yield aldehydes,
ketones, alcohols and many other flavor compounds. Hexanal, synthesized by linoleic acid as
precursor (Figure 8-18.), is the flavor compound of apple, strawberry, pineapple, and bananas.
Lipid oxidation can also generate undesirable flavors. For example, the oxidation of linoleic
acid leads to the formation of the characteristic beany flavor of soybean. 2-trans-Hexenal and
2-trans-6-cis-nonadienol are the characteristic aroma compounds of tomato and cucumber,
and both the compounds are synthesized with linolenic acid as precursor (Figure 8-19).
Among the flavor compounds generated in the lipoxygenase-catalyzed pathway, C6
compounds has the fragrance of grass, C9 compounds generate aroma similar to that of
cucumber and watermelon, and C8 compounds possess the smell of mushroom or violet. C6
and C9 compounds are generally aldehydes and primary alcohols and C8 compounds are often
ketones and secondary alcohols.
The pleasant aroma of mature pear, peach, apricot and other fruits is generally
contributed by the β-oxidation products of long-chain fatty acid (C8-C12).

linoleic acid
lipoxygenase

peroxidelyase

hexanal

Figure 8-18. Formation of hexanal by the oxidation of linoleic acid.

+lipoxygenase
+aldehyde
++--yde lyase
lyase

oxygen acid

2-trans-hexenal 2-trans-6-cis-nonadienal aldehyde

Figure 8-19. Lipoxygenase catalyzed formation of aldehydes from long-chain polyunsaturated fatty
acids [10].
266 Xiaoxiong Zeng and Guaoqing Huang

Figure 8-20 β-oxidation of linoleic acid [11].

Figure 8-21. Pathways of aromatic compounds formation from leucine [11].

For example, 2-trans-4-cis-decadienoic acid ethyl ester is generated by the β-oxidation of


linoleic acid (Figure 8-20) and it is the characteristic aroma compound of pear. The β-
oxidation of lipids also generates C8-C12 hydroxyl acids. These compounds can be cyclized to
γ-lactones or δ-ketones by enzymes, of which, C8-C12 lactones have aroma similar to that of
coconut and peach.

Degradation of Branched-Chain Amino Acids


Branched-chain amino acids are important flavor precursors of mature fruits. The
characteristic branched-chain carboxylic acid esters formed in the post-ripening process of
banana, pear, kiwi fruit, and apples, such as isoamyl acetate and 3-methyl ethyl butyrate, are
derived from branched-chain amino acids (Figure 8-21).

Shikimic Acid Pathway


Shikimic acid is the precursor of the three essential aromatic acids. In addition to its
involvement in aromatic amino acids synthesis, it can also produce various volatile
compounds, as shown in Figure 8-22:
Food Flavors 267

enzyme eugenol

enzyme
enzyme p-cresol
cinnamyl

alcohol
lignin polymer

3-methoxy-4-hydroxybenzaldehyd(vanillin)

Figure 8-22. Volatile compounds produced in the synthesis of shikimic acid [12].

Figure 8-23. Structures of several important aromatic terpenoids.

In the synthetic pathway of shikimic acid, aromatic compounds (phenylalanine and other
aromatic amino acid) can be produced from intermediate product in the pathway. Besides
aromatic amino acid, the pathway can also form other volatile compounds related to essential
oil. Aromatic components which are formed by fumigation of food, some of them are also
formed from compounds in the shikimic acid pathway as precursors, such as vanillin.
Cinnamyl alcohol is an important aroma component in cinnamon perfume. Eugenol is the
main flavor and pungent ingredients in clove. Some important flavor compounds in shikimic
acid pathway are showed in Figure 8-22.

Terpenoids Synthesis
Terpenoids are important aromatic compounds of citrus fruits. Terpenoids containing two
or more isoprene units are nonvolatile and do not directly involve in aroma sensation.
Sesquiterpenes neral and ngcuka ketone are the characteristic aroma components of orange
and grapefruit respectively. Citral and limonene are monoterpenes and possesses the unique
smell of lemon and sour orange respectively. The enantiomers of terpenes may exhibit quite
different smells. For example, l-carvone [4(R)-(-) carvone] has a strong aroma of spearmint,
while d-carvone has the characteristic aroma of sweet wormwood (Figure 8-23).
268 Xiaoxiong Zeng and Guaoqing Huang

Heterolactic Fermentation Pathway


Microbial fermentation can yield abundant favor compounds and these compounds
significantly influence the taste and aroma of dietary products and alcoholic beverages.
Figure 8-24 shows the production of various flavor compounds in the heterolactic
fermentation of glucose and citric acid.
The products of microbial fermentation constitute the main body of the flavor of
alcoholic beverages. The flavor of beer is influenced by the presence of alcohols, esters,
aldehydes, ketones and sulfides, in which, isoamyl alcohol, α-phenyl ethanol, ethyl acetate,
isoamyl acetate, and ethyl benzene are the most important active components. Acetaldehyde,
diacetyl and hydrogen sulfide impart beer with the flavor of green apple and their contents
must be reduced to allowed ranges in post-fermentation. The flavor of Chinese white spirit is
affected by alcohols, esters, carbonyl compounds, phenols, and ethers. Aldehydes (mainly
aldehyde) are predominant aroma compounds in newly distilled wines and these components
make the wines tasted pungent. Furfural is usually not good for the flavor of wine, but it is an
important component which constitutes the sauce flavor of Maotai wine and its content
reaches up to 29.4 mg/L. Esters, especially the ethyl esters and iso-amyl esters of C2-C12 fatty
acid, ethyl phenylacetate, ethyl lactate, and phenethyl acetate, are determinants for the flavor
of Chinese white spirits.

Non-Enzymatic Reactions

Maillard Reaction
The Maillard reaction can yield a large number of aromatic compounds and their contents
and proportion vary with substrate type, heating duration, and temperature. When the reaction
occurs in a low temperature for a short time, aromatic l actones, pyran compounds and furan
compounds as well as Strecker aldehydes are produced. When the reaction proceeds in high
temperatures for a long time, pyrazine, pyrrole and pyridine compounds with baking aroma
are formed.
Pyrazine compounds are important flavor compounds in all bakery foods and thermally
processed foods. It is generally believed that pyrazine compounds are generated through the
Strecker degradation between amino acids and α-dicarbonyl compounds, which are the
intermediates of the Maillard reaction (Figure 8-25). Small sulfides formed in the Maillard
reaction also influenced the flavor of foods. For example, methionic aldehyde is the
characteristic aroma compound of boiled potatoes and cheese biscuits.
Methionic aldehyde is unstable and can be easily decomposed into methane thiol and
dimethyl disulfide, resulting in the increase of low molecular weight sulfides.
The thermal degradation produces H2S and NH3 are also involve in the flavor formation
through Maillard reaction. For example, H2S, NH3, and acetaldehyde are the thermal
degradation products of cysteine. They can react with hydroxy ketones formed in the Maillard
reaction to generate thiazoline, which has the flavor of cooked beef (Figure 8-26).
Food Flavors 269

Figure 8-24. Formation of volatile compounds in the heterolactic fermentation of citric acid and
glucose.

Figure 8-25. Formation of an alkyl pyrazine and small sulfur compounds through the Maillard reaction
[13].

cysteine

3-hydroxyl-2-butanone

Fig. 9-24. Methionine reacts with carbonyl compounds to form thiazoline

2,4,5-trimethyl-3-thiazoline

Figure 8-26. Formation of a thiazoline through the reaction between the thermal degradation products
of cysteine and carbonyl compounds [13].
270 Xiaoxiong Zeng and Guaoqing Huang

Thermal Degradation Reaction

Thermal decomposition of carbohydrates, proteins and fats.

Carbohydrates can be decomposed by heat in the absence of amines to produce a serious


of flavor compounds.
The thermal decomposition of monosaccharides and disaccharides produce mainly furan
compounds, accompanied by a small proportion of lactones, cyclodiones and other
substances. The compounds can be further decomposed to methylglyoxal, glyceraldehyde and
other low molecular weight volatile compounds.
Starch, cellulose and other polysaccharides are decomposed to furan compounds, furfural
compounds, maltol, as well as organic acids at temperatures below 400 °C.
The thermal degradation products of proteins and amino acids include hydrogen sulfide,
ammonia, pyrrole compounds, pyridine compounds, thiazole, thiophene, and many other
sulfur-containing compounds. Most of the products have strong smells. For the thermal
degradation of lipids, please refer to related contents in Chapter 4.

Degradation of vitamin.

When vitamin Bl is heated, it is decomposed and produces a large number of sulfur-


containing compounds, furan and thiophene and some of the products have meat flavor.
Ascorbic acid is unstable. When it is heated in the presence of oxygen, it is degraded to yield
furfural, glyoxal, glycerol aldehyde and other low-molecular aldehydes, of which furfural
compounds are important constituents of the aroma of cured tea, peanut and cooked beef.

Fat oxidation.

The non-enzymatic oxidation of lipids can cause rancidity. Nevertheless, moderate


oxidation of lipids imparts foods with desired flavor, such as in breads. Please refer to
Chapter 4 for the mechanism of lipid oxidation.

REFERENCES
[1] Shallenberer, RS; Acree, TE. Molecular theory of sweet taste. Nature, 1967, 216, 480-
482.
[2] DeTaeye, L; DeKeukeleire D; Siaeno E; Verzele M. Recent developments in hop
chemistry. In European Brewery Convention Proceedings. Amsterdam: European
Brewing Congress, 1977; 153-156.
[3] Morgan, ME; Libbey, LM; Scanlan, RA. Identity of the musty-potato aroma compound
in milk cultures of Pseudomonas taetrolens. Journal of Dairy Science, 1972, 55, 666
[4] Whitfield, FB; JH. Last Vegetables. In: Maarse, H. Volatile Compounds in Foods and
Beverages. New York: Marcel Dekker, 1991; 203-269.
[5] Shankaranarayana, ML; Raghaven, B; Abraham, KO; Natarajan, CP. Sulphur
compounds in flavours. In: Morton, ID; Macleod, AJ. Food Flavours, Part A,
Introduction. Amsterdam: Elsevier Scientific, 1982; 169-281.
Food Flavors 271

[6] Govindarajan, VS. Pungency: the stimuli and their evaluation. In: Boudreau, JC. Food
Taste Chemistry. Washington DC: American Chemical Society, 1979; 52-97.
[7] Hiraider, M; Miyazaki, Y; Shibata, Y. The smell and odorous components of dried
shiitake mushroom, Lentinula edodes I: relationship between sensory evaluations and
amounts of odorous components. Journal of Wood Science, 2004, 50, 358-364.
[8] Gower, DB; Hancock, M Bannister, LH. Biochemical studies on the boar pheromones,
5a-androst-16-en-3-one and 5a-androst-16-en-3a-ol, and their metabolism by olfactory
tissue. In: Cagan, RH; Kare, MR. Biochemistry of Taste and Olfaction. New York:
Academic Press, 1981; 7-31
[9] Hebard, CE; Flick, GJ; Martin. Occurrenceand significance of trimethylamine oxide and
its derivatives in fish and shellfish. In: Martin, RE; Flick, GJ; Ward, DR. Chemistry and
Bilchemistry of Marine Products. Westport: AVI Publishing, 1982; 149-304.
[10] Blank, I; Lin, J; Vera, FA; Weli, DH; Fay, LB. Identification of potent odorants formed
by autoxidation of arachidonic acid: structure elucidation and synthesis of (EZZ)-2,4,7-
tridecatrienal. Journal of Agricultural and Food Chemistry, 2001, 49, 2959-2965.
[11] Tressl, R; Holzer, D; Apetz, M. Biogenesis of volatiles in fruit and vegetables. In:
Maarse, H; Groenen, PJ. Aroma Research: Proceedings of the International Symposium
on Aroma Research, 1975 in Zeist, the Netherlands. Wageningen: Centre for
Agricultural Publishing and Documentation, 1975; 41-62.
[12] Wittkowski, R; Ruter, J; Drinda, H; Rafiei-Taghanaki, F. Formation of smoke flavor
compounds by thermal lignin degradation. In: Teranishi, R; Takeoka, GR; Guntert, M.
Flavor Precursors: Thermal and Enzymatic Conversions. Washington DC: American
Chemical Society, 1992; 232-243.
[13] Mussinan, CJ; Wilson, RA; Katz, I; Hruza, A; Vock, MH. Identification and some
flavor properties of some 3-oxazolines and 3-oxazolines and 3-thiazolines isolated from
cooked beef. In: Charalambous, G; Katz, I. Phenolic, Sulfur, and Nitrogen Compounds
in Food Flavors. Washington DC: American Chemical Society, 1976; 133-145.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 9

FOOD ADDITIVES

Linwei Liu1 and Shiyuan Dong2


1
Northwest AandF University, Yangling, Shaanxi, China
2
College of Food Science and Engineering, Ocean University of China,
Qingdao, China

ABSTRACT
Food additives are substances intentionally added by manufacturers to foods to
preserve flavor or enhance taste and appearance. A substantial amount of food additives
with different functions have been widely used in the food industry and these compounds
contribute largely to the development of the industry. Due to the increased concern on
food safety and the rapid development of analysis techniques, the definition of food
additive has evolved greatly. This chapter firstly describes the evolution of the definition
of food additives. Then, various food additives are introduced briefly according to their
classification and the functions, properties and related aspects of important additives are
described one by one.

1. INTRODUCTION
1.1. Definition

Food additives are regulated substances and therefore defined in law. The definition of
food additives was firstly proposed by the FAO/WHO Joint Expert Committee for Food
Additives (JECFA) in 1955 as ―non-nutritive substances added intentionally to food,
generally in small quantities, to improve its appearance, flavor, texture, or storage properties‖.
This definition covered a rather narrow range and did not include flavorings and nutrients.
Since then, the definition evolves gradually and various definitions have been proposed by
different countries or organizations.
According to the Chinese Hygienic Standards for the Use of Food Additives (GB2760-
2007), food additives refer to ―artificially chemosynthetic or natural substances to be added to
foods in order to improve food quality and color, flavor and taste, and meet the need of
274 Linwei Liu and Shiyuan Dong

preservation and processing technology. Nutritional fortification substances, flavoring agents


and processing aids, are also included in this definition as well‖.
The official definition of food additives in the European Economic Community (EEC) is
―any substance does not normally consumed as a food in itself and not normally used as a
characteristic ingredient of food, whether or not it has nutritive value, the intentional addition
of which to a food for a technological purpose in the manufacture, processing, preparation,
treatment, packaging, transport or storage of such food results, or may be reasonably expected
to result, in it or its by-products becoming directly or indirectly a component of such foods‖.
Federal Food Drug and Cosmetic Act, US (FFDCA) defines food additives as ―any
substance the intended use of which results or may reasonably be expected to result, directly
or indirectly, in its becoming a component or otherwise affecting the characteristics of any
food, including any substance intended for use in producing, manufacturing, packing,
processing, preparing, treating, packaging, transporting, or holding food, if such substance is
Generally Recognized As Safe (GRAS)‖.
Though the definitions vary, they specify three common functions for food additives.
Firstly, food additives improve food quality and meet consumers‘ requirements on flavor,
color, and taste. Secondly, food additives make food processing more reasonable, more
hygiene, more convenient and enhance the mechanized, automated, and scaled production of
foods. Thirdly, food additives contribute to saving resources, reducing cost, and providing
significantly social and economic benefits.

1.2. Classification

According to origin or source, food additives are divided into natural and artificial ones.
Food additives are also classified by functions in most cases. In China, food additives are
subdivided into 21 categories, including acidulant, anticaking agent, antifoaming agent,
antioxidant, bleaching agent, bulking agent, chewing gum base, coloring agent, color fixative,
emulsifier, enzyme preparation, flavor enhancer, flour treatment agent, coating agent, water
retention agent, nutrition enhancer, preservative, stabilizing and coagulating agent, sweeter,
thickener and others.
Among the 1500 kinds of food additives approved for use in China, more than 700 kinds
are spices and essence oils.

1.3. Regulation of Food Additives

To guarantee food safety, use of food additives is strictly regulated by national and
international laws. According to the Chinese Hygiene Standards for Use of Food Additives
(GB2760-2007), the use of food additives must comply with the following principles:
1. Food additives must be nontoxic, do not generate toxic compounds upon
decomposition and do not cause chronic poisoning symptoms after long-term intake of dose
allowed.
2. Food additives do not destroy the nutritional components of foods, reduce food quality,
or generate toxic compounds upon decomposition.
3. Food additives can not be applied for the purpose of adulteration or for shielding the
facts of food spoilage and food deterioration.
Food Additives 275

4. Food additives cannot be applied for the purpose of concealing the quality deficiency
caused by the food itself or by processing.
5. The application ranges, dosages, and residues of food additives must comply with
related national standards and regulations, and try to minimize the using amount to bring
about the desired result.
6. Processing aids must be removed after processing, except those with allowed residues.

2. ACIDULANTS
2.1. Functions

Acidulants or acidity regulators are some acids and their salts, which serve a variety of
functions shown as the following:

(1) Flavoring to provide a desired taste and serve to intensify, enhance, blend of modify
the overall flavor of the product.
(2) Reduction of the pH to prevent or retard the growth of microorganisms and the
germination of spores, and to increase the lethality of the process.
(3) Maintainer or establishment of pH by serving as buffering agents. Usually a
combination of free acids and salts are used.
(4) Chelation of metal ions (Cu, Fe) to assist in minimizing lipid oxidation, reducing
color changes and controlling texture in some fruits and vegetables.
(5) Alteration of the structure of foods including gels made from gums (pectin,
carrageenan), and proteins.
(6) Interaction with proteins and emulsifiers to modify the structure of foods such as
doughs, alter the heat stability of proteins, and to serve as an emulsifier in processed cheese.
(7) Modification of sugar crystallization in hard candy manufacturing.

2.2. Acidulants

The usual use of acidulants in food are acidic, phosphoric, citric, malic, succnic, tartaric,
lactic, gluconic, glycolic, fumaric and adipic acid. The pKa or pKa1 of them are 4.75, 2.1,
3.08, 3.4, 4.2, 3.2, 3.86, 3.60, 3.03 and 4.43, respectively. Their structures are shown in
Figure 9-1:
The differences between acidulates are flavor, acidity, metal chelating and antimicrobial
activity, solubility, hydroscopicity and cost. Citric, malic, tartaric and gluconic acids have
similar taste which compliant with making beverage and candy. But for cola drink, using
phosphoric acid is more suitable. Fumaric and acitic acid have good antimicrobial activity,
and acetic acid (vinegar) is an important condiment and functional agent for Chinese food,
mayonnaise and pickles. Tartaric acid is often used to make leavening (bulking agent) owing
to its lower solubility.
276 Linwei Liu and Shiyuan Dong

Figure 9-1. Sketch of structures of some acidulants added in food.

Gluconolactone can be used to make Doufu and also used as a ingredient for making the
leavening because it can change to gluconic acid gradually in hot water.
Lactic acid has special taste, which compliant with many dairy products and fermented
food, such as yoghourt and alcohol. Phosphoric, citric, malice, succinct and tartaric acids are
metal chelating agents, they can be used as antioxidant synergist.

3. SWEETENERS
3.1. Type and Functions

There are two types of sweeteners: caloric (nutritive) and noncaloric (non-nutritive). The
caloric sweeteners provide about 4 calories per gram. The noncaloric varieties provide zero
calories.
Caloric sweeteners provide sweet flavor and bulk when added to food. They also
maintain freshness and contribute to product quality. Caloric sweeteners act as a preservative
in jams and jellies, and a flavor enhancer in processed meat. They provide fermentation for
breads and pickles, bulk to ice cream, and body to carbonated beverage. Some caloric
sweeteners are rich in natural resources, e.g., sucrose and fructose. Some are mainly made by
processing sugar or other small molecule compounds, e.g., molasses and sugar alcohols.
Sugar alcohols are low glycemic sweetener that is safe for diabetics, and helps manage
healthy glucose levels, and does not promote tooth decay.
Non-caloric sweeteners are used in place of caloric sweeteners in some foods. They do
not provide calories, but they do provide the sweet taste. Most non-caloric sweeteners are
chemically sythenic. Some of them, e.g., saccharin, acesulfame K and sucralose are more
stable than sugar to high temperature and other conditions of food processing, e.g., they do
not go in for non enzyme browning. Almost all of them can use in the diet for diabetics and
obesity patients.
Food Additives 277

3.2. High Fructose Corn Syrup (Hfcs)

This sweet enhancer is derived from cornstarch and is a mixture of 55% fructose and
45% sucrose. Starches are treated with enzymes that convert glucose to fructose, which
results in a sweeter product. The sweeters are used in many mass-produced foods, including
soft drinks, baked goods, jelly, syrups, condiments (like ketchup), fruits and desserts.

3.3. Sugar Alcohols

Sugar alcohols belong to polyols family which has many functions in food, such as
increasing viscosity, modifying sugar crystallization, being water binder, plasticizer,
cryoprotectant, and non-cariogenic sweetener. In China and many other countries, xylitol,
maltitol, sorbitol and mannitol are allowed to add into food.
Xylitol is odorless and a white crystalline powder with a pleasant and sweet taste, and its
energy value is 16.72 kJ/g. It has the same sweetness and bulk as sucrose with one-third fewer
calories and no unpleasant aftertaste. It quickly dissolves, produces a cooling sensation in the
mouth and reduces the development of cavities. Xylitol occurs naturally in many fruits and
vegetables and is even produced by the human body during normal metabolism. It is
produced commercially from plants such as birch and other hard wood trees and fibrous
vegetation, or from xylose hydrogenation. It can be used into any food with the amount as the
required of food making because its safety level is high.
Maltitol (4-O-α-glucopyranosyl-D-sorbitol) has 75-90% of the sweetness of sucrose and
its food energy value is 8.8 kJ/g. It is used to replace table sugar because it has fewer calories,
and does not promote tooth decay and has a somewhat lesser effect on blood glucose. Maltitol
is made by hydrogenation of maltose obtained from starch. It is especially used in production
of sweets: sugarless hard candies, chewing gum, chocolates, baked goods, and ice cream. Its
similarity to sucrose allows it to be used in syrups with the advantage that crystallization is
less likely. Maltitol may also be used as a plasticiser in gelatine capsulesand as a
humectant.http://en.wikipedia.org/wiki/Maltitol - cite_note-1#cite_note-1 Maltitol does not
brown and caramelize after liquifying by exposing to intense heat. It is somewhat more
slowly absorbed than sucrose which makes it somewhat more suitable for people with
diabetes than sucrose. It can use with quantity stipulated by use standard for different food
categories.
Sorbitol, also known as glucitol, is a sugar alcohol that the human body metabolises
slowly. It is obtained by the reduction of glucose changing the aldehyde group to an
additional hydroxyl group. It also occurs naturally in many stone fruits and berries from trees
of the genus sorbus. It provides dietary energy 2.6 kJ per gram. and is often used in diet foods
including diet drinks and ice cream, mints, cough syrup, and sugar-free chewing gum.
Sorbital sometimes is used as a sweetener and humectant in cookies. Ingesting large amounts
of sorbitol can lead to abdominal pain, gas, and mild to severe diarrhea. In China, the
maximum level of use to each food is 5.0 g/kg.
Mannitol is also used as a sweetener for diabete patient. Since mannitol has a positive
heat of solution, it is used as a sweetener in "breath-freshening" candies, the cooling effect
contributing to the fresh feel. The pleasant taste and mouth feel of mannitol also make it as a
popular excipient for chewable tablets. It can be used a little in food coating for flavor.
278 Linwei Liu and Shiyuan Dong

3.4. ISO-Maltulose

Iso-maltulose, also known by the trade name Palatinose, is a disaccharide that is


commercially manufactured from sucrose hydrolysed via bacterial fermentation. It is a natural
constituent of honey and sugar cane and has a very natural sweet taste. It is particularly
suitable as a non-cariogenic sucrose replacement. Iso-maltulose is fully absorbed in the small
intestine as glucose and fructose. Like sucrose, it is fully digested and provides the same
caloric value of approximately 4 kcal/g. However, it is low-glycemic and low-insulinemia.
Because isomaltulose is released to the blood slowly, this sweetener avoids the sudden
increase of drop of blood glucose level. This leads to a more balanced and prolonged energy
supply in the form of glucose. In China, iso-maltulose is allowed to use in many foods and the
quantity of use is decided by the needing of food manufacture.

3.5. Noncaloric Sweeteners

There are several noncaloric sweeteners, such as saccharin, aspartame, acesulfame K, and
sucralose and their chemical structures are shown in Figure 9-2:
(1) Saccharinhttp://en.wikipedia.org/wiki/Saccharin - cite_note-1#cite_note-1 is a
colorless, crystal and artificial sweetener. The basic substance, benzoic sulfimide, has
effectively no calories and is many times sweeter than sucrose (about 300-700 times as
sucrose) [1], but has an unpleasant bitter or metallic aftertaste, especially at high
concentrations. In countries where saccharin is allowed as a FD, it is used to sweeten products
such as drinks, candies, medicines, and toothpaste. In China, the maxium usage quantity of
saccharin for food is 0.15 g/kg. Many studies have been performed on saccharin, some
showing a correlation between saccharin consumption and increased frequency of cancer in
rats (especially bladder cancer) and others finding no such correlation. No study has ever
shown a clear causal relationship between saccharin consumption and health risks in humans
at normal doses, though some studies have shown a correlation between consumption and
cancer incidence.
(2) Aspartame is 200 times sweeter than sugar in typical concentrations. Aspartame has a
caloric value of 17 kJ per gram, so the minimum quantity of aspartame which needs to
produce a sweet tast is so small that its caloric contribution is negligible. Therefore, it is a
popular sweetener for those trying to avoid calories from sugar. The taste of aspartame is not
identical to that of sugar. Blends of aspartame with acesulfame K taste more like sugar, and to
be sweeter than either substitute used alone.
Like many other peptides, aspartame could be hydrolyzed into its constituent amino acids
under conditions of high temperature or pH. This makes aspartame undesirable as a baking
sweetener.. At room temperature, it is the most stable at pH 4.3, so that it is reasonably stable
in soft-drinks. In the powdered beverages, the amine in aspartame can undergo Maillard
reaction with the aldehyde groups present in certain aroma compound. The safety of
aspartame has been studied thoroughly, and the scientific evidence indicates it is safe at
current level of consumption as a non-nutritive sweetener.
(3) Acesulfame K, also known as cesulfame potassium, is a white crystalline powder. It is
180-200 times sweeter than sucrose with a slightly bitter aftertaste, especially at high
concentrations.
Food Additives 279

Figure 9-2. Structures of some noncaloric sweeteners.

Acesulfame K is often blended with other sweeteners (usually sucralose or aspartame)


which give a more sugar-like taste, and each of them masks the other aftertaste. It is stable
under heat even under moderately acid or basic conditions, allowing it to be used in baking,
and it has been also used in beverages, tabletop sweeteners, desserts, puddings, baked goods,
soft drinks, dairy products, candies and canned foods to replace part sucrose. Research shows
that acesulfame K is safe to consume if the concentration in any edible form is less than 3%.
(4) Sucralose is approximately 600 times as sweet as sucrose (table sugar), twice as sweet
as saccharin, and 3.3 times as sweet as aspartame. Unlike aspartame, it is stable under heat
and over a broad range of pH conditions. Therefore, it can be used in baking or products that
require a longer shelf life. The commercial success of sucralose-based products stem from its
favorable comparison to other low-calorie sweeteners in terms of taste, stability, and safety.
In China, the maxium usage quantity of sucralose for many foods is 0.25 g/kg or less.
(5)There are other noncaloric sweeteners being used in different countries. Sodium
cyclamate is a water soluble chemical powder. It is 30 times as sweet as sucrose and stable in
most conditions of food processing. It is allowed to use in cold drink and pickle with the ML
equal to 0.65 g/kg. Alitame, made from amino acids, is an artificial sweetener about 2000
times sweeter than sucrose and has no aftertaste. It is relatively stable. Alitame was approved
for use in Mexico, Australia, New Zealand and China. Leaves of Stevia rebaudiana are a
source of several sweet glycosides of steviol. The major glycoside, stevioside, diterpenoid
glycoside are used in oriental countries as food sweetener. Its medical use is also reported as a
heart tonic. Besides, it is used against obesity, hypertension, and stomach burn and to lower
uric acid levels.

4. PRESERVATIVES
4.1. Definition

Preservatives or antimicrobial agents play an important role in today's supply of safe and
stable foods. Preservative means any substance which is capable of inhibiting, retarding or
arresting the process of fermentation, retarding acidification or other deterioration of food or
masking any of the evidence of putrefaction. Preservatives work by preventing the growth of
280 Linwei Liu and Shiyuan Dong

microbes (fungi, bacteria) or by killing the microbes. Food preservatives, low toxicity, are
commonly used in food processing. They should not pose significant health effects on
consumers during normal consumption.

4.2. Use and Limitation

To prolong the shelf-life of food against deterioration caused by microorganisms, the


preservatives are indispensable in the food industry. In several decades, increasing demand
for convenient foods and reasonably long shelf life of processed foods makes the use of
chemical food preservatives imperative because they are more effective and cheaper than bio-
preservative.
Traditional preservatives, such as wood smoke or vinegar, have been used for centuries.
However, they are not always safe. Smoking may introduce carcinogenic materials into the
food. Some of the commonly used preservatives, belong to inorganic chemicals, such as
sulfites, nitrate, and salt have also been used for centuries in processed meats and wine. The
use quantity of them are limited strictly to avoid their harmful affects on human health. Many
bio-preservatives are produced by microorganism; a few others are made from plant. They
also have some unhealthy effects on people.
Since there are not any preservatives being using in food without any danger, some
countries have set up their mandatory use standard of food preservatives. The using levels
must be obeyed the items of the standards to minimize risk.

4.3. The Choice of Preservative

The choice of antimicrobial agent has to be based on the antimicrobial spectrum of the
preservative, the chemical and physical properties of both food and preservative, the
conditions of storage and handling, and the assurance of a high initial quality of the food to be
preserved.
Many preservatives have broad bacteriostasis spectrum, but some others can only affect
on some species of microorganism. Salts of acids, such as sodium acetate, calcium citrate,
sodium benzoate, calcium propionate, and sodium nitrite all have antimicrobial properties.
Sulfur dioxide can be derived from a various sulfur additives such as bisulfite, and produce a
multitude of effects in food. Most preservatives only have antimicrobial effects at certain
range of pH because their states can be changed with the change of pH. Some preservatives
are unstable in high temperature, and some are dissoluble in water or oil. Therefore, the user
must know those properties before using them. Table 9-1 shows properties of some common
preservatives.

4.4. Preservatives in Common Use

4.4.1. Benzoic Acid


Benzoic acid naturally occurs in many types of berries, plums, prunes, and some spices.
As an additive, it is used as benzoic acid or as benzoate.
Food Additives 281

Table 9-1. Properties of some preservatives [2,3]

Compound Effective Effective Solubility Some food


against pH range applications
Benzoic acid Yeast, mold, 2.5-4.0 Water easily, Jam,juice,sauce
(benzoate) bacteria Oil slightly
Sorbic acid Yeast, mold, 3.0-6.5 Water easily, Meat, juice,
(sorbate) bacteria, Oil slightly bread
Parabens Bacteria,mold, 3.0-9.0 Ethnol easily, Water Sauce, beverage,
yeast slightly baking product
Dehydroacetic Mold,yeast, 3-10 Ethnol easily, Water Pickles,
acid bacteria slightly orange pulp
Sodium Mold 6-8 Water easily, Baking product,
propionate Ethnol easily, meat
Sulfites Bacteria,mold, 2.5-5.0 Water easily Wine,
yeast dried fruits
Nitrites Bacteria,mold, 4.0-5.5 Water easily Meat
yeast

The latter is used more often because benzoic acid is sparsely soluble in water, and
sodium benzoate is more soluble. The undissociated form on benzoic acid is the most
effective antimicrobial agent. The optimum pH range ranges from 2.5 to 4.0, which makes it
an effective antimicrobial in highly acidic foods. Benzoic acid and sodium benzoate are
widely used in fruit drinks, cider, carbonated beverages, sauce, jams and pickles with 1.0 g/kg
or less as the ML for different foods.

4.4.2. Sorbic Acid and Potassium Sorbate


Sorbic acid is a straight-chain, trans-trans unsaturated fatty acid, 2, 4-hexadienoic acid,
and it has a low solubility in water at room temperature. The salts of sorbic acid, such as
sodium, or potassium are more soluble in water. Sorbates are stable in the dry form; they are
unstable in aqueous solutions because they decompose through oxidation. The rate of
oxidation increases at low pH, high temp, light exposure. Sorbic acid and other sorbates are
so effective against yeasts and molds that they can be used in many foods, such as wine, fruit
juice, dried fruit, cottage cheese, meat, and fish products. Sorbates are most effective in
products with low pH including salad dressings, tomato products, carbonated beverages. In
China, its ML in most foods is 1.0 g/kg.

4.4.3. Propionic Acid, Sodium Propionate and Calcium Propionate


Propionic acid and its salt are effective against molds, but no effect on yeast. These
additives are mainly used in baked foods. The calcium salt is the most popular form used. In
China, its ML in many foods is 2.5 g/kg [4].

4.4.4. Sulfites
Sulfur dioxide and sulfites have been used as preservatives serving both as antimicrobial
substance and antioxidant. Instead of sulfur dioxide solutions, a number of sulfites, such as
sodium bisulfite, sodium metabisulfite, sodium sulfite, potassium bisulfite, sodium hydrogen
sulfite, sodium hyposulfite and potassium metabisulfite can be used. When dissolved in
water, they all yield active SO2 (see the following equations)[3]. The most widely used of
these sulfites is potassium metabisulfite.
282 Linwei Liu and Shiyuan Dong

SO2(gas) ↔ SO2(aq)
SO2(aq) ↔ H2O + H2SO3
H2SO3 ↔ H+ + HSO3-
HSO3- ↔ H+ + SO32-
2HSO3- ↔ S2O52- + H2O

All of these forms of sulfur are known as free sulfur dioxide. Meanwhile, the bisulfite ion
(HSO3-) can react with aldehydes, dextrins, pectic substances, proteins, ketones, and certain
sugars to form addition compounds, which are known as bound sulfur dioxide.
Sulfurous acid inhibits molds and bacteria to a lesser extent yeasts. For this reason, SO2
can be used to control undesirable bacteria and wild yeasts in fermentations without affecting
the SO2- tolerant cultured yeasts. The antiseptic activity of SO2 highly depend on the pH. The
lower the pH, the greater the antiseptic action of SO2. The amount of SO2 added to foods is
self-limiting because the product may develop an unpleasant off-flavor at concentration from
200 to 500 ppm. The use of SO2 is not permitted in foods that contain significant quantities of
thiamine, because this vitamin is destroyed by SO2.
Because SO2 is volatile and easily lost to the atmosphere, the residual levels may be
much lower than the amounts originally applied. In China, the ML of sulfites are calculated
by means of the residual levels in the finished food. Different food has different maxium
residual quantity. For example, the maximum residual quantities for fresh and dried fruits
processed are 0.05 g/kg and 0.2 g/kg, respectively.

4.4.5. Natamycin
Natamycin (Figure 9-3) is a naturally occurring antifungal agent produced by
Streptomyces natalensis. Natamycin has a very low solubility in water; however, it is
effective at very low levels. Natamycin has been used for decades in the food industry as a
hurdle to fungal outgrowth in dairy products, meats, and other foods. It has a neutral flavor
impact, and less dependence on pH for efficacy. It may be applied by spraying a liquid
suspension, by dipping the product in an aqueous suspension, or by mixing it into the product
in a powdered form along with cellulose on whole, shredded, or soft cheeses.

4.4.6. Nisin
Nisin is an antimicrobial polypeptide, with a molecular weight of 3500, produced by
some strains of Lactococcus lactis. Nisin-like substances are wide products from lactic acid
bacteria.

http://upload.wikimedia.org/wikipedia/commons/a/a3/Natamycin.svg

Figure 9-3. Structure of natamycin [5].


Food Additives 283

These inhibitory substances are known as becteriocins. Nisin can be used as a processing
aid against gram-positive organisms. It has been effectively used in preservation of processed
cheese. It is also used in the heat treatment of nonacid foods and in extending the shelf life of
sterilized milk [6].

5. ANTIOXIDANTS
5.1. Definition and Classification

Food antioxidants in the broadest sense are all of the substances that have some effects on
preventing or retarding oxidative deterioration that leads to rancidity, loss of flavor, color and
nutritive value of foodstuffs in foods.
Antioxidants can be classified into a number of groups: (1) Primary antioxidants which
can terminate free radical chains reactions and function as electron donors including the
phnolic antioxidants, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),
tertiary butyl hydroquinone (TBHQ), propylgallate (PG), natural and synthetic tocopherols
and so on. (2) Oxygen scavengers which can remove oxygen in a closed food system. Most
widely used compounds are Vitamin C, and related substances, ascorbyl palmitate, and
erythorbic acid (the D-isomer of ascorbic acid). (3) Chelating agents or sequestrants. They
remove metallic ions, especially copper and iron, that are powerful pro-oxidants. Citric acid is
widely used for this purpose. Amino acids and ethylene diamine tetraacetic acid (EDTA) are
examples of chelating agents. (4) Enzymatic antioxidants which can remove head space
oxygen dissolved, such as glucose oxidase. Superoxide dismutase can be used to highly
remove oxidative compounds from food systems. (5) Natural antioxidants which are present
in many spices and herbs. Rosemary and sage are the most potent antioxidant spices.

5.2. Mechanism of Antioxidation

Primary antioxidants are mostly phenolic compounds. They can react with free radicals,
which scavenge the active radicals and change themselves into lower active radicals. The
lower active radicals can react each other or with other radicals until becoming stable
molecules. That is why they can terminate free radical chains reaction, which is the main
oxidation process of lipids. Phenolic compounds are a mixture and in combination with a
chelating agent because different antioxidants have the synergistic effect. This kind
antioxidant should be added into food before serious oxidation has occurred and used with
small quantity. Otherwise, it may act as a pro-oxidant. For example, phenolic radicals can
promote ROOH decomposition to produce ROOO.

ROO AH 2 ROOH AH
ROO AH ROOH A
AH AH A AH 2

Chelating agents which have ability to bind metal ions have contributed significantly to
stabilization of food color, aroma and texture because traces of heavy metal ions can act as
284 Linwei Liu and Shiyuan Dong

catalysts for lipid oxidation. Many natural constituents of food can act as chelating agents, e.
g., carboxylic acids (oxalic, succinic), hydroxy acids (lactic, malic, tartaric, citric),
polyphosphoric acids (ATP, pyrophosphates), amino acids, peptides, proteins and porphyrins.
Both ascorbic acid and glucose plus glucose oxidase and catalase can deplete O2 in food
system. When they react with oxygen, ascorbic acid becomes to dehydroascorbic acid, and
glucose becomes to gluconic acid, and H2O2, another product of the oxidation of glucose, can
be broken down to water by catalase. When the oxygen has been depleted completely, the
food oxidation will stop. However, owing to VitC, oxidation is based on a complex
mechanism with some active substance giving birth to the midway of the reaction, adding less
quantity of VitC will prooxidant.

5.3. Common Antioxidants

The common used antioxidants in food are tocopherols, ascorbic acid, ascorbyl palmitate,
sodium isoascorbate, tert-butylhydroxyanisole (BHA), di-tert-butylhydroxytoluene (BHT),
tertiary butyl hydroquinone (TBHQ), propylgallate (PG), tea polyphenols, antioxidant of
glycyrrhiza, and so on. The structures of some of the antioxidants are shown in Figure 9-4.
Tocopherols and ascorbic acid are two vitamins, and their use in food have no danger but
expensive. Tocopherols and ascorbic acid, which are supplyed either natural or synthetic
equivalents, are less restricted in food using. In China, they can be used in food with any
quantities as the needs for food process. Ascorbyl palmitate, a derivative of ascorbic acid, has
good solubility in oil, so as to be a wide use as antioxidant which contains abundant oil or fat.
Sodium isoascorbate has almost the same function as ascorbic acid except it is not a vitamin
and cheaper than ascorbic acid. The use standard of FDs of China permit those antioxidant to
be added into many food with ML being 0.2-0.5 g/kg.
BHA, BHT and PG have been used, with powerful antioxidant function, for many years
in most countries. The first two are typical lipophilic artificial antioxidants and the last one
has some polarity. The first two are very stable during food process, but PG is unstable at
high temperature and discolour in presence of iron and metal complexes. The use of these
synthetic antioxidants is very restricted. The ML of them in most food categories is 0.1 g/kg
according to hygienic standards for uses of FDs in China.
TBHQ is more stable and less volatile than BHA at high temperature. TBHQ gives
excellent antioxidant potency to oils and fats.

Figure 9-4. Structure of some commercial antioxidants.


Food Additives 285

It can maintain freshness and quality of crude oils during long distance transportation,
and can offer protection to fried foods, thus enhancing their storage life and freshness. The
highest level of TBHQ permitted in the GSFA (USA) is 1 g/kg for frozen fish, fish fillet, and
fish products, and in the FDs use standard (China) is 0.2 g/kg for edible oils and fats, fried
food, curing meat class, and dried fishes. It is often used in conjunction with BHA, BHT and
PG to provide a synergistic antioxidant effect.
Tea polyphenols, antioxidant of glycyrrhiza and antioxidant of bamboo leaves are natural
antioxidants being extracted from tea, liquorice and bamboo. Tea polyphenols are water
soluble. Antioxidant of glycyrrhiza is unsoluble in water but in ethanol and ethyl acetate. The
antioxidant of bamboo leaves is soluble in ethanol. They are phenols substance, such as
catechins, flavonoids and phenolic acids. The ML of TP, AG and ABL in the FDs use
standard (China) are 0.3, 0.2 and 0.5 g/kg, respectively.
Rosemary extract is another natural antioxidant. The active ingredients are ursolic acid,
rosmarinic acid, conorsic acid, and conorsol. The ML of it in the FDs use standard (China) is
0.3 g/kg.
EDTA is abbreviation of disodium ethylene-diamine-tetraacetate, a famous chelating
agent in analysis chemistry. Phytic acid (inositol hexaphosphoric acid), orthophosphoric acid,
pyrophosphoric acid, triphosphoric acid, hexameta-phosphoric acid, tartaric acid, citric acid,
gluconic acid, and their salts are also good chelating agents. All of those chelating agent have
certain ability of antioxidant. In the use standard of FDs of China, the ML of EDTA and
phytic acid are 0.07 and 0.2 g/kg repectivesly. The orthophosphoric acid, pyrophosphoric
acid, triphosphoric acid, hexameta-phosphoric acid are classified also as water retention
reagents which can be use with high levels as 3-5 g/kg. The other chelating agents meationed
above can be used in food without level limits.

6. EMULSIFIERS AND STABILIZERS


6.1. General Description

The purpose of emulsifiers and stabilizers is mix together of different ingredients that
normally would not mix, namely fat and water. This mixture of the aqueous and lipid phases
is then maintained by stabilizers. For example, these additives are essential in the production
of mayonnaise, chocolate products and fat spreads. The manufacture of fat spreads (reduced-
fat substitutes for butter and margarine), has made a significant contribution to consumer
choice and dietary change, which would be impossible without the use of emulsifiers and
stabilizers. Other reduced and low-fat versions of a number of products are similarly
dependent on this technology.
In addition to this function, the term stabilizers is also used for substances that can
stabilise, retain or intensify an existing colour of a foodstuff and substances that increase the
binding capacity of the food to allow the binding of food pieces into reconstituted food.
Some emulsifier and a few stabilizers used in food are synthetic compounds, while many
of them are natural substances. Hygeian use of these additives need to be known and
controlled their negative effects on people health. Although almost each of them is generally
recognized as safe (GRAS), some of them still strict the level of use. Moreover, the increasing
awareness of problems with food allergy and intolerance has led to the requirement to state
the source of certain emulsifiers on food labelling. For example, lecithin derived from soya is
286 Linwei Liu and Shiyuan Dong

not suitable for an individual with an allergy to soya, so labelling of the source of the
ingredient is vital to aid in consumer choice in products safe for individuals with specific
dietary requirements.

6.2. Principle of Emulsifying and Stabilization

When two immiscible liquids, e.g., oil and water are mixed by stirrer throughly together,
there are great interface appearing between them. Because the surface tension of the interface
send up the free energy of the system greatly, the system, named as disperse system, is high
unstable and will separate water and oil quickly.
Emulsifier, an amphiphilic substance, has usually a hydrophilic head and a hydrophobic
tail. When emulsifier moleculars are adsorbed to the interface, the head and the tail will point
into the water and oil respectively, which will decrease the surface tension and increase
stability of the disperse system. If water, oil and emulsifier are mixed together, the disperse
system will formed much easier than before. That is why emulsifier has the funtion to make
and stablize emulsion and other disperse system.
Stabilisers may have a little emulsifier's function or not, but they can enhance the stability
of disperse system by other effects. It may increase the viscosity of the system, which
decrease the thermal motion speed of the droplets so as to retard their aggregation. It may be
adsorptive as the second layer on the first adsorptive layer on the surface of the droplets,
which increase the steric restriction to aggregation.
Addition from the fundamental function, emulsifier has other useful functions for food.
Formation or deformation of foaming also need emulsifier to be an accelerant for forming or
deforming the airtight film on the bubble. Another function is wetting effect, which means
moisten the solid surfaces. Solid material is mixed with an emulsifier or its surface is spread
with emulsifier, the surface then becomes hydrophilic. For example, chewing gum is apt to
stick to teeth. We can prevent adhesion by wetting the surface of chewing gum by adding
emulsifier. Emulsifier can act on starch and protein, which is important also. When emulsifier
combine with the amylose helix structure, the α-starch can not change to β-starch, which
prevent degradation of starch in bread and other millet cake, so that extending shelf life of
bread products. When making sponge dough, emulsifier can modify gluten molecules and
enhance its film-forming power, resulting in good spreadability and improvement of working
efficiency. Thus, easy-rising bread can be obtained. Emulsifiers stabilise the emulsion in low-
fat spreads providing the right stability and mouthfeel and reduce spattering in frying
margarine also.

6.3. Kinds of Emulsifiers

6.3.1. Kinds of Emulsifiers


Only food emulsifiers defined as food additives are usable to food by law. Those
emulsifiers are shown in Table 9-2. Some of the emulsifiers have been used ordinarily.
Food Additives 287

Table 9-2. Food emulsifiers

Name Common Name


Glycerin Fatty Acid Esters Monoglyceride (MG)
Acetic Acid Esters of Monoglycerides Acetylated Monoglyceride (AMG)
Lactic Acid Esters of Monoglycerides Lactylated Monoglyceride (LMG)
Citric Acid Esters of Monoglycerides CMG
Succinic Acid Esters of Monoglycerides SMG
Polyglycerol Esters of Fatty Acids PolyGlycerol Ester (PGE)
Sorbitan Esters of Fatty Acids Sorbitan Ester (SOE)
Polyoxyethylene sorbitan esters of fatty Tween
acids
Propylene Glycol Esters of Fatty Acids PG Ester (PGME)
Sucrose Esters of Fatty Acids Sugar Ester (SE)
Lecithin Lecithin (LC)

When an emulsifier will be used for a purpose, knowing its classification is important
because different classes of emulsifier have different properties, function and safety. For the
student to learn the basical knowledge, the concept of HLB (hydrophile-lipophile balance)
should be understood. The Hydrophilic-lipophilic balance of a surfactant is a measure of the
degree to decide whether it is hydrophilic or lipophilic, determining by calculating values for
the different regions of the molecule, as described by Griffin in 1949 and 1954.
Griffin's method for non-ionic surfactants as described in 1954 works as follows:
HLB = 20Mh / M
Where, Mh is the molecular mass of the hydrophilic portion of the Molecule, and M is
the molecular mass of the whole molecule, giving a result on an arbitrary scale of 0 to 20. An
HLB value of 0 corresponds to a completely hydrophobic molecule, and a value of 20 would
correspond to a molecule made up completely of hydrophilic components.
The HLB value can be used to predict the surfactant properties of a molecule:

Value from 0 to 3 indicates an anti-foaming agent.


Value from 4 to 6 indicates a W/O emulsifier.
Value from 7 to 9 indicates a wetting agent.
Value from 8 to 18 indicates an O/W emulsifier.
Value from 13 to 15 is typical of detergents.
Value of 10 to 18 indicates a solubiliser or hydrotrope.

Some of emulsifies' HLB values are listed in Table 9-3.

6.3.2. Common Used Emulsifiers


(1) Glycerine fatty acid esters.
Glycerin fatty acid esters (mono[di,tri] glycerides of fatty acids) are made from glycerin
and animal and plant oils/fats or their fatty acids. Those are generally produced by inter-
esterification method. Monoglycerides have various characteristics depending on the kind and
the content of its fatty acid. They are applied to many different fields being as an emulsifier,
foaming agent, anti-foaming agent, anti-tack agent, starch-modifying agent and anti-bacterial
agent, so you need to select the most appropriate type of monoglycerides for respective
288 Linwei Liu and Shiyuan Dong

purposes. In China, those emulsifiers are extensively used in dairy, pasta, butter, and coffee
drinks, with ML from 5 g/kg to production need.
(2) Acetic acid esters of monoglycerides.
Acetic acid esters of monoglyceride, also called as acetylated monoglyceride, are an
emulsifier in which acetic acid is bound with monoglyceride. It has little emulsifying activity
but there are many characteristics and application fields as follows: Soft acetylated
monoglyceride is able to expand by more than 8 times with tension. It is an extremely stable
oil of which peroxide value does not increase even when heated at 97.7 °C for 1000 hours.
The combination of liquid acetylated monoglyceride and hydrogenated fats can improve the
quality of fats, for example, margarine characterized with small temperature changes and
wide plasticizing range, can be produced with them. It is a liquid characterized by being less
oily even at low temperatures and available as a solvent, lubricant, etc. Although it has no
function as a good emulsifier, it is usable for foaming fats and oils by itself or in combination
with other emulsifiers because of its stable alpha-crystal structure. Practically, it is used as
powdered foaming agents, solvents, plasticizers for gums and coating agents for food.

Table 9-3. HLB values of some emulsifiers [7]

Emulsifier HLB value


Acetylated monoglycerides 1.5
Sorbitan trioleate 1.8
Glycerol dioleate 1.8
Sorbitan tristearate 2.1
Propylene glycol monostearate 3.4
Glycerol Monoleate 3.4
Glycerol monostearate 3.8
Acetylated monoglycerides (stearate) 3.8
Sorbitan monooleate 4.3
Propylene glycol monolaurate 4.5
Sorbitan monostearate 4.7
Calcium stearoxyl-2-lactylate 5.1
Glycerol monolaurate 5.2
Sorbitan monopalmitate 6.7
Soy lecithin 8.0
Diacetylated tartaric acid esters of monoglycerides 8.0
Sodium Stearoyl lactylate 8.3
Sorbitan monolaurate 8.6
Polyoxyethylene (20) sorbitan tristearate 10.5
Polyoxyethylene (20) sorbitan trioleate 11.0
Polyoxyethylene (20) sorbitan monostearate 14.9
Sucrose monolaurate 15.0
Polyoxyethylene (20) sorbitan monooleate 15.0
Polyoxyethylene (20) sorbitan monopalmitate 15.6
Food Additives 289

(3) Lactic acid esters of monoglycerides.


Lactic acid esters of monoglyceride are called lactylated monoglyceride in which lactic
acid is bound with monoglyceride (Figure 9-4). Its foaming ability is stronger than
emulsifying ability. It is used in shortening for cakes, desserts and in foaming for cream by
itself or in combination with monoglyceride. Lactic acid esters of monoglyceride are a
mixture of isomers even only one kind of fatty acid being its fatty acid constituents (see the
Figure below this paragraph). That is why these kind emulsifiers have wide HLB from 2.0 to
15. Lactic acid esters of monoglyceride are a very safe emulsifier which can be used in
appropriate level based on the need of food production according to the Chinese GB2760.
(4) Citric acid esters of monoglycerides.
Citric acid esters of monoglyceride are called citrated monoglyceride, in which citric acid
is bound with monoglyceride and the obtainable products are mixtures containing a few
monoglycerides. It is a highly hydrophilic emulsifier and a stable alpha-crystal structure used
for margarine, dairy products, such as coffee whitener and cream. It also used as an emulsion
stabilizer for mayonnaise and dressing by utilizing its strong acid-resistance.
(5) Succinic acid esters of monoglycerides.
Succinic acid ester of monoglyceride is called succinylated monoglyceride, in which
succinic acid is bound with monoglyceride. It is insoluble in cold water, dispersible in hot
water, and soluble in hot alcohol, fats and oils. Succinylated monoglyceride forms a complex
with starch which is able to react with protein. It is used as a dough modifying agent and an
emulsifier for shortening.
(6) Polyglycerol esters of fatty acids.
Polyglycerol esters of fatty acids are called polyglyceryl ester, an emulsifier in which
fatty acid is bound by esterification with polyglycerine, and generally it is dispersible in water
and soluble in oil. Its hydrophilicity and lipophilicity greatly change with the degree of its
polymerization and the type of fatty acid. Its HLB ranges from 3 to 13. It has a variety of
functions depending on these conditions, and is usable for various purposes. It is used in
many types of food as an O/W and W/O emulsifier for milk products containing acid and salt
and a modifier to control the crystallization of fats. In China, tripolyglyceryl monostearate is
used in ice-cream, bread and pastry with ML being 3.0, 0.1 and 0.1 g/kg.
(7) Sorbitan esters of fatty acids.
Sorbitan esters of fatty acid are called sorbitan ester, which is produced by esterification
of sorbitol and fatty acid. It is a mixture of sorbitol ester and sorbide ester, which are
simultaneously produced as well as sorbitan ester. There are many types of sorbitan esters
with the kinds of fatty acids and various degrees of esterification. Those are generally used as
emulsifier for cream, W/O emulsion, etc. It is applied in limited fields by itself because its
special characteristics other than emulsifying capability are few; however, it is widely used as
a major emulsifier in combination with other emulsifiers with different functions.

Figure 9-4. Sketch of two isomers of lactic acid esters of monoglyceride.


290 Linwei Liu and Shiyuan Dong

In China, sorbitan monolaurate, monopalmitate, monostearate, tristearate, and


monooleate (brand name are Sorbitan 20,40,60,65 and 80) are widely used in many foods
such as hydrogenated vegetable oil, dairy, bean products, wheaten food, and drinks with ML
from 0.5-10.0 g/kg according to the Chinese GB 2760.
(8) Polyoxyethylene sorbitan esters of fatty acids.
Polyoxyethylene sorbitan esters of fatty acids are called polyoxyethylene sorbitan ester,
which is produced by the combination of polyoxyethylene and sorbitan ester.
Polyoxyethylene sorbitan monolaurate, monopalmitate, monostearate, monooleate are called
Tween 20, 40, 60, and 80 (brand name) respectively. They are generally used as emulsifier
and stabilizer for O/W emulsion system, such as flavored milk and other liquid milks that use
non-diary ingredients. According to the Chinese FDs standard, they can be used in dairy,
drinks, bean products and bread and its ML is about 0.5-2.5 g/kg.
(9) Propylene Glycol Esters of Fatty Acids (PG ester).
Propylene glycol esters of fatty acids is called PG ester, in which propylene glycol and
fatty acid are linked by ester bonding and the product obtained from inter-esterification, is a
mixture of monoester and diester.
To isolate the monoester with surface effects, high purity product of monoester is
produced by molecular distillation and distilled monoglyceride.
PG ester has little emulsifying action, but has a tendency to keep its alpha-crystal
structure. Since it is usable as a foaming agent when combined with monoglyceride, it is used
as a powder-foaming agent for cakes and desserts, liquid shortening, etc, with the exception
of whole or partially skimmed milk. The ML is 5.0 g/kg according to the Chinese GB2760.
(10) Sucrose Esters of Fatty Acids.
Sucrose esters of fatty acids are called sucrose ester, prepared from sucrose and methyl
and ethyl esters of food fatty acids or extracted from sucroglycerides. They occur as white to
yellow-brown powdery or massive substances, or as colorless to red-brown, viscous resinous
or liquid substances. They are odorless or have a slight, characteristic odor, sparingly soluble
in water, soluble in ethanol. The solubility in oil increases with the increase of the degree of
esterification.
Because sucrose esters have many sub-categories and most of them are complex sucrose
and different fatty acids, the products of sucrose esters, generally being mixtures of different
components, have HLB ranging from 1 to 16. Owing to the wide ranging HLB, sucrose esters
have many different functions. It is used as an emulsifying and dispersing agent for foods,
such as cream, and bactericidal agents for canned coffee. The ML is from 1.5 g/kg for fats
and oils essentially free from water to 10.0 g/kg for vegetable oils and fats according to the
Chinese GB2760.
(11) Calcium Stearoyl-Lactate and sodium stearoyl lactylate.
Calcium (or sodium) stearoyl-2 lactate is called CSL. It is a product obtained by bonding
two lactic acids and stearic acid and partially neutralized with calcium (or sodium). It is a
mixture including the original stearic acid and salt. It is an anionic emulsifier having a strong
ability to bind protein and is used as a dough modifier for flour foods like bread. In China, it
also used in wheaten foods, meat enema and dairy drinks with ML being 2.0 g/kg.
Sodium stearoyl lactylate can be used as emulsifier or stabilizer for setting up water-in-
oil food emulsions containing at least 80% fat. The maximum level of use is 1.5 g/kg
according to the Chinese GB2760. Sodium stearoyl lactylate can be used as emulsifier or
stabilizer for making breads, rolls and pastries. The ML is 2.0 g/kg according to the Chinese
GB2760.
Food Additives 291

(12) Lecithin (LC).


Lecithin is a mixture containing phospholipids as the major component and widely found
in animals and plants such as soybeans, corn, rapeseed and egg yolk. It has long been used as
a natural emulsifier. The products in the market are paste lecithin or powdered lecithin of high
purity.
In the food industry, lecithin has multiple uses: in confectionery it reduces viscosity, to
replace more expensive ingredients, control sugar crystallization and the flow properties of
chocolate, improve the homogeneous mixing of ingredients and shelf life for some products,
and be used as a coating. In emulsions and fat spreads, it stabilizes emulsions, reduces
spattering during frying, and improves texture of spreads and flavor release. In doughs and
bakery, it reduces fat and egg requirements, helps even distribution of ingredients in dough,
stabilizes fermentation, increases volume, protects yeast cells in dough when frozen, and acts
as a releasing agent to prevent sticking and simplify cleaning. It improves wetting properties
of hydrophilic powders (e.g. low-fat proteins) and lipophilic powders (e.g. cocoa powder),
controls dust, and helps complete dispersion in water. It can be used as a component of
cooking sprays to prevent sticking and as a releasing agent.
For example, lecithin is the emulsifier that keeps cocoa and cocoa butter in a candy bar
from separating. In margarines, especially those containing high levels of fat (>75%), lecithin
is added as an 'anti-spattering' agent for shallow frying.
Most major religions have no dietary restrictions on the use of lecithin. In China, this
emulsifier can be used in appropriate level based on the need of food production according to
the Chinese GB2760.

7. STABILIZERS, THICKENERS AND GELLING AGENTS


7.1. Definition

All the stabilizers, thickeners and gelling agents are known and described as food
hydrocolloids implying that functional properties are obtained by mixing them with water.
They are macromolecules, such as carbohydrate polymers, proteins and their modified forms,
which are water soluble. These additives include traditional materials such as starch,
cellulose, gums, obtained from many land plants, and gelatin, an animal by-product, and
microbial polysaccharides of xanthan, gellan and pullulan, and their modified forms. Even at
low concentrations, they are able to increase a system's viscosity, to form gels and to stabilize
emulsions, suspensions or foams. These compounds are also active as crystallization
inhibitors and are suitable for aroma encapsulation, which is often needed for dehydrated
food.

7.2. Classification

Raw material origin has been used to classify hydrocolloids, for example as seaweed
extracts, seed gums, fermentation products, plant exudates, animal gelatin or microorganic
gums. Their general functional properties may also be used to classify them as thickeners,
stabilizers or gelling agents. The resourceful plant polysaccharoses, like starch and cellulose,
are derived by some chemical methods are classified modify polysaccharoses. Natural
292 Linwei Liu and Shiyuan Dong

stabilizers, thickeners and gelling agents coming from plants can be classified based on their
sources shown in Table 9-4.

7.3. Functional Properties

The following is a brief overview of the key functional properties for gelling agents,
stabilizers or thickeners used. These ingredients need to be added to commercially-produced
food to maintain their structure and physical properties during processing. For example, fruits
such as strawberries, raspberries and cherries need the addition of a small amount of pectin to
form a jam. Nutritional properties are relatively new and nutraceutical or health-enhancing
properties are even more recent. Further work is sure to advance the use of hydrocolloids
beyond modification of the rheology of foods.

7.3.1. Viscosity Adjustment


Viscosity is probably one of the most widely used properties. In this respect,
hydrocolloids are widely used to thicken sauces, soups, stews and other dishes with a high
liquid content, and the thickened water simply adds body, texture and mouth feel to a food
such as table syrups, particularly low-calorie syrups. In other cases, hydrocolloids are often
used also in systems where the oil or fat content has been reduced or eliminated through
substitution with water. The hydrocolloid thickens water, which, in turn, replaces the fat or oil
to give a product with similar properties to the full-fat food. A typical application for this
function is reduced-fat salad dressings.

7.3.2. Emulsion Stabilization


If oil or fat is partially removed from a formulation or replaced with thickened water, an
emulsion is usually formed. Often the function of the hydrocolloid is to stabilize the
emulsion, to prevent separation and, in the case of frozen foods, to control ice crystal
formation. New technology and new ingredients have been developed specifically to address
the problem of ice crystals in frozen foods, but hydrocolloids will continue to play a role.
Virtually every ice cream product sold in retail outlets is stabilized with carrageenan, locust
bean gum and/or guar gum. Low-fat salad dressings also benefit from emulsion-stabilizing
properties.

Table 9-4. Groups of plant-derived hydrocolloids [8]

Group Compounds examples


Seeds gum Guar gum, Locust bean gum, Tara gum
Plant exudates Acacia gum, Tragacanth gum
Fruits extract Pectin
Plant materials Cellulose, Starches
Seaweed extract Agar, Alginates, Carrageenan

7.3.3. Suspension Stabilization


If insoluble particles are included in the thickened product, then separation and settling
should be eliminated or at least minimized. Some hydrocolloids create solutions with a ''yield
point'' that will keep particles immobilized in suspension. Salad dressing is a good example of
Food Additives 293

this and xanthan gum is the typical hydrocolloid to supply this functionality. For beverage
with pulp suspended, like orange juice with pulp particle, precipitation can be prevented by
adding suitable amount agar to crate a small yield point which is big enough to stop the pulp
settling during the shelf-life and small enough to keep the beverage's flow property like
normal drink.

7.3.4. Gelation
One of the key texturising aspects of hydrocolloids is the ability to gel and solidify fluid
products. For example, in gelled milk desserts, even low levels of carrageenan will form a
solid milk gel. Other classic gelling agents are pectin, gelatin and agar. However, others will
form a gel under specific conditions. Certain grades of alginates form gels with calcium ions.
Xanthan and locust bean gum alone do not gel, but their combination display synergy and
form a strong cohesive gel. Methyl cellulose and hydroxypropylmethyl cellulose are unusual
in forming solutions that reversibly thicken or gel when heated. The food industry has a
myriad of gelling applications ranging from soft, elastic gels to hard and brittle gels.

7.3.5. Nutritional and Nutraceutical Function


There is already a wide use of some hydrocolloids. For example, Arabic and guar gum
are sources of soluble dietary fiber. Much research has been conducted in the nutraceutical
benefits of hydrocolloids. Potential benefits range from cholesterol reduction to cancer risk
prevention.

7.4. Common Used Stabilisers, Thickeners and Gelling Agents

The following agents are widely used in food industry. Almost every one is permitted to
be used in any food categories, except those special mentioned in appropriate level, based on
the need of production according to the Chinese GB 2760.

7.4.1. Acacia Gum


Acacia gum, also known as gum arabic, is a natural gum exudate obtained from acacia
trees in the 'African sub-Sahelian zone'. The gum is a complex mixture of polysaccharides
and glycoproteins that have a highly branched compact arabinogalactan structure which gives
a low-viscosity solution together with a central protein fraction that provides good
emulsification properties. The powder hydrates readily in water and concentrations up to 40-
50% can be handled easily. Its applications include a range of confectionery products,
flavored oil emulsions and capsules and health foods as a stabilizer and a source of soluble
fiber with prebiotic properties.

7.4.2. Agar
Agar is extracted from red seaweed (Rhodophyceae) and has been used in foods for more
than 350 years. Agar consists of a mixture of agarose and agaropectin. Agarose is a linear
polymer, made up of the repeating monomeric unit of agarobiose. Agarobiose is a
disaccharide including D-galactose and 3, 6-anhydro-L-galactopyranose. Agaropectin is a
heterogeneous mixture of smaller molecules that occur in lesser amounts. Agar is insoluble in
cold water and hydrates when boiled. Cooling solutions below about 40 °C produce very firm
brittle gels which can be melted by heating above 85 °C. Food applications include water
294 Linwei Liu and Shiyuan Dong

dessert gels, aspics, confectionery jellies, canned meats gels, a vegetarian gelatin substitute, a
thickener for soups, and as a clarifying agent in brewing.

7.4.3. Alginates
Alginates are derived from various species of brown seaweed found off the coasts of the
North Atlantic, South America and Asia. They are produced as a range of salts, but sodium
and potassium alginate are predominantly used in foods. It is a polysaccharide, consisting
primarily of D- and L-galactose units. About every tenth D-galactopyranose unit contains a
sulphate ester group. Calcium, magnesium, potassium or sodium cations are also associated
with the polysaccharide. It is insoluble in cold water and soluble in boiling water. Sodium or
potassium alginate hydrates in cold or hot water to give viscous solutions. The controlled
interaction between sodium or potassium alginate and calcium salts gives cold-setting gels
that are shear irreversible and heat stable. Control is affected using citrate or phosphate
sequestrants, or by processing at temperatures above about 70 °C and cooling. Typical food
applications include reformed foods such as onion rings and olive fillings, cold-setting bakery
cream fillings and heat-stable bakery and fruit fillings.

7.4.4. Carrageenan
Carrageenan is obtained from red seaweeds. Different species provide a number of
carrageenan extracts. Carrageenan is a hydrocolloid consisting mainly of the ammonium,
calcium, magnesium, potassium and sodium sulphate esters of galactose and 3, 6-
anhydrogalactose polysaccharides. These hexoses are alternatively linked -1, 3 and -1, 4 in
the copolymer. The relative proportions of cations existing in carrageenan may be changed
during processing to the extent that may become predominant. Depending on the source of
seaweed, the functional components are kappa-carrageenan, iota-carrageenan or lambda-
carrageenan, each of which is characterized by the content of sulphate groups and gelling
properties. Carrageenan is soluble in water of about 80 °C, forming a viscous, clear or slightly
opalescent solution that flows readily. It is insoluble in ethanol.
Kappa carrageenan and furcellaran form thermally reversible firm, brittle gels. Iota
carrageenan gives soft, elastic gels. Kappa carrageenan interacts synergistically with
polymannans, such as locust bean gum and konjac, to give strong cohesive gels. Blends of
kappa with iota carrageenan or polymannans are used to give a range of gel textures used in
injected meats, canned meats and water dessert gels..
Hybrid (kappa/iota) carrageenan is particularly suitable for firm, creamy textures in dairy
desserts. Lambda carrageenan is non-gelling but thickens in drinks and dairy desserts. A
specific interaction between kappa carrageenan and kappa casein is widely used to stabilise
dairy products including milk beverages, ice cream mixes and processed cheese products.

7.4.5. Microcrystalline Cellulose


Microcrystalline cellulose is produced by converting fibrous cellulose to a fine-particle
form crystalline cellulose using acid hydrolysis. This material can be readily dispersed in
water using high shear. This dispersion will reconstitute to deliver a colloidal form which is
unique when compared to other soluble food hydrocolloids. It exhibits a variety of desirable
characteristics including suspension of solids, heat stability, ice crystal control, emulsion
stabilization, foam stability, texture modification and fat replacement. Food applications
include frozen desserts and ice cream, whipped toppings, low-fat mayonnaise and salad
Food Additives 295

dressings, ambient-stable dairy and non-dairy beverages, nutritional drinks, ready-to-use


bake-stable bakery fillings and fruit fillings and dairy and non-dairy creams.

7.4.6. Cellulose Derivatives


Cellulose derivatives are substances commonly made from cellulose by chemical
modification. The common used cellulose gum in food is carboxymethyl cellulose and
hydroxypropylmethyl cellulose. In food applications, cellulose gum is an effective thickener
and moisture binder used for clear beverages, dairy products, such as ice cream, bakery and
other prepared foods. Hydroxypropylmethyl cellulose exhibits thermogelation, utilized for
bake-stable sauces, fillings and formed foods. Hydroxypropylmethyl cellulose is also a
surface-active agent to be used to stabilize foams and emulsions.

7.4.7. Starch Derivatives


Starch derivatives are wide substances commonly made from starch by chemical or
phsical modification. The common used starch derivatives in food are starch acetate, distarch
phosphate, hydroxypropyl distarch phosphate, acid treated starch, oxidized starch, oxidized
hydroxypropyl starch, acetylated distarch phosphate and acetylated distarch adipate. Starch
derivatives are easier gelatination and more anti-aging than common strarch. The viscosity of
their solution is also more stable than the common starch. Starch derivatives can be used in
any food to substitute starch to make the food more stable.

7.4.8. Gelatine
Gelatine is a protein material obtained from animal connective tissue using hydrolysis in
acidic (type A) or basic (type B) solution followed by hot water extraction. Gelatine hydrates
readily in warm or hot water to give low-viscosity solutions that have good whipping and
foaming properties. Concentrated solutions containing up to 40 % gelation are made for use
in confectionery. After cooling, the network of polypeptide chains associates slowly to form
clear, elastic gels that are syneresis free. The thermoreversibility of gelatine gels gives them
unique properties: the melting point is below 37 °C so they melt in the mouth to give smooth
textures with excellent flavour releasing. Gelatine is used in a wide range of food,
pharmaceutical and photographic applications. Food products include jelly candies and
aerated confectionery, yogurt and other cultured desserts, dairy desserts and creams, low-fat
spreads, canned meat products, water dessert gels and decorative aspics.

7.4.9. Seed Gums


The origin of seed gums, such as guar gum and carob or locust bean gum, are produced
by removing the outer coating of the seed and grinding the endosperm. The composition and
structure of the galactomannans are described and linked to their functional properties. Seed
gums are very effective thickening agents in water. In addition, guar gum interacts
synergistically with xanthan to increase viscosity. Locust bean gum consists mainly of high
molecular weight (approximately 50,000-3,000,000) polysaccharide composed of
galactomannans. It is soluble in hot water and insoluble in ethanol. Carob gum forms
cohesive, elastic gels with xanthan gum and it increases the strength and elasticity of kappa
carrageenan and agar gels. The main applications are for thickening convenience foods, dairy
products, including frozen products such as ice cream, soft drinks and fruit juices, bread and
pastry, fruit preserves, baby food, instant products including puddings, flans and pudding
powder, and for dietary fibre in baked goods and pet foods.
296 Linwei Liu and Shiyuan Dong

7.4.10. Pectin
Pectin is a polysaccharide that is naturally present in most land plants, although
commercial pectin is primarily extracted from citrus peel and apple pomace. Two forms of
commercial pectin are available: high methyl- and low methyl-esterified pectin; and two
versions of the latter exist: a conventional and an amidated form. High methyl-esterified
pectin forms gels in high soluble solids and acidic systems, whereas low methyl-esterified
pectin forms gels in a much broader pH and soluble-solids range, but requires the presence of
divalent cations for gelling. As a consequence, each type has its own particular function.
Nevertheless, general attractive features include excellent flavor release, good processing
characteristics and stability at low pH. Its traditional and major function is to act as a gelling
agent in foods, but, nowadays, it also serves as a thickening and stabilizing agent. The
application of pectin is diverse and covers fruit-based products, dairy products, acidified milk
drinks and other beverages, confectionery, bakery products, various fine foods and spreads.
Additionally, pectin is used in the pharmaceutical industry. Finally, increasing consumer
awareness of healthy life-style habits and the emerging trend to produce functional foods
increases the significance of the status of pectin as a water-soluble dietary fiber.

7.4.11. Xanthan Gum


Xanthan gum is a high-molecular-weight polysaccharide of which D-glucose and D-
mannose are the dominant hexose units, along with D-glocuronic acid and pyruvic acid,
secreted by the microorganism Xanthomonas campestris and produced commercially in a
batch fermentation process. It exists as the sodium, potassium or calcium salts. Its solutions
are neutral. It is soluble in water, and insoluble in cold water. The solution is a neutral and
viscous one with pseudoplastic flow behaviour. This gives excellent suspension and cling at
low shear and excellent mouthfeel and pouring properties at high shear. The xanthan gum
molecule has a cellulosic backbone with side chains that wrap around the backbone protecting
it and conferring excellent stability across a wide pH range and tolerance of high salt
concentrations and ingredients, such as glycerol and alcohol. The rigid backbone helps to
maintain viscosity during heating. Xanthan gum shows synergistic thickening with guar gum
and forms very elastic cohesive gels with locust bean gum and konjac mannan. Typical
applications include sauces and dressings, baked goods, beverages, desserts and ice creams.

7.4.12. Gellan Gun


Gellan gum is a fermentation polysaccharide produced by the microorganism
Sphingomonas elodea. It has a straight chain structure based on repeating glucose, rhamnose
and glucuronic acid units with side groups of acryl groups. Gellan gum hydrates in hot water
and the low-acyl form also hydrates in cold water with sequestrants. On cooling, native high-
acyl gellan gum gives gels that are soft and elastic. Low-acyl gellan gum gels at very low
concentrations using both monovalent and divalent cations gives firm, brittle textures with
excellent thermal stability. Combinations of the two kinds of gellan gum can be used to
control syneresis and form a range of textures from soft and elastic to firm and brittle. A
major food application is water dessert gels, particularly for Asian desserts. Other significant
applications include confectionery, dairy desserts and bakery fillings. At levels too low to
form a demoldable gel, gellan gum can form fluid gels that can suspend particulates in sauces
and dressings and fruit pulp in beverages.
Food Additives 297

7.4.13. Konjac Glucomannan


Konjac flour yields a high molecular weight, viscous polysaccharide: konjac
glucomannan. Glucomannan is a highly soluble, neutral plant polysaccharide that has been
used as a gelling agent for 2000 years in China. It gels alone or in association with other
polysaccharides where it shows strong synergistic effects in viscosity, gel strength and
elasticity. Konjac has a greasy mouthfeel and chewy texture that resemble fat. It is resistant to
digestion and has a very low calorific value. As well as being a healthy food, it is a preferred
texturing ingredient in Asia. Glucomannan has generally recognised as safe (GRAS) status in
the USA and it is well established as an additive in the EU food industry. Cost-effective as a
thickener, the effects on post-prandial sugar and lipid levels, low digestibility and prebiotic
fermentation have placed glucomannan amongst the most exciting hydrocolloids on the
'emerging' world nutraceutical market.

7.4.14 Tragacanth Gum


Tragacanth is a dried exudation obtained from the stems and branches of Astragalus
gummifer Labilliardiere and other Asiatic species of Astragalus (family
Leguminiosae).Tragacanth consists mainly of high molecular weight polysaccharides
(approximately 800.000) i.e. galactoarabans and acidic polysaccharides, which are
hydrolyzed to galacturonic acid, galactose, arabinose, xylose, and fucose. Small amounts of
rhamnose and glucose (derived from traces of starch and/or cellulose) may be present. Gum
tragacanth is a viscous, odorless, tasteless, water-soluble mixture of polysaccharides. It is
used in pharmaceuticals and foods as an emulsifier, thickener, stabilizer, and texturant
additive.

7.4.15. Chitin and Chitosan


Chitin and chitosan have the similar chemical structure. Chitin is made up of a linear
chain of acethylglucosamine groups. Chitosan is obtained by removing enough acethyl groups
(CH3-CO-) for the molecule to be soluble in most diluted acids. This process, called
deacetylation, releases amine groups (-NH) and gives the chitosan a cationic characteristic.
This is especially interesting in an acid environment where the majority of polysaccharides
are usually neutral or negatively charged. Chitin can use in non-dairy creamer, ice creams,
jams, marmalades, peanut butter, sesame paste, mayonnaise, lactobacillus drink, and beer
with ML 0.4-5 g/kg based on GB2760. Chitosan can be an effective complement to help lose
weight during diet period or to stabilise ones weight, and used as clarifier in wine industry.

7.4.16. Others
Polydextrose, propylene glycol alginate, fenugreek gum, sesbania gum, tamarind seed
polysaccharide gum, sodium carboxymethyl starch, ablmoschus manihot gum, artemisia gum,
gleditsia sinensis lam gum, linseed gum, polyglycerol esters of fatty acid, hydroxypropyl
starch phosphated distarch phosphate, methyl cellulose and ethyl cellulose are other
thickeners, stabilisers and gelling agents used in food in China. They are used in some
appointed food categories with limited amount as ML in GB2760.
298 Linwei Liu and Shiyuan Dong

8. PHOSPHATE AND POLYPHOSPHATES


Phosphates' buffering capacity, chelating metal ions, water-holding capacity, and
interacting with long chain polyelectrolytes such as protein, make them widely used as FDs.
Phosphates are one of the most widely used functional food ingredients. Applications of
the various salt forms of phosphates, such as leavening agents in baked goods, moisture loss
inhibitors in frozen and processed meats, emulsifiers in dairy products, and buffering agents
for many different food formulations, are included. For example, monocalcium (MCP) and
dicalcium phosphate dihydrate act as leavening agents in baked goods. MCP is also useful as
a dough conditioner, and can be used to strengthen the gel formation in instant pudding
products. Tricalcium phosphate is very useful in dry powder mixtures, to prevent the
adsorption of moisture and allow the powders to flow properly. When these phosphates are
used as functional food ingredients, they also provide the fortification of calcium and
phosphorus.
Phosphates are used in pasteurized cheese products, ice cream, frozen custard, breads,
rolls, buns, flour, macaroni products, fruit jellies, preserves and jams, frozen eggs, vanilla
powder, as well as many other foods.
Polyphophates are added to flesh foods, such as meat, fish and poultty, in order to
increase the retention of water and the solubility of proteins to improve texture.
A range of phosphoric acids and their sodium potassium, and calcium salts, are used as FDs,
from the simple acid orthophosphoric acid, H3PO4, to complex polyphosphates are permitted
for use in food. Because too much of them in diet leads to loss of calcium in bones and onset
of osteoporosis, almost every kind of those FDs are used with special limited ML for different
food catogries according to the GB2760, and is banned in organic foods and drinks in some
countries.

8.1. Phosphoric Acid

Phosphoric acid (other names: orthophosphoric acid.) can only be obtained pure in the
crystalline state and slowly undergoes dehydration to diphosphoric acid. Phosphoric acid is
added to food to enhance the antioxidant effects of other compounds present, and also as an
acidity regulator. Typical products include carbonated beverages, processed meat, chocolate,
fats and oils, beer, jam and sweets. Phosphoric acid is a highly acidic ingredient in cola
drinks.

8.2. Sodium Phosphate

(1) Monosodium phosphate is a sodium salt of phosphoric acid. It is added to food to act
as an antioxidant synergist, a stabiliser and a buffer. Typical products include processed meat
products, processed cheese products. It is also used in bread, rolls, buns, artificial sweetened
fruit jelly, canned potatoes, canned sweet peppers, and canned tomatoes and as a jelling agent.
No food safety problems have been shown to occur with this chemical at the levels commonly
used in foods, but high intakes may upset the calcium/phosphorus equilibrium.
(2) Disodium phosphate is a sodium salt of orthophosphoric acid and is used as an
antioxidant synergist, stabiliser and buffering agent in food. It is also used as an emulsifier in
Food Additives 299

the manufacture of pasteurised processed cheese. Disodium phosphate is added to powdered


milk to prevent gelation. Typical products include processed meat products, processed cheese
products, powdered milk.
(3) Trisodium phosphate is the sodium salt of orthophosphoric acid and used as an
antioxidant synergist, stabiliser and buffering agent in food. Typical products include
processed meat products, processed cheese products.

8.3. Potassium Phosphates

(1) Monopotassium phosphate is a potassium salt of phosphoric acid used as an


antioxidant synergist, buffer and emulsifier in food. Typical products include sauce and
dessert mixes, jelly products.
(2) Dipotassium phosphate is a potassium salt of phosphoric acid used as an antioxidant
synergist, buffer and emulsifier in food. Typical products include cooked and other cured
meats, milk and cream powders, drinking chocolate.
(3) Tripotassium phosphate is a potassium salt of phosphoric acid used as an antioxidant
synergist, buffer and emulsifier in food. Typical products include cooked and other cured
meats, milk and cream powders, drinking chocolate.

8.4. Calcium Phosphates

(1) Monocalcium phosphate is commercially available in the anhydrous or monohydrate


form. Both are used as a leavening acid to replace cream of tartar in foods. Mono-calcium
phosphate is used extensively in the fertiliser industry. Typical products include self-raising
flour, baking powder, cake and pastry mixes, cakes and other pastry products as a baking
agent.
(2) Dicalcium phosphate - Manufactured from phophoric acid, dicalcium phosphate is
used as an antioxidant in food, as well as being a firming agent. It is available in the
anhydrous or dihydrate forms. Typical products include tinned and packaged fruit deserts,
granular food products. In the canned products it provides calcium which has been shown to
maintain the firmness of fruits and vegetables during the canning process.
(3) Tricalcium phosphate is added to table salt, sugar, baking powder and fertilisers to
give a 'free-flowing' quality. Typical products include salt, sugar and other granular foods,
packet sauce mixes, cake mixes etc.

8.5. Sodium Triphosphate

Sodium triphosphate is beside potassium triphosphate, one of the polyphosphates that are
allowed as FDs. It is used in food for soften water, as emulsifying salt for process cheese or
for the preparation of cooked sausages, surimi or fish fingers. It has antimicrobial function
also.
300 Linwei Liu and Shiyuan Dong

9. COLORANTS
Dyes and pigments used in the food are known collectively as food colours, or more
tersely, simply as colours. They are added to foods to make it more attractive to the purchaser
or consumers or to replace natural colours lost during the food processing. Canned
strawberries, for example, would be grayish-brown if colours are not added and canned peas
would be brownish green without colouring with green colorant.
In China, only 56 colorants are permitted for use as FDs. Most of them are natural, and
include saffron, cochineal, carotenes (and the closely related xanthophylls) and anthocyanins.
Other permitted colorants include synthetic or semi-synthytic coumpound, such as carotene
structure identical, sodium copper chlorophyllin and erythrosin, and inorganic substances,
such as the white pigment titanium dioxide and finely divided aluminum, silver and gold
which are used in cake decoration.
The list of permitted colours includes 9 synthesis dyes and 47 natural pigments or
extracts. Most of them are used in foods with strictly limits. For example, colouring matter
must not be added to raw meat, fish, poultry, fruit, cream, milk, honey, vegetables, wine,
coffee, tea and condensed or dried milk in most countries. Also, by agreement with the
manufacturers, colours are not used in foods made for babies and infants. The 9 synthesis
colours use in China are amaranth, ponceau 4R, erythrosin, new red, lemon yellow, indigo
blue, sunset yellow, brilliant blue and crimson. Sodium copper chlorophyllin is a semi-
synthesis colour which is also permitted in China. The 47 natural colours are β-carotene
(zymotechnics), bect-root red, turmeric yellow, curcumin, safflower yellow,
http://www.chinaadditives.com/Carthamus-Red.htm lac dye, cowberry red, papika extract
include capsorubin and capsanthin, chilli orange, caramel, carthamus yellow, gardenia
yellow, coreopsis yellow extract, wild groundnut red, broomcorn red, maize yellow include
zeaxanthin and crtotixabthin, radish red, cacao husk pigment, ang-kak, monascorubin,
vinespinach red, black currant red, neutral mustard red, gardenia blue, hippophae
rhamnoides(sea backthern) yellow, hibiscus sabaariffa(roselle) red, acorn husk brown, NP red
(NP), tanoak brown, mulberry red, natural amaranth, cherokee rose brown, wide jujube skin
extract, peanut underwear red, grape skin red, uguisukagnra color, spirulina blue, vegetable
carbon black, pale butterflybush flower yellow, gromwell red, theaflavin, tea green pigment,
citrus yellow, bixin, cochineal red and iron oxide red or black.
All of the 9 synthesis colours and most of natural colours used in China are water soluble
and purchased in a powder format that exhibits coloring solutions when they are dissolved.
Therefore, their using are very easy.
Aluminum lake pigments, made of aluminum and colorants, are water insoluble material
but oil dispersible (but generally not oil soluble) and thus can be mixed with oils and fats.
They can also be dispersed in other carriers such as propylene glycol, glycerin and sucrose
(water and sugar). Aluminum lake pigments are used in a variety of applications: (1) To color
a fat based product, such as chocolate or compound coatings. For these, we produce a
concentrated dispersion in a high quality and very stable vegetable oil. The dispersion is
added directly to the chocolate to dye it accordingly. (2) For "hard panning" (to dye the
outside of a product such as a gum ball or a pill). In this case, we produce a dispersion usually
using sucrose (sugar and water) that is applied to the candy or food as it is being tumbled and
dried. Multiple layers are applied to produce the desired shade.(3) Lakes tend to resist
bleeding. Dyes have a tendency to "bleed", or migrate from one part of the product to another.
This can be a problem in candy canes or any product where there are defined borders such as
Food Additives 301

the food with desired stripes. While Dyes are normally used in hard candy, lakes are
sometimes substituted if bleeding is a problem.

10. FOOD FLAVORANTS AND FLAVOUR MODIFIERS


10.1. Food Flavorants

Flavourings, or flavors, spices and essence, are substances or food flavor additives used
to give taste and/or smell to food. There are different types of flavourings, such as natural,
natural-identical or artificial flavouring substances, flavouring preparations of plant or animal
origin, processing flavourings which evolve flavour after heating and smoke flavourings. The
expression "natural flavouring" may only be used for flavouring substances or flavouring
preparations which are extracted from vegetable or animal materials. Generally, different
flavors are specially formulated for various food items to make them more delightful.
Flavorants contains many flavour compounds. Only the Generally Recognized As Safe
flavour compounds can be used as food flavor additives. Flavour compounds often classify
into 18 groups, shown in Table 9-5. Each of the 18 groups contains substances that have
similar chemical structure at least partly, but have not similar odors mostly.

Table 9-5. Groups of flavouring substances

Group Chemical substances


1 Isothiocyantaes (except for those generally recognized as toxic).
2 Indole and its derivatives.
3 Ethers.
4 Esters.
5 Ketones.
6 Fatty Acids.
7 Aliphatic Higher Alcohols.
8 Aliphatic Higher Aldehydes (except for those generally recognized as toxic)
9 Aliphatic Higher Hydrocarbons (except for those generally recognized as toxic)
10 Thioethers (except for those generally recognized as toxic)
11 Thiols(Thioalcohols) (except for those generally recognized as toxic)
12 Terpens
13 Phenol Ethers (except for those generally recognized as toxic)
14 Phenols (except for those generally recognized as toxic)
15 Furfural and its derivatives (except for those generally recognized as toxic)
16 Aromatic Alcohols
17 Aromatic Aldehydes (except for those generally recognized as toxic)
18 Lactones (except for those generally recognized as toxic)
302 Linwei Liu and Shiyuan Dong

10.2. Flavour Modifiers


A flavour modifier is a substance which is capable of enhancing or modifying the taste or
odour, or both, of a food. Flour modifiers or enhancers are themselves practically tasteless but
they intensify the flavour of soups, meat and other savoury foods.
The most widely used flavour enhancers are monosodium Glutamate (MSG) and sodium
5-ribonucleotide.
MSG is present in most dehydrated soups, stock bouillon cubes and other meaty prepared
foods. Some people have an alergy to MSG, which has come to be known as the "Chinese
restaurant syndrome", if they eat food containing it. This manifests itself in a variety of ways
including palpitations, chest or neck pain and dizziness. The cause is not known and the ill
effects soon disappear. It has also been reported in potter, N.N. (1986) of brain damage
resulting when MSG was injected under the skin of young mice. Though MSG is not thought
to be harmful according to Fox B.A. and Cameron, A.G (1989), precautionary measures are
still been taken for not adding it to babies and infants food.
Sodium 5'-ribonucleotide is another powerful permitted flavour enhancer.
Ribonucleotides are compounds formed from the sugar ribose, phosphoric acid and an
organic base such as guanine. They occur in all animal tissues and are present in yeast
extracts and contribute substantially to their characteristic meaty taste.
Ribonucleotide flour enhancers are used in soups, meat and fish pastes, all types of
canned meat products, sausages, meat pies and other processed food products of which the
main ingredients is meat or fish. Their flavour enhancing power is 10 times greater than that
of MSG.
Hydrolyzed proteins, broken apart into amino acids, are used by the food industry to
enhance flavor. The chemical breakdown of proteins may result in the formation of free
glutamate that joins with free sodium to form MSG. In this case, the presence of MSG does
not need to be disclosed on labeling. However, labeling is required when MSG is added as a
direct ingredient.

REFERENCES
[1] Salant, A. Nonnutritive sweeteners. In: Furia, TE. Handbook of Food Additives.
Cleveland: CRC Press, 1972.
[2] Luo, AS; Chun, Z; Luo, AX; Fan, YJ; Ge, SR. The survey and progress of the food
preservative. China Food Additives, 2005, 55-57.
[3] Wedzicha, RL. Chemistry of Sulphur Dioxide in Foods. 1st edition. London: Elsevier,
1984.
[4] Chichester, DF, et al. Antimicrobial food additives. In: Furia, TE. Handbook of Food
Additives. Cleveland: CRC Press, 1972.
[5] Zsolt, F, et al. A synthetic and in silico study on the highly regioselective Diels-Alder
reaction of the polyenic antifungal antibiotics natamycin and flavofungin. Tetrahedron
Letters, 2010, 51, 4968-4971.
[6] Ma, LR; Zhang, ZL; Li, H. New type food preservative-NISIN. Journal of Hebei
Academy of Sciences, 2001, 18, 180-182.
[7] Griffin, WC. Classification of Surface-Active Agents by HLB. Journal of the American
Oil Chemists' Society, 1949, 311.
Food Additives 303

[8] Ren, P; R, YW; Wang, LD. Properties of Plant Gum and Its Application in Food
Industry. Food Research and Development, 2004, 39-44.
In: Food Chemistry ISBN: 978-1-61942-125-7
Editors: D.Wang, H. Lin, J. Kan et al. © 2012 Nova Science Publishers, Inc.

Chapter 10

TOXICANTS IN FOODS

Wang Dongfeng1, Guoqing Huang2 and Shuhui Wang3


1
College of Food Science and Engineering, Ocean University
of China, Qingdao, China
2
College of Food Science and Engineering, Qingdao
Agricultural University, Qingdao, China
3
College of Agriculture - Ginn College of Engineering,
Aubum University, Auburn, AL, US

ABSTRACT
Microorganisms, plants and animals have evolved multiple defense systems against
predators. Among the systems, the chemical defense system is of special interest for the
food industry. Some of these compounds are toxic to specific microorganisms, insects,
birds, and mammals, but most of those are toxic to humans and can give rise to chronic or
acute symptoms of poisoning. Though some of the compounds occur in very low
concentrations, the compounds have a cumulative harmful effect if they constitute a part
of the normal diet over a prolonged period. Besides, food materials might be
contaminated by exogenous toxins. Intrinsic toxins and contaminants have been
recognized as the major components that affect food safety. This chapter introduces the
occurrence and properties of the most important toxicants in foods, including allergens,
toxic glycosides, toxic amino acids, lectins, saponins, toxins in aquatic organisms,
various antinutrients, heavy metals, pesticide residues, dioxins, veterinary and fish drug
residues, benzopyrenes as well as heterocyclic amines. Besides, measures for preventing
the generation of some toxicants are proposed.

1. INTRODUCTION
According to structure and biological effects on human body, toxicants in foods are
divided into toxic substances, harmful substances, and antinutrients. A toxic compound shows
its toxicity even in a very low dose, and a harmful substance is toxic only when its content
exceeds a certain threshold. An antinutrient may interfere in or inhibit the absorption of other
nutrients in foods. The definitions of these terms, however, are subject to changes due to the
306 Wang Dongfeng, Guoqing Huang and Shuhui Wang

improvement of analysis techniques and the availability of more data on their biological
effects. It has been found that some toxicants are beneficial in certain contents and some
substances are toxic or hazardous only in certain cases. For example, phenols in foods are
often recognized as antinutrients since they inhibit the adsorption of proteins. However,
phenols have the anti-oxidation and free radical scavenging capabilities and can be used as
natural antioxidants in foods. In addition, some well-known nutrients may be risky to certain
individuals, such as lactose, which may be deleterious to lactose-intolerant individuals.
According to origin, hazards in foods can be divided into inherent toxicants (or
endogenous toxicants) and contaminants (or exogenous toxicants). Endogenous toxicants are
metabolites produced via biosynthesis by food organisms under normal growth or metabolites
produced via biosynthesis by food organisms that are stressed. Contaminants refer to
toxicants that directly contaminate foods, toxicants that are absorbed from the environment by
food-producing organisms, toxic metabolites produced by food organisms from substances
that are absorbed from the environment, and toxicants that are formed during food preparation
[1].

Table 10-1. Endogenous toxicants in foods

Toxicants Source Effects on human upon ingestion


Glucosinolates Cruciferae seeds, oilseeds, Thyroid enlargement, thyroxine
(goitrogens) mustard seed, kale, radish, synthesis decrease, metabolism
cabbage, peanut, soybean, impairment, reduced iodine adsorption
cassava, onion, et al. and protein digestion
Cyanogenic Cassava, sweet potato, nuts, Cellular inspiration block, stomach
glycosides kidney bean, limabean, millet, upset, sugar and calcium transfer block,
broomcorn millet, et al and iodine uptake interference
Lectins Papilionaceae, cereals, soybean, Intestinal epithelial cells damage,
and other beans nutrient adsorption retardation, enzyme
inhibition, B12 and lipid adsorption
impairment, et al.
Glycoalkaloids Potato, tomato, and other Cholinesterase inhibition, stomach
immature fruits upset, hematolysis, kidney function
impairment
Gossypol Cottonseed Metal binding, iron adsorption
decrease, enzyme inhibition
Tetrodotoxin Globefish Neuron paralysis, dyspnoea and even
death
Shellfish Mussels, Mytilus edulis, scallops, Nerve paralysis and liver poisoning
toxins Meretrix meretrix, et al.
Histamine Scomber japonicus, tunny, Allergy, including blush, dizziness, and
sardine, et al. tachypnea
Mycotoxins Toxic mushrooms Gastroenteritis, nerve system injury,
hematolysis, visceral parenchymal
injury
Toxicants in Foods 307

Table 10-2. Endogenous antinutrients in foods

Antinutrient Source Effect on human upon ingestion


Interference in the adsorption of Ca,
Beet root, spinach, celeriac, Rhubarb,
Oxalic acid Fe, Zn and other metals and Ca
amaranth, tomato
metabolism

Vegetables, fruits, grape wines, cereals, Block of thiamine adsorption and


Phenols
soybeans, potato, tea, et al. decrease of metal bioavailability

Decrease of the bioavailability of Ca,


Papilionaceous plants, cereals, and the
Phytates Mg, Fe, Zn, Cu, and other metals and
seeds of all plants
that of proteins and starch
Papilionaceous seeds, peanut, cereals,
Protease Inhibition of trypsinase, chymotrypsin,
rice, maize, potato, apple, sweet potato, et
inhibitors glycopeptidase, and amylase
al.
Complexation with proteins and
Papilionaceous plants, spinach, lettuce, lipoids, hematolysis, and
Saponins
sugarbeet, soybean, tea, peanut gastroenteritis
Note that most saponins are nontoxic.
Inhibition of pancreatic enzymes,
Plant-derived foods, such as most fruits,
Tannins bioavailability decrease of thiamine,
tea, and coffee
proteins, cobalamin, and Fe
Non-protein Interference with protein metabolism,
Legume, seaweed
amino acids bone and nerve poisoning

Table 10-3. Exogenous toxicants in foods

Hazards Source Effect on human upon ingestion


Heavy metals Plants and animals Inappetency, gastroenteritis, insomnia, dizziness,
cultivated in polluted muscle soreness, anemia, and many other symptoms
rivers, estuaries, and
soils
Organophosphoru Foods polluted by Enzyme inhibition, muscle vibration, spasm, myosis,
s pesticides organophosphorus blood pressure elevation, heartbeat acceleration,
pesticides dyspnea, pneumonedema, and coma

Veterinary drugs Diary and meat products Allergy, drug-resistance development,


microecological environment change, early maturity,
physiological disorder, chronic disease symptom
exacerbation
Microbial toxins Microorganism-polluted Multiple adverse effects
foods and materials

It should be noted that all chemicals are potentially toxic, and that the dose alone
determines whether or not a toxic effect occurs. Indeed, toxicant (or toxin) is recently defined
as a ―substance that has been shown to present some significant degree of possible risk when
consumed in sufficient quantity by humans or animals‖ [2]. For example, pure water drunk to
excessive can induce an electrolyte imbalance and lead to death, which is called the ―water
toxicity‖ that is due in part to decreased renal capacity to excrete water [3]. The innumerable
308 Wang Dongfeng, Guoqing Huang and Shuhui Wang

naturally occurring chemicals in food are potentially capable of inducing toxic effects, but
few are ordinarily present in sufficient concentrations to actually show toxic effect [2, 4].
Hence, we should distinguish between ―toxic substance‖ and ―toxic effect‖. The most
frequently occurring toxicants in foods are listed in Table 10-1~10-3.
A substantial number of toxic substances have been isolated and identified This chapter
concerns only the most important toxicants that have been identified in foods.

2. ENDOGENEOUS TOXICANTS
Endogeneous toxicants include any substances produced by food materials (either plants
or animals) that are harmful to human beings. Lectins, saponins, and toxic peptides mentioned
above are endogenous toxicants.

2.1. Allergen

An allergen refers to any substance that can cause an allergy. Most immune responses of
caused by food components are IgE-mediated immediate allergic reactions. Upon exposure to
allergens, the lymphatic system of human body acts to avoid or alleviate the adverse effects.
Most antigens can be metabolized to monosaccharides, amino acids and lower fatty acids
after antigen-antibody reactions. However, complete-antigens, which are characteristic of
both immunogenicity and immunoreactivity, can cause immune reactions.
The intake of allergen-containing foods can cause allergic symptoms, such as pruritus,
functional gastrointestinal disorder and fever. Many foods have been found to contain
allergens, but the sensitivity to these foods varies with local diet, environmental factors and
possibly genetics. Epidemiological investigations reveal that the incidence of food-induced
allergy in infants and children is higher than that in adults and the incidence decreases as age
increases.
Food allergens share the following characteristics:

1. Allergens are present in most foods, such as milk, eggs, and soybean, of which, milk
and egg are the most important source of allergens. Though allergens occur widely in foods,
90% of food allergies are induced by only a small number of foods.
2. Only few food components are allergenic. For example, up to 23 types of
glycoproteins have been identified in white egg, but only ovalbumin, conalbumin, and
ovomucin are allergic.
3. The allergenicity of foods is subject to change after processing. For example, heat
treatment inactivates most allergens and the increase of acidity or the presence of digestive
enzymes reduces the allergenicity of foods.
4. Sensitive consumers could suffer allergic cross reactions. Some proteins contain the
same antigenic determinant and these proteins can cause allergic cross reactions. For instance,
at least 50% of patients allergic to cow‘s milk also react to goat‘s milk and patients allergic to
chicken eggs might be allergic to the eggs of other birds.
Toxicants in Foods 309

2.2. Toxic Glycosides

2.2.1. Properties of Toxic Glycosides


Toxic glycosides are also known as cyanogenic glycosides. These compounds often
contain glucose or rhamnose as their sugar components and are widely present in cassava,
sweet potato, nuts, kidney bean, limabean, millet, and many other cereals. Excessive intake of
toxic glycosides may lead to gastrointestinal malaise and affect the transfer of sugars and
calcium. The contents of toxic glycosides in plants vary with species and cultivation
technologies. Tables 10-4 and 10-5 list the distribution of toxic glycosides and their contents
in various cereals.
Glycosides can be hydrolyzed by enzymes to produce thiocyanates, isothiocyanates, and
perthiocyanates. All these products are toxic and can cause goiter.

Table 10-4. Distribution and hydrolytic products of some cyanogenic glycosides


in food plants [5]

Glycoside Source Hydrolytic products


Almond, cherry, peach, plums,
Amygdalin HCN, gentobiose, and benzaldehyde
apples
Dhurrin Sorghums HCN, glucose, and hydroxybenzaldehyde
Linamarin Linseed, clovers, cassava, lima beans HCN, glucose, and acetone
Lotaustralin Linseed, cloves, cassava, lima beans HCN, glucose, and 2-butanone
Prunasin Cherry, almond HCN, glucose, and benzaldehyde
Cicianin Vetches HCN, vicianose, and benzaldehyde

Table 10-5. Thiocyanate contents in typical vegetables


(mg/100g edible part of fresh leaf)

Plant Content
Cabbage 3~6
Savoy 18~31
Brussels sprout 10
Cauliflower 4~10
Kidney bean, turnip <1
Kohlrabi 2~3
Colza 2.5
Swedish turnip 9
Lettuce, Spanish, onion <1

Figure 10-1. Structure of glucosinolates; side group R varies.


310 Wang Dongfeng, Guoqing Huang and Shuhui Wang

Figure 10-2. Hydrolysis of glucosinolates by myrosinase.

2.2.2. Glucosinolates

Structure and main species

Glcosinolates, also known as β-thioglucoside N-hydroxysulfates, contain one branch and


one glucopyranose residue bound via a sulfur atom, as shown in Figure 10-1.
The side group R may be sulfur-containing groups, straight-chain alkanes, side-chain
alkanes, olefins, saturated alcohols, ketones, aromatic groups, benzoates, indole, polyglucose
groups, or others. Among the glucosinolates that have been identified, one third of the
compounds possess sulfur-containing R groups and the sulfur atoms are present in multiple
forms, such as methyl sulfanes, methyl thionyl alkanes, and methyl sulfuryl alkanes. Plants of
the Cruciferae family have high levels of glucosinolates and the side chain R may be alkanes,
ω-methyl sulfanes, aromatic groups, or heterocyclic groups.
Glucosinolates that act as progoitrins are the source of organic nitriles, isothiocyanates,
and SCN ions. Some of these compounds have been shown to be quite harmful if consumed
in sufficient amounts by humans and animals.

Enzymatic hydrolysis and changes in processing

Glucosinolates are quite stable. They are the precursors of isothiocyanates and are present
in much higher levels than their hydrolyzates in fresh plants. Glucosinolates can produce a
large variety of products when exposed to myrosinase as shown in Figure 10-2. In the case of
fresh plants chewing or tissue injury during cultivation, harvest, transport, and processing due
to scratching or freezing-thawing, glucosinolates get contact with myrosinase (EC 3.2.3.1)
and are hydrolyzed to toxic isothiocyanates.
Glucosinolates and their enzymatic hydrolyzates are water soluble. During cooking, more
than a half of the compounds are released to water, but the specific loss caused by processing
varies with vegetable species.
Myrosinase can be activated by Vc. Many facts have proved that myrosinase is inactive
in the absence of Vc. The activation is not due to the reducing property of Vc, instead due to a
nucleophilic group provided by Vc. Vc improves the Vmax and Km of myrosinase on its
substrates [6].
Toxicants in Foods 311

2.3. Toxic Amino Acids

Toxic amino acids are rare amino acids that do not participate in protein synthesis, such
as homoserine, jenkolic acid, and 5-hydroxytryptophan. Toxic amino acids are usually the
structural analogs of protein amino acids and exert their toxicity as antimetabolites. Non-
proteinic amino acids are present in plants in quite low levels and certain non-proteinic amino
acids are found only in specific plants. For example, theanine is found only in the Camellia
family and the highest content is recorded in tea seeds, reaching 1% of dry weight; 5-
hydroxytryptophan is present only in the pea family and its content in Griffonic seeds
amounts to 14% by dry weight.
It should be noted that not all non-proteinic amino acids are toxic. Some rare amino
acids, such as theanine and alliin, can impart foods with special characteristics and healthy
functions. A summary of some unusual amino acids in food plants is given in Table 10-6.

2.3.1. Classification
Up to hundreds of non-proteinic amino acids have been identified in animals and plants
and most of them are the intermediate or final products of certain metabolisms in animals and
plants. Toxic amino acids can be divided into two categories according to their toxicity:

Table 10-6. Some toxic amino acids commonly occurring in plant foods and forages [7,8]

Amino acid Source


N-(3-hydroxypyridone-4)-2-aminopropionic acid Many species of Mimosoidae (a subfamily
(mimosine) of Leguminosae)
2-Amino-4-(guanidinooxy)butyric acid Many species of Papilionoidae (a
(canavanine) subfamily of Leguminosae
L-2-Amino-6-amidinohexanoic acid (indospicine) Tropical legume Indigofera spicata
5-Hydroxy-L-tryptophan (5-HTP) Griffonia simplicifolia
2-Amino-3-methylaminopropionic acid Cycas spp.
L-3,4-dihydroxyphenylalanine (L-DOPA) Vicia and some other legumes
3-Methylenecyclopropylpropionic acid
Plants of Sapindaeceae family
(hypoglycin A)
Seleno-amino acids (methylselenocysteine,
Plants grown on selenium-rich soils
selenocystathionine, selenocystine)
3-Cyanoalanine and 4-Glutamylcyanoalanine Vicia spp.
δ-Acetyl ornithine Lathyrus spp.
Homoserine and o-oxalylhomoserine Lathyrus spp.
Lathyrine, γ-methyl glutamic acid, γ-
Lathyrus spp.
hydroxynorvaline, and γ-hydroxyhomoarginine
Homoarginine and pipecolic acid Lathyrus and Vicia spp.

Bone poisoning-causing amino acids

β-aminopropionitrile and β-(N–γ-glutamyl)- aminopropionitrile are the representatives of


this category. Both the two amino acids exert their toxicity by inhibiting the formation of
cross linkages between the polypeptide chains during the synthesis of collagen and elastin [9].
312 Wang Dongfeng, Guoqing Huang and Shuhui Wang

Neurotoxic amino acids

The representatives are α, γ-diaminobutyric acid, β-cyano-L-alanine, β-N-oxalyl-α, β-


diamino-propionic acid and L-homoarginine.

2.3.2. Toxicity
Toxic amino acids are not essential for human body and can interfere with the normal
amino acid metabolism. Djenko sickness is a disorder of the urinary tract and most cases are
caused by the intake of djenko bean, which contains the toxic djenkolic acid. In some legume
species, the contents of djenkolic acid can reach 1~2% and its content in black varieties is as
high as 3~4%. Djenkolic acid shares a similar structure with cysteine and thus can interfere in
the metabolism of this essential amino acid.

2.4. Lectins

2.4.1. Classification
Lectins occur widely in plants, animals, and microorganisms. Exogenous lectins are also
known as vegetable haemagglutinin. They are sugar-binding proteins or glycoproteins of non-
immune origin and can lead to selective red blood cell agglutination in human blood. Lectins
can reversibly bind to specific monosaccharides or oligosaccharides. Most lectins contain
more than one sugar-binding site for cross connection with oligosaccharides. The molecular
weight of lectins ranges from 91 kDal to 130 kDal. Lectins are natural antigen of red blood
cells and are heat stable. Some lectins are highly toxic to experimental animals. For example,
when mouse are orally fed with 80 mg/kg garlic lectin for consecutively 7 days, both the
appetite and body weight are significantly reduced.
Vegetable hemagglutinin is firstly discovered in castor seeds. To present, a large number
of lectins with different properties have been identified. According to structure, lectins are
divided into merolectin, hololectin, chemerolectin, and superlectin. Based on the specificity to
sugars, lectins can be of the fucose type, the galactose/N-acetyl galactosamine type, the N-
aceytl glucosamine type, the mannose type, the sialic acid type, or the glycoconjugate type.
According to the origin, lectins can be divided into legume-derived lectins, mannose-binding
lectins, chitin-binding lectins, type-2 ribosome-inactivating proteins (RIP), and lectins derived
from other crops. According to the agglutination to red blood cells, lectins can be categorized
as specific and non-specific lectins. Based on evolution and structure correlation, lections can
be divided into 7 families. Table 10-7 lists the structure, sugar specificity, and distribution of
the 7 families in crops.
Based on reported researches, Peumans W J divided the lectins that have been identified
in crops into 5 categories: (1) legume lectins. Legume lectins are a family of structurally
related lectins that occur exclusively within the legume family. In Spite of their structural and
evolutionary relationships, legume lectins exhibit a wide range of carbohydrate-binding
specificities. (2) Monocot mannose-binding lectins. Monocot mannose-binding lectins occur
in at least five different families (Amaryllidaceae, Alliaceae, Araceae, Liliaceae and
Orchidaceae). They all have a similar molecular structure and sugar-binding specificity. (3)
chitin-binding lectins.
Toxicants in Foods 313

Table 10-7. Properties of the 7 families and their distribution in crops

Family Structure Specificity Distribution


Man/Glc Gal/Gal NAc
Leguminous (GlcNAc)n Fuc
Β-sandwich Leguminosae
lectin Siaα2,3 Gal/GalNAc
complex
Orchidaceae, Liliaceae,
Monocot
Amaryllidaceae, Araceae,
mannose-binding Β-barrel Man
Alliaceae, Bromeliaceae,
lectin
Iridaceae
Family Structure Specificity Distribution
Hevein domain- Hevein (GlcNac)n Gramineae, Solanaceae,
containing chitin- domain Urticaceae, Loranthaceae
binding lectin
Type-2 ribosome- β-trefoil Gal/GalNac Euphorbiaceae,
inactivating Siaα2,6 Gal/GalNAc Caprifoliaceae, Loranthaceae,
proteins Iridaceae, Ranunculaceae,
Lauraceae, Passifloraceae,
Liliaceae, Leguminosae
Cucurbitaceae Unknown (GlcNac)n Cucurbitaceae
Phloem Lectins
Breadfruit lectin Β-prism Gal/T-antigen Moraceae, Convolvulaceae,
Man Compositae, Musaceae,
Gramineae, Cruciferae
Amaranthaceae Β-trefoil GalNAc/T-antigen Amaranthaceae
lectin

Table 10-8. Legume lectins [10]

Species (common Tissue Content in Oral Heat Harmful lectin in food


name) raw material toxicity stability a Raw Processed
(g/kg)
Arachis hypogaea Seed 0.2-2 Slight Unstable Yes Yes
(peanut)
Glycine max Seed 0.2-2 Slight Low Yes No
(soybean)
Lens culinaris Seed 0.1-1 Slight Unstable Yes No
(lentil)
Phaseolus Seed 1-10 High Moderate Yes Possibly
coccineus (scarlet
runner bean)
Phaseolus lunatus Seed 1-10 High Moderate Yes Possibly
(lima bean)
Phaseolus Seed 1-10 High Moderate Yes Possibly
acutifolius (tepary
bean)
314 Wang Dongfeng, Guoqing Huang and Shuhui Wang

Table 10-8. (Continued)

Species (common Tissue Content in Oral Heat Harmful lectin in


name) raw material toxicity stability a food
(g/kg)
Phaseolus Seed 1-10 High Moderate Yes Possibly
vulgaris (kidney
bean)
Pisum sativum Seed 0.2-2 Minor Unstable Possible No
(garden pea)
Vicia faba (fava Seed 0.1-1 Minor Unstable Possible No
bean)
Note: a Heat stability (of purified lectin in aqueous solution); very high: withstands 90°C; high:
withstands 80°C; moderate: withstands 70°C; low: withstands 6OT; unstable: denatured at 60°C.
The definitions also apply to Table 10-6.

Chitin-binding lectins are found in five taxonomically unrelated families (Gramineae,


Solanaceae, Urticaceae, Papaveraceae and Amaranthaceae). Although there are some
differences in their overall molecular structure, all chitin-binding lectins are composed of
similar domains and exhibit comparable specificity. (4) A dozen type-2 ribosome-inactivating
proteins (RIP) have been isolated from plants belonging to taxonomically unrelated families.
All type-2 RIP are chimeric proteins that are composed of a catalytically active A chain that is
covalently linked through a disulphide bridge to a carbohydrate-binding chain. With the
exception of a lectin from elderberry, all type-2 RIPs exhibit specificity towards galactose or
N-acetylgalactosamine. (5) Other plant lectins [10].

Table 10-9. Mannose-binding lectins [10]

Species (common Content in raw Harmful lectin in food


Tissue Oral toxicity Heat stability
name) material (g/kg) Raw Processed
Allium ascalonicum
Bulb 0.01-0.1 Nontoxic Moderate No No
(shallot)
Allium cepa (onion) Bulb <0.01 Nontoxic Moderate No No
Allium porrum
Leaf <0.01 Nontoxic Moderate No No
(leek)
Allium sativum
Bulb 0.5-2 Nontoxic Moderate No No
(garlic)
Allium ursinum
Bulb 1-5 Nontoxic Moderate No No
(ramsons)
Colocasia esculenta
Tuber 1-5 ND Moderate No No
(taro)
Xanthosoma
Tuber 1-5 ND Moderate No No
sagittifolium (taro)

2.4.2. Contents and Properties


Based on reported researches, Peumans W J et al. summarize the contents, heat stability,
and harmfulness to human body of the 5 categories of lectins, as shown in Tables 10-8 to 10-
12.
It could be seen that among the legume lectins, runner bean, lima bean, tepary bean, and
kidney bean have higher lectin contents and the corresponding processed foods are possibly
toxic. Soybean foods that are improperly processed, such as by insufficient heating, may lead
Toxicants in Foods 315

to lectin poisoning. Experiments have proved that when mouse are fed lectin-containing crude
soybean powder, the mouse will suffer either enterocyte breakdown, leading to intestine
malfunction, or reduced activity of hydrolytic enzymes in stomach and intestine, leading to
decreased nutrient adsorption and inhibited growth.

Table 10-10. Chitin-binding lectin [10]

Species (common Tissue Content in Oral Heat Harmful lectin in food


name) raw material toxicity stability Raw Processed
(g/kg)
Hordeum vulgare Seed <0.01 ND High Yes Possible
(barley)
Oryza sativa (rice) Seed <0.01 ND High Yes Possible
Secale cereale Seed <0.01 ND High Yes Possible
(rye)
Triticum vulgare Seed <0.01 Moderate High Yes Yes
(wheat)
Triticum vulgare Germ 0.1-0.5 Moderate High Yes Yes
(wheat)
Amaranthus Seed 0.1 Nontoxic Very high Unkown Unkown
caudatus
(amaranth seeds)
antifungal protein
Cyphomandra Seed <0.01 ND Unkown Unkown Unkown
betacea (tamarillo)
Lycopersicon Fruit <0.01 Nontoxic High Possibly Possibly
esculentum
(tomato)
Solanum Tuber 0.01-0.05 No High Not for direct Possibly
tuberosum (potato) consumption

Table 10-11. Type-2 ribosome-inactivating proteins [10]

Content in Harmful lectin in food


Species (common Oral Heat
Tissue raw material
name) toxicity stability Raw Processed
(g/kg)
Not for
Ricinus communis direct
Seed 1-5 Lethal Unstable None
(castor seeds) consumpti
on
Sambucus nigra
Fruit 0.01 ND Moderate Yes Possibly
(elderberry)

2.5. Saponins

Saponins are a complex group of compounds and consist of saponin, sugar, uronic acid or
other organic acids. Most saponins are white and amorphous powders and tastes bitter and
spicy. Saponins are insoluble in nonpolar solvents but are soluble in water-containing polar
solvents. Saponins can irritate the mucous coat of digestive tract and lead to local hyperemia,
316 Wang Dongfeng, Guoqing Huang and Shuhui Wang

swelling, and hemorrhagic inflammation. The symptoms of saponin poisoning include


nausea, vomiting, diarrhea, abdominal pain.

Table 10-12. Other plant lectins [10]

Content in Harmful lectin in food


Species raw Oral Heat
Tissue
(common name) material toxicity stability Raw Processed
(g/kg)
Amaranthus
caudatus Seed 0.1-0.5 Nontoxic Unstable No No
(grain amaranth)
Artocarpus
integrifolia Seed 0.5-2 ND ND Unknown Unknown
(jact fruit)
Cucurbita pepo
Fruit <0.01 ND ND Possibly Unknown
(pumpkin)
Musa
paradisiac Fruit <0.01 ND ND Unknown Unknown
(banana)

Saponins are widely distributed in plants as well as in some marine animals. The reports
on saponins-induced food poisoning are emerging in recent years. For example, poisoning
caused by the ingestion of improperly cooked kidney bean is quite common in China and
saponins have been confirmed as a cause of the poisoning. One of the most intensively
studied saponins is tea saponins and their structure, composition, and physiological-
biochemical properties have been well elucidated. The following describes the structure,
properties, and toxicity of saponins by taking tea saponins as an example.

2.5.1. Structure of Tea Saponins


A saponin consists of ligand, glycoside, and organic acid. Chemically, a saponin has a
triterpene or steroid backbone linked to one or more sugar groups. There are thus two broad
subdivisions of saponins: those with triterpenoid aglycones and those with steroid aglycones.
Tea saponins contain one of the following four ligands: R1-barrigenol, theasapogenol B,
theasapogenol D, and A1-barrigenol. Of the four ligands, R1-barrigenol and A1-barrigenol
are present only in theafolisaponin and cannot be found in tea seed saponins. The structure
and properties of the saponins are illustrated in Figure 10-7 and Table 10-13. Both
theafolisaponin and tea seed saponins have triterpenoid aglycones as their ligands.

Table 10-13. Chemical properties of theafolisaponin

Molecular
Ligand Melting point (°C) Empirical formula
weight
R1-barrigenol 303~308 506 C30H50O6
Theasapogenol B 284~288 490 C30H50O5
Theasapogenol D 285~286 474 C30H50O4
A1-barrigenol 285~287 490 C30H50O5
Toxicants in Foods 317

R1-barrigenol Theasapogenol B

Theasapogenol D A1-barrigenol

Figure 10-7. Ligands of theafolisaponin.

Figure 10-8. Glycoside structure of tea saponins.

Theafolisaponins consist of sapogenin, glycoside, and organic acid. In addition to the


four ligands mentioned above, three more ligands have been found, but their structures
remain unclear.
The organic acid in tea seed saponins might be angelic acid, tiglic acid (E-2-methylbut-2-
enoic acid), or acetic acid. In theafolisaponins, the organic acid is one of angelic acid, tiglic
acid, or cinnamic acid. Arabinose, xylose, galactose, and glucuronic acid are major glycosides
contained in theafolisaponins and tea seed saponins (see Figure 10-8).

2.5.2. Properties and Toxicity of Tea Saponins


Tea saponins are colorless crystals with fine cylindrical structure. The saponins taste
bitter and spicy with strong foamability. Crystal tea saponins are insoluble in ether,
chloroform, benzene or other nonpolar solvents, less soluble in cold water, anhydrous
methanol or anhydrous ethanol, soluble in warm water, carbon disulfide, and ethyl acetate,
and readily soluble in water-containing ethanol, water-containing methanol, n-butanol, glacial
acetic acid, acetic anhydride, pyridine, and many other polar solvents. Tea saponins produce
green color after reacting with 5-methyl resorcinol hydrochlorate and their aqueous solutions
are acidic in the methyl red test.
318 Wang Dongfeng, Guoqing Huang and Shuhui Wang

The toxicity of saponins is often attributed to their hemolysis-inducing capability. Tea


saponins can damage the red blood cells of animals and result in hemolysis. The toxicity of
saponins is thus measured by the maximum valid dilution that still yields hemolysis. The
hemolytic activity of tea saponins is lower than Camellia sasanqua saponins, but comparable
to those derived from Camellia sinensis and Camellia japonica. Tea saponins exhibit toxicity
only to red blood cells (including the nucleated red cells of fish blood and chicken blood and
non-nucleated human blood) and are non-toxic to leucocytes. The hemolytic mechanism of
tea saponins is reported related to the permeability change of cell membrane. Tea saponins
can injury the cholesterol-containing cell membrane and lead to cytoplasm leakage and cell
disintegration. The contact between saponins and blood is the prerequisite of hemolysis.
Hence, oral ingestion of saponins does not cause damage in animals or human body.
Tea saponins are highly toxic to cold-blood animals, such as fish, frog, and blood sucker,
even in low concentrations, but are non-toxic to higher animals when orally administered.
Camellia sasanqua saponins are the most toxic to fishes, followed by those derived from
Camellia japonica and Camellia sinensis in sequence. Their LD50s are 0.25 mg/L, 3.8 mg/L,
and 4.5 mg/L respectively. High water salinity improves the toxicity, as evidenced by the
LD50 of 5 mg/L for freshwater fishes and that of less than 1 mg/L for seawater fishes.
Osmotic pressure also affects tea saponin toxicity. In the same tea saponins concentration,
fishes display low mortality in the salinity range 4‰~10‰, but a further increase or decrease
of the salinity accelerates fish death. Besides, the toxicity to fish increases with elevated
temperature. Tea saponins can be hydrolyzed in alkaline conditions and thus inactivated.
Since seawater is slightly alkaline, tea saponins are degraded within 48 h and do not cause the
environmental pollution concern.
Tea saponins injury gills and induce hemolysis in fishes. Tea saponins have no adverse
effect on shrimps, which also use gill for respiration. There are two reasons for the selective
toxicity. The shrimp gill is a cuticle layer zone that develops from the cuticle layer. The
epidermis of shrimp gill consists mainly of chitin and protein, which differs significantly
from that of fish gill in structure and composition. Besides, the oxygen carrier in fish blood is
heme with Fe2+ as its core. In contrast, the oxygen carrier in shrimp blood is hemocyanin and
its core is Cu2+. The specific toxicity of tea saponins to fishes has been utilized in the
aquaculture industry. Tea saponins have been used as pond-clearing agent of shrimp rearing
ponds for removing harmful fishes. Tea saponins have been successfully in hundreds of
hectares of sea prawn rearing ponds along the coastlines of the East China Sea, the Yellow
Sea, and the Bohai Sea.

2.6. Toxins in Aquatic Organisms

2.6.1. Tetrodotoxin (TTX)


TTX is a neurotoxin found in a variety of fish species. Pure TTX is odorless and colorless
crystals and its formula is C11H17O8N3, corresponding to the molecular weight of 319.27.
TTX is insoluble in water or organic acids, but soluble in weakly acidic aqueous solutions.
TTX is unstable in strong acid and alkaline solutions, but is resistant to UV and sunlight.
Exposure to UV for 48 h or to sunlight for one year does not change the toxicity of TTX.
TTX is not degraded by pepsin, trpsin or amylase and is stable against salts. TTX is heat
stable in neutral and faintly acidic conditions. This is why cooked globefishes still cause TTX
poisoning.
Toxicants in Foods 319

TTX is an extremely toxic natural toxin and is roughly 100 times more poisonous than
potassium cyanide. The LD50 (i.v.) to mice is 8.7μg/kg.

2.6.2. Paralyfric Shellfish Poisons


Paralyfric shellfish poisons (PSPs) are the derivatives of tetrahydropurine. These
compounds are produced by Gonyaulax catenella and other dinoflagellate species, and the
algal toxins (phycotoxins) contaminate shellfish during the filter-feeding of bivalve molluscs.
To present, more than 20 PSPs have been identified. Based on structure, PSPs can be divided
into carbamoyl type, such as saxiltoxins (STX), neosaxitoxins (neoSTX) and gonyautoxins
(GTX 1-4); N-sulfocarbamoyl type, such as C1-4, GTX5, and GTX6; and decarbomyl type,
such as dcSTX and doGTX2-3. PSPs are soluble in water, methanol, and ethanol and are
stable against acid and heat treatment.
PSPs are nerve and muscle paralyzants and take effect by disrupting the Na/K channel.
Symptoms of PSP poisoning include nausea, vomiting, diarrhea, abdominal pain, and tingling
or burning of lips, gums, tongue, face, neck, arms, legs, and toes.

2.6.3. Ciguatera Toxins


Ciguatera is a foodborne illness caused by eating reef fishes whose flesh is contaminated
with toxins originally produced by dinoflagellates such as Gambierdiscus toxicus. The
poisons causing ciguatera are termed ciguatera toxins and are composed of cigatoxins
(CTXs), maitotxin (MTX), and scaritoxin (STX), of which, CTX and MTX are the major
constituents.
CTXs consist of 13 fused ether rings and are colorless, heat resistant, and amorphous.
CTXs are soluble in polar organic solvents, such as methanol, ethanol, and acetone, but
insoluble in benzene or water. CTXs are susceptible to oxidation. Its LD50 (i.v.) to mice is
0.45 μg/kg and is 20 times poisonous than TTX.
MTX is a polyether and contains up to 31 ether rings. It is ahighly polar molecule and is
soluble in water, methanol, ethanol, and dimethylsulfoxide but insoluble in chloroform,
acetone or acetonitrile. Pure MTX is white crystal and is extremely susceptible to oxidation.
Incubation with 1mol/L hydrochloric acid or ammonium hydroxide solution with heating
does not change MTX toxicity. Its LD50 (i.v.) to mice is 0.15μg/kg, which is 2 times more
poisonous than CTXs.
STX is fat soluble and shares similar chemical and chromatographic properties with
CTXs, but differ in polarity as evidenced by TLC and DEAE chromatograph analysis. STX
has no absorbance in wavelengths above 220nm and its structure is still not elucidated to the
present.

2.6.4. Diarrhetic Shellfish Poisons


Diarrhetic shellfish poisons (DSPs) are fat soluble polyether compounds. Based on their
backbone structures, DSPs are divided into two groups:
OA-group toxins: This group includes okadaic acid (OA) and its derivatives
dinophysistoxin-1 (DTX-1) and dinophysistoxin-2. OA is a derivative of C38 fatty acid and
its formula is C44H68O13. The structures of OA, DTX-1, and DTX-2 are given in Figure 10-9.
Toxins of this group are lipophilic and heat stable. They are produced by dinoflagellates and
can be found in various species of shellfish, mainly in filter-feeding bivalve molluscs such as
oysters, mussels, scallops, and clams.
320 Wang Dongfeng, Guoqing Huang and Shuhui Wang

PTX-group toxins: Pectenotoxins (PTXs) are a group of polyether-lactone toxins. To


date, more than 15 different analogues have been isolated and characterized. PTX-group
toxins are exclusively produced by Dinophysis species. They can be found in filter-feeding
bivalve molluscs such as oysters and mussels. PTX-group toxins are heat stable, but they are
easily destroyed in strong alkaline and acidic solutions.
The characteristic symptoms of diarrhetic shellfish poisoning include diarrhoea, nausea,
vomiting and abdominal pain. The LD50 of OA to rat is 160μg/kg. In additoin to causing
diarrhea, OA is strongly cancerigenic. PTXs are much more toxic (with LD50 in the range 16-
77μg/kg) and can cause liver damage.

2.6.5. Actinotoxins
Actinotoxins are toxin derived from extracts of the tentacles of sea anemones. To present,
more than 60 kinds of cytolysin toxins have been isolated from sea anemones. Cytolysins are
substances or antibodies raised by microorganisms, plants or animals that are specifically
toxic to individual cells. The molecular weights of sea anemone-derived cytolysins range
from 15000 to 20000 and can selectively bind to the lipids on cell membranes, causing pain,
inflammation, and muscular paralysis. The most intensively studied actinotoxin is the
cytolysin isolated from anthoplerin. The molecular weight of the cytolysin is 17000 and is
characterized by a long hydrophobic β-sheet section and 5 short hydrophobic β-hydrophobic
sections in the N terminal, in which, 60%-70% of amino acids are hydrogen bonded and form
a transmembrane structure. The C terminal is highly hydrophilic and locates on the outside
surface of membrane, which forms a transmembrane channel.

Figure 10-9. Structures of OA and DTX-1, and DTX-2. OA: R1=CH3, R2=H, R3=H, DTX-1: R1=CH3,
R2=CH3, R3=H, DTX-2: R1=H, R2=CH3, R3=H.

2.6.6. Conotoxins
Conotoxins (CTXs) are a group of neurotoxic peptides isolated from the venom of the
marine cone snail, genus Conus. CTXs are highly poisonous to human body and the
selectivity varies with the living habits of cone snails. Among the CTXs identified, that
isolated from Conus gegraphus Linnaeus is the most poisonous.
The structures of more than 100 CTXs have been elucidated. A majority of CTXs
characterized from venoms so far are 12–30 amino acids (AA) in length and contain one or
more disulfide bonds. CTXs are initially translated as propeptide precursors, approximately
80–100 AA and the biologically active toxins are produced by proteolytic cleavage from the
precursors. Conotoxins can act on a wide range of targets and are highly tissue specific. CTXs
have been used as probes in the classification and identification of ion channels and receptors.
CTXs are potential drugs or leading compounds for new drug development.
Toxicants in Foods 321

2.6.7. Cyanotoxins
Cyanotoxins are produced by cyanobacteria (also known as blue-green algae). The
mechanisms of cyanobacterial toxicity currently described and understood are very diverse
and range from hepatotoxic, neurotoxic and dermatotoxic effects to general inhibition of
protein synthesis. Cyanotoxins fall into three broad groups of chemical structure: cyclic
peptides (including microcystins and nodularins), alkaloids (including neurotoxic anatoxins
and saxitoxins, cytotoxic cylindrospermopsin and demethoxy-cylindrospermopsin, and
dermatotoxic aplysiatoxins and lyngbyatoxin) and lipopolysaccharides.

3. EXOGENOUS TOXICANTS IN FOODS


During the processing, package, storage, and transport of foods, non-intrinsic components
are inevitably added or generated. Besides, plants and animals might accumulate hazardous
compounds for the environment and some of which might be absorbed by consumers and
pose potential risks. The intrinsic components and various additives that are supplemented in
compliance with regulations impart foods with nutrition and flavor. However, the
contaminants produced during food processing, package, storage, and transport or due to
environmental pollution are unnecessary for foods and most of the contaminants might cause
potential risk on food safety.

3.1. Heavy Metals

The nutritional status of plants and animals are closely associated with the environment
and fertilizer or feed. When minerals in the environment and fertilizer or feed are deficient or
excessive, the minerals in the corresponding foods are also in shortage or excess. Plants and
animals accumulate both essential and harmful minerals and finally affect the mineral
contents of top predators in the food chain. For example, the Keshan disease and kashin-beck
disease suffered by residents in Se-deficient areas are possibly due to Se deficiency in foods.
If the human body intakes excessive minerals, especially heavy metals, poisoning occurs. The
world-famous Minamata disease is a result of Hg pollution.
In addition to water and air, the contents of heavy metals in human body are mainly
affected by their contents in foods. When human beings consume foods derived from plants
or animals with strong heavy metal accumulation capabilities, the metals will exceed
thresholds and cause potential harm on human body.

3.1.1. Content and Toxicity of Heavy Metals


All metals are toxic when their contents exceed certain thresholds. The deficiency of
essential metals can lead to growth retardation, declined propagation, and even death. In the
early stage of essential metal deficiency, the growth and propagation can be recovered by
supplementing the corresponding elements.
The toxicity of metals depend on their contents and valence and the presence of other
metals as well. For example, Cu increases the toxicity of Hg but reduces the toxicity of Mo.
In turn, Mo significantly reduces Cu adsorption and causes Cu deficiency. Cd interferes in the
adsorption of Cu and Cu in low valence reduces the tolerance to Cd toxicity.
322 Wang Dongfeng, Guoqing Huang and Shuhui Wang

Living bodies do not show deficiency for non-essential metals, such as Be, Hg, Pb, and
Sb. However, once the contents of these metals exceed certain thresholds, toxicity symptoms
appear.

3.1.2. Poisoning Mechanism


The poisoning mechanisms of metals are very complex. In addition to their contents in
human body, the toxicity of metals is also affected by their ingestion pathway, solubility,
presence form, physic-chemical properties, and the status of human body. Generally, metals
exert their toxicity through one or more of the following mechanisms:

1. Heavy metals disrupt the functional groups of bioactive molecules. For example,
Hg(II) and Ag (II) can bind to the –SH of cysteine residues in enzymes and interrupt the
enzymatic reactions mediated by the –SH group.
2. Heavy metals replace the metal cofactors in biomolecules. The activity of
metalloenzymes is closely associated with metals. Due to the different macromolecule-
binding stability of metals, a metal with a higher stability constant replaces another metal
with lower stability constant and consequently inactivate the metalloenzyme.
3. Heavy metals change the conformation or higher-order structures of biomolecules. For
example, polynucleotides are units for genetic information storage and transfer. Heavy metals
can bind to the units and change their structures. This is why heavy metals readily cause
cancer and teratogenesis.

Proteins, phosphatides, some sugars and nucleonic acids can provide atoms for
coordinating with metal ions, such as the nitrogen atom in imidazole (histidine), NH2 (lysine),
purine and pyrimidine bases in DNA and RNA, oxygen atom in the hydroxyl group (serine,
tyrosine), COO- (glutamic acid and aspartic acid), PO43- (phospholipids, nucleic acids), and
sulfur atom in –SH (cystine) and SR (methionine, CoA). The three atoms are good ligands for
metals.
In metal activated enzymes, the prosthetic metals often bind loosely to the enzymes. The
metals can be easily replaced by other metals and the functions of the enzymes are destroyed
as a result. In metalloenzymes, metals bind tightly to the enzymes and their replacement by
other metals is difficult. However, during enzyme synthesis, metal replacement may occur
readily.
Cell wall, cell membrane, and organelle membrane are the main barrier of metal intake.
Generally, lipophilic metal ions with less charge can permeate the barriers easier. For
example, CH3Hg+ has a higher permeability than Hg2+ and the permeability of (CH3)2Hg is
greater than CH3Hg+. Based on these facts, it can be deduced that M2+ coordinated with a
organic ligand can enter cells easier than its hydrate form [M2+(H2O)n].
Metals can change the permeability of cellular membranes by initiating lipid peroxidation
and the changes vary with the affinity of metals to the ligands on membrane surface. The
peroxidative degradation of lipids in cell and organelle membranes is lethal to lif