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Food Emulsifiers and
Their Applications
Edited by
Gerard L. Hasenhuettl
Kraft Foods
Richard W. Hartel
University of Wisconsin
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12345678910 XXX 0100999897
Library of Congress Cataloging-in-Publication Data
Food emulsifiers and their applications/edited by Gerard L.
Hasenhuettl and Richard W. Hartel.
p. em.
Includes bibliographical references and index.
ISBN 978-1-4757-2664-0 ISBN 978-1-4757-2662-6 (eBook)
DOI 10.1007/978-1-4757-2662-6
1. Food additives. 2. Dispersing agents. I. Hassenhuettl, Gerard L., 1944-
II. Hartel, Richard W., 1951-
TP455.F667 1997
664'.06-dc21 97-1647
CIP
Contributors xiii
Preface XV
ONE
Overview of Food Emulsifiers 1
Gerard L. Hasenhuettl
1.1 Introduction 1
1.2 Emulsifiers as Food Additives 2
1.3 Emulsifier Structure 5
1.4 Emulsifier Functionality 7
References 9
TWO
Synthesis and Composition
of Food-Grade Emulsifiers 11
R. J. Zielinski
2.1 Introduction 11
2.2 Mono- and Diglycerides 13
2.3 Propylene Glycol Monoesters 15
2.4 Lactylated Esters 18
2.5 Polyglycerol Esters 23
vii
viii Food Emulsifiers and Their Applications
THREE
Analysis of Food Emulsifiers 39
Gerard L. Hasenhuettl
3.1 Introduction 39
3.2 Thin-Layer and Column Chromatography 40
3.3 Wet Chemical Analysis 43
3.4 Physical Methods 52
3.5 Instrumental Methods 54
3.6 Setting Specifications for Food Emulsifiers 62
References 63
FOUR
Carbohydrate/Emulsifier Interactions 67
Lynn B. Deffenbaugh
4.1 Introduction 67
4.2 Inclusion Complexes of Starch 68
4.3 Effects of External Lipid Materials on
Starch Properties 69
4.4 Factors Affecting Complex Formation 81
4.5 Physical Properties of Starch/Emulsifier Complexes 85
4.6 Summary 90
References 90
Contents ix
FIVE
Protein/Emulsifier Interactions 95
Martin Bos, Tommy Nylander, Thomas Arnebrant,
and David C. Clark
5.1 Introduction 95
5.2 Protein Stability 97
5.3 Protein/Surfactant Interactions 98
5.4 Protein/Phospholipid Interactions 120
5.5 Protein/Emulsifier Interactions-Food Applications 132
References 137
SIX
Physicochemical Aspects of an Emulsifier
Functionality 147
Bjorn Bergenstahl
6.1 Introduction 147
6.2 Surface Activity 148
6.3 Solution Properties of Emulfiers 149
6.4 The Use of Phase Diagrams to
Understand Emulsifier Properties 153
6.5 Examples of the Relation between
Phase Diagrams and Emulsion Stability 154
6.6 Some Ways to Classify Emulsifiers 161
6.7 The Emulsifier Surface 167
References 171
SEVEN
EIGHT
Applications of Emulsifiers
in Baked Foods 211
Frank T. Orthoefer
8.1 Introduction 211
8.2 History of Emulsified Shortenings 211
8.3 Emulsifier Function in Baked Goods 213
8.4 Role of the Shortening 215
8.5 Role of the Emulsifier 216
8.6 Emulsifier Interaction with Bakery Components 221
8. 7 Applications in Baked Goods 225
8.8 Summary 233
References 234
NINE
Emulsifiers in Confectionery 235
Mark Weyland
9.1 Introduction 235
9.2 Emulsifiers in Chocolate and Compound Coatings 236
9.3 Antibloom Agents in Chocolate and
Compound Coatings 244
9.4 Other Emulsifiers Used in Coatings 248
9.5 Emulsifiers in Nonchocolate Confectionery 249
Contents xi
TEN
Margarines and Spreads 255
Eric Flack
10.1 Introduction 255
10.2 Early Development 256
10.3 Yellow-Fat Consumption 257
10.4 Definitions and Descriptions 258
10.5 Structure and Raw Materials 262
10.6 Fat Crystallization 263
10.7 Emulsifiers 267
10.8 Processing 270
10.9 Reduced- and Low-Fat Spreads 274
10.10 Oil-in-Water Spreads 278
10.11 Liquid Magarine 278
10.12 Summary 279
References 279
ELEVEN
Emulsifier Trends for the Future 281
Gerard L. Hasenhuettl
11.1 Globalization in the Food Industry 282
11.2 Nutritionally Driven Changes in Food 282
11.3 Trends toward Safer Emulsifers 283
11.4 Emulsifer Structure and Interactions
with Other Ingredients 284
11.5 Enzymatic Synthesis of Food Emulsifiers 285
References 286
Index 287
Contributors
Thomas Arnebrant
Department of Pharmaceutical Analysis
Ferring AB
Box 30047
S-216 13 Malmo, Sweden
Bjorn Bergenstahl
Institute for Surface Chemistry
P.O. Box 5607
S-114 86 Stockholm, Sweden
Martin Bos
TNO Nutrition & Food Research Institute
P.O. Box 360
3700 AJ Zeist, The Netherlands
David C. Clark
DMV International
NBC-Iaan 80
P.O. Box 13
5460 BA Veghal, The Netherlands
xiii
xiv Contributors
Lynn B. Deffenbaugh
Hill's Pet Nutrition
Science & Technology Center
P.O. Bo.c 1658
Topeka, KS 66601-1658
Stephen R. Euston
New Zealand Dairy Research Institute
Private Bag 11029
Palmerston North, New Zealand
Eric Flack
Danisco Ingredients, Ltd.
North Way
Bury St. Edmonds
Suffolk, 1P32 6NP, United Kingdom
Gerard L. Hasenhuettl
Kraft Foods, Inc.
801 Waukegan Rd.
Glenview, IL 60025
Tommy Nylander
University of Lund, Chemical Center
Department of Physical Chemistry
P.O. Box 124
S-22100 Lund, Sweden
Frank T. Orthoefer
2004 Me Cracken St.
Stuttgart, AR 72160
Mark Weyland
Quest International
5115 Sedge Blvd.
Hoffman Estates, IL 60192
Richard J. Zielinski
Quest International
Joliet Technical Center
24708 Durke Dr.
Joliet, IL 60410-5249
Preface
Food emulsions have existed since long before people began to process foods for
distribution and consumption. Milk, for example, is a natural emulsion/colloid in
which a nutritional fat is stabilized by a milk-fat-globule membrane. Early
processed foods were developed when people began to explore the art of cuisine.
Butter and gravies were early foods used to enhance flavors and aid in cooking.
By contrast, food emulsifiers have only recently been recognized for their abil-
ity to stabilize foods during processing and distribution. As economies of scale
emerged, pressures for higher quality and extension of shelf life prodded the de-
velopment of food emulsifiers and their adjunct technologies. Natural emulsifiers,
such as egg and milk proteins and phospholipids, were the first to be generally
utilized. Development of technologies for processing oils, such as refining,
bleaching, and hydrogenation, led to the design of synthetic food emulsifiers.
Formulation of food emulsions has, until recently, been practiced more as an
art than a science. The complexity offood systems has been the barrier to funda-
mental understanding. Scientists have long studied emulsions using pure water,
hydrocarbon, and surfactant, but food systems, by contrast, are typically a com-
plex mixture of carbohydrate, lipid, protein, salts, and acid. Other surface-active
ingredients, such as proteins and phospholipids, can demonstrate either syner-
XV
xvi Preface
Acknowledgments
The editors express their apprec1atwn to Barbara Bagnuolo and Julie
Hasenhuettl for typing and proofreading large portions of the manuscript.
Gerard L. Hasenhuettl
Richard W. Hartel
Food Emulsifiers and
Their Applications
ONE
1.1 Introduction
Food emulsions, colloids, and foams have their origins in the evolution of ani-
mal species. Milk has a naturally occurring membrane that allows the disper-
sion of fat droplets into an aqueous environment. Early food formulations to
produce butter, whipped cream, cheese, and ice cream built upon the natural
emulsifiers present in the system. The development of mayonnaise in France
as a cold sauce utilized the natural egg phospholipids to disperse a liquid oil
into an acidified aqueous phase. The emulsifying power is still very impressive
by today's standards since it allowed up to 80% oil to be dispersed without in-
version to an oil-continuous emulsion. The invention of margarine by the
French chemist Hippolyte Mege-Mouries in 1889 utilized the solid fat of tal-
low to produce a stable oil-continuous emulsion to serve as a low-cost substi-
tute for butter. In this case, the emulsion had to be stable temporarily only
until the product was chilled.
Synthetic emulsifiers have come into common use only in the latter half of
the twentieth century. Their development was driven by the emergence of the
processed food industry, which needed technology to produce and preserve
quality products that could be distributed through mass market channels. A
key technical hurdle was to maintain product stability over an extended shelf
1
2 Food Emulsifiers and Their Applications
life. With small amounts of emulsifier, for example, salad dressings can be
stored on a shelf for more than a year without visible separation. Now, other
factors such as the development of rancidity limit consumer acceptance of ag-
ing products.
Detailed knowledge of the physical chemistry of emulsions is best obtained
when pure oil, water, and surfactants are used. Food emulsions, by contrast, are
extraordinarily complex systems. Commercial fats and oils are complex mixtures
of triglycerides that also contain small amounts of highly surface-active materi-
als (Friberg and Larsson, 1990). Salt content and pH in food emulsions vary
widely enough to have significant effects on their stability. Natural and commer-
cial emulsifiers are often complex mixtures that vary in composition between dif-
ferent manufacturers. Other food ingredients, such as proteins and particulates,
contribute surface activity that may dramatically alter the character of the emul-
sion. Because of all these complex relationships, the formulation of food emul-
sions grew up as an art, dominated by individuals having a great deal of
experience. The gradual development of sophisticated techniques such as elec-
tron microscopy, nuclear magnetic resonance, and chromatography/mass spec-
trometry has solidified the art with a scientific dimension.
The subject of food emulsions has been extensively reviewed by Friberg
and Larsson (1990), Krog et al. (1985), and Jaynes (1985). This book will be
oriented to the design, manufacture, and use of food emulsifiers as ingredients
to improve the quality and economy of processed foods.
2ICFR Typical
Emulsifier No. Functionalities applications
As with any other totally new food additive, the need to prove safety of the
product in foods at high levels of consumption requires extensive toxicity studies
and enormous documentation. The consequent financial and time commitments
make development of totally new emulsifiers unattractive for emulsifier manu-
facturers. A somewhat easier development approach is to petition for expanded
use (new applications or higher permitted levels) of emulsifiers that are already
approved. However, even this tactic may require several years of review.
In addition to national regulations, many food processors require their in-
gredients, including food emulsifiers, to be kosher so that their products are
acceptable to Jewish and many Islamic consumers. For emulsifiers to be
kosher, they must be produced from kosher-certified raw materials. This re-
quirement precludes the use of almost all animal fats. This is not much of a
problem since emulsifiers are easily produced from vegetable fats that can be
blended to give similar fatty acid compositions. The major concern in kosher
certification is to determine in advance whether the customer's rabbinical
council recognizes the Hekhsher (kosher symbol) of the producer's rabbi.
Products labeled as "all natural" must contain ingredients that have not
been chemically processed or modified. Only lecithin or other naturally occur-
ring materials such as proteins would be acceptable for these products.
-OH
Saturated; palmitate or stearate
H H
Unsaturated; oleate
Unsaturated; linoleate
also contain measurable concentrations of trans (E) fatty acids that have higher
melting points than the cis (Z) fatty acids.
Polar head groups may be present in a variety of functional groups. They
may be incorporated to produce anionic, cationic, amphoteric, or nonionic sur-
factants. Mono- and diglycerides, which contain an -OH functional group, are
the most widely used nonionic emulsifiers. Lecithin, whose head group is a
mixture of phosphatides, may be visualized as amphoteric or cationic, depend-
ing on the pH of the product.
Proteins may also be surface active due to the occurrence of lipophilic
amino acids such as phenylalanine, leucine, and isoleucine. lnterfacially ac-
tive proteins will fold so that lipophilic groups penetrate into the oil droplet
while hydrophilic portions of the chain extend into the aqueous phase.
Proteins in this configuration may produce a looped structure that provides
steric hindrance to oil-droplet flocculation and coalescence. Charged proteins
may also stabilize emulsions due to repulsion of like charged droplets.
Proteins may also destabilize water-in-oil emulsions, such as reduced fat mar-
garines, by causing the emulsion to invert.
Food emulsifiers may be thought of as designer molecules because the
structure and number of heads and tails may be independently varied. A very
useful conceptual tool is hydrophile/lipophile balance (HLB). The topic has
been extensively reviewed by Becher, so only a brief description will be pre-
sented here. The number and relative polarity of polar groups in a surface-ac-
Overview of Food Emulsifiers 1
tive molecule determine whether the molecule will be water- or oil-soluble (or
-dispersible). This concept has been quantitated by calculation of an HLB
value to describe a given emulsifier. High-HLB values are associated with
easy water dispersibility. Since conventional practice is to disperse the surfac-
tant into the continuous phase, high-HLB emulsifiers are useful for preparing
and stabilizing oil-in-water (0/W) emulsions. Low-HLB emulsifiers are useful
for formulation of water-in-oil (W/0) emulsions, such as margarine. Extreme
high or low values are not functional as emulsifiers since almost all of the mol-
ecule will be solubilized in the continuous phase. They would, however, be
very useful for full solubilization of another ingredient, such as a flavor oil or
vitamin, in the continuous phase. At some intermediate values of HLB, the
molecule may not be stable in either phase and will result in high concentra-
tion at the interface.
actions are not confounded. Robust design is also highly recommended so that
the food product has minimal sensitivity to uncontrolled noise factors.
The objective of this book is to provide the food industry professional or inter-
ested technical professional with an overview of what emulsifiers are, how they
are prepared, and how they are utilized in food products. Although in many
senses food emulsifiers have become commodity ingredients, sophisticated un-
derstanding and application in processed foods is likely to continue to advance.
References
Friberg, S., Larsson, K. (eds.) (1990). Food Emulsions, Marcel Dekker, New York.
Jaynes, E. {1985). "Applications in the food industry, II," in Encyclopedia of Emulsion
Technology {ed. P. Becher), Vol. 2, Marcel Dekker, New York, 1985, pp. 367--85.
Krog, N., Rilson, T.H., Larsson, K. (1985). "Applications in the food industry, " in
Encyclopedia of Emulsion Technology (ed. P. Becher), Vol. 2, Marcel Dekker, New
York, pp. 321-66.
TWO
2.1 Introduction
The food-grade emulsifiers are generally esters composed of a hydrophilic (wa-
ter-loving) end and a lipophilic (fat-loving) end. In general, the lipophilic end
is composed of stearic, palmitic, oleic, or linoleic acid or combinations of these
fatty acids. The hydrophilic end is generally composed of hydroxyl or carboxyl
11
12 Food Emulsifiers and Their Applications
groups. The melting point of the various esters within each family will be de-
termined by the melting point of the fatty acids used to prepare the emulsifier.
The melting points of the common fatty acids are given in Table 2.1. When
stearic and palmitic acids dominate, the ester will be solid and relatively high-
melting; when oleic and linoleic acids dominate, the ester will be low-melting
and could be a liquid at room temperature. The fatty acids present in an emul-
sifier may be obtained from either a fat or oil or a fatty acid source. All fats and
oils are triglycerides, and the fatty acids can be obtained from the triglycerides
by a hydrolytic process followed by fractional distillation. Generally, either
natural or fully hydrogenated fats and oils are split to obtain the fatty acids.
This process is illustrated in Equation 1.
0
II
CH 2-0-C-R CH 2-QH + R--cOOH
I 0
II I
CH-o--c-R1 + 3H 20 CH-QH + R1--cOOH
I I
CH 2-0-C-R CH 2-0H + R2--cooH
I
0
Kosher oleic acid may be obtained from high-oleic safflower and high-oleic
sunflower seed oils. It may also be obtained from a specially purified tall oil
from pine trees. Commercial stearic acid may be of three types: (i) a mixture of
about 90% stearic acid and 10% palmitic acid, (ii) a mixture of about 70%
stearic acid, 30% palmitic acid, or (iii) a mixture of about 50% palmitic acid
and 50% stearic acid; all are known as stearic acid. Generally, partially hydro-
genated fats and oils are used to prepare plastic-type emulsifiers.
0 0 0
I II II
CH 2-0H CH-C-R CH 2-0-C-R CH2-0-C-R
0 0
I I I II I II
CH-OH CH-OH CH-0-C-R CH-0-C-R
0
I I I I II
CH 2-0H CH-0-H CH 2-0H CH2-0-C-R
Figure 2.1
14 Food Emulsifiers and Their Applications
0
II
CH 2-0H CH 2-0-c-R CH 2-0H CH-OH
I I 0
II heat I I
CH-QH + CH-0-C-R CH-OH + CH-QH
catalyst
0
I I 0
II I I II
CH 2-0H CH 2-0-C-R CH 2-0H CH-Q-C-R
0
II
CH 2-0H CH 2-0H CH 2-D-c-R
0 0
I heat I II I II
CH-OH + R-cOOH
catalyst
CH-O-c-R + CH-0-C-R
I I
CH 2-0-C-R
0
II I
CH 2-D-C-R
0
II
(2)
The catalysts commonly used in these processes are sodium hydroxide or hy-
drated lime (calcium hydroxide); the temperatures involved range from 200 to
250°C. The proportions of each of the products-free glycerine, monoglyceride,
diglyceride, and triglyceride-are purely dependent on the molar ratio of glycer-
ine and oil or glycerine and fatty acid used. The overall composition can be ap-
proximated using a random distribution of the free hydroxyl group and fatty acid
groups (Feuge and Bailey, 1946). The various fatty acid groups from the oil or
fatty acid are also assumed to he randomly distributed in the final product.
Once the desired ratio of oil or fatty acid and glycerine has been chosen to
yield the desired monoglyceride content, a number of different types of mono-
glyceride compositions are still potentially available. For example, if a 40%
monoglyceride composition is desired, then (i) the catalyst may be neutralized,
Synthesis and Composition of Food-Grade Emulsifiers 15
generally by phosphoric acid; (ii) the catalyst may be not neutralized; (iii) the
reaction mixture may be cooled to a determined temperature and the glycerine
that is insoluble in the final composition removed by decantation type process;
and (iv) the excess free glycerine may be removed from the reaction products
by vacuum distillation. A 40% monoglyceride prepared by a decantation-type
process (iii) may contain about 4% free glycerine, while a 40% monoglyceride
prepared by a vacuum-stripping process will typically contain less than 1%
free glycerine. A stripped 40% monoglyceride will typically be composed of
about 46% monoglyceride, 43% diglyceride, 10% triglyceride, and 1% glycer-
ine. Similarly, a 50% monoglyceride may be prepared by decantation from in-
soluble glycerine or by a vacuum-stripping process. A vacuum-stripped 50%
monoglyceride will typically contain about 55% monoglyceride, 38% diglyc-
eride, 5% triglyceride, and 2% free glycerine.
Under the conditions of high temperature, very low pressure, and an ex-
tremely short path, monoglycerides may be distilled and thus concentrated and
purified. This process is known as "molecular distillation." Typically, a 40%
monoglyceride mixture is subjected to molecular distillation to yield mono-
glycerides of over 90% purity. The nondistilled portion is recycled for an addi-
tional interesterification process to yield another 40% monoglyceride-type
composition for use as the feedstock.
As previously mentioned, either nonhydrogenated or partially hydrogenated
triglycerides, or saturated or unsaturated fatty acids, may be used to prepare
solid, plastic, or liquid monoglyceride mixtures.
The mono- and diglycerides have a generally recognized as safe (GRAS)
FDA status.
0
II
CH 2-QH CH 2-0-c-R
I I ~ heat
CH-OH + CH-O-C-R
catalyst
I I 0
II
CH 3 CH 2-0-c-R
Propylene Triglyceride
glycol
0 0
II II
CH 2-QH CH 2-o-c-R CH 2-0-c-R CH2-0H
I
+
I
+
I ~ I
CH-QH CH-OH CH-0-C-R + CH-OH +
I I I I
CH 3 CH 3 CH 3 CH2-QH
0 0 0
II II I
CH 2-0-c-R CH 2-Q-C-R CH 2-o-c-R
I I 0
I I 0
II
CH-QH + CH-0-C-R + CH-o-c-R
I I I 0
I
CH-QH CH 2-QH CH 2-o-c-R
0
I
CH 2-0H CH 2--o-c-R
I heat I
CH-OH + R-cOOH CH-OH +
catalyst
I I
CH 2 CH 3
0
II
CH 2-0-c-R
I
CH 2-0-C-R
~
I
(4)
tripropylene glycol monoesters. These esters are not allowed by the FDA as di-
rect food additives.
The FDA regulations for propylene glycol monoesters allow the use of all
edible oils and edible fatty acids. However, the majority of the propylene gly-
col monoesters commercially available contain very high percentages of
palmitic acid and stearic acid that are saturated fatty acids. Few unsaturated
propylene glycol monoesters are available at this time.
/R-COOH
OH f--- R-COOH OH
CH3
I I 0II I
CH 3-c-COOH f--- R-QH CH 3-C-c-o-cOOH f--- R-OH
I I
H H
Figure2.2
2.4.1.1 Lactylic &ters of Fatty Acids. Two types of lactylic esters of fatty acids
are commercially available. One composition contains approximately 60%
monomer, 10% dimer and trimer, 30% free fatty acid, and 3% free polylactic
acid. The second composition contains approximately 42% monomer, 16%
dimer and trimer, 30% free fatty acid, and 12% polylactic acid. Three methods
are available to prepare a lactylic ester with a 60% monomer content. The first is
the reaction of lactic acid of known monomer content with a fatty acid chloride
with or without the presence of a weak base to capture the hydrogen chloride
generated (Thompson and Buddemeyer, 1956). This method uses a costly acid
chloride and an extensive purification process. The second method involves the
direct esterification of about a 1:1 molar ratio of sodium lactate and fatty acid,
followed by acidification with mineral acid and isolation of the fatty lactylic ester
(Eng, 1972). A third method of preparing a lactylic ester with a 60% monomer
content is to react lactic acid with acetic anhydride to form the lactyl acetate.
The lactyl acetate is then subjected to an acidolysis reaction with fatty acids in
the presence of a sodium base under high-vacuum conditions. The mixture is
then acidified with mineral acid and the fatty lactylic esters isolated.
The lactylic ester with about 42% monomer and 16% dimer and trimer
20 Food Emulsifiers and Their Applications
acids most probably is prepared by the direct esterification of lactic acid with
fatty acid in about a 1:1 molar ratio (Thompson et al., 1956). A high free poly-
lactic acid content (see Chapter 3 for analytical procedure) indicates that the
product has not been water-washed.
There are no restrictions on the lactic acid content of the lactylic esters of
fatty acids in the FDA regulations (21CFR172). There are no FDA restrictions
on the edible fatty acids that may be used in the preparation of the lactylic es-
ters of fatty acids; however, the stearate esters are most commonly produced.
2.4.1.2 Metallic Salts of Lactylic Esters of Fatty Acids. There are two metal-
lic salts of lactylic esters of fatty acids that are used in the food industry: cal-
cium salt and sodium salt. The calcium salt was commercialized first, as a
dough conditioner for bread, and is known as calcium stearoyl-2-lactylate.
The 2 indicates that 2 moles of lactic acid were used in its preparation. The
sodium salt, which was commercialized later, is known as sodium stearoyl-2-
lactylate, sodium stearoyllactate, or simply "SSL." It is used extensively in
many food applications.
Both esters can be prepared by the same process, the direct esterification of
the partial salt of lactic acid with a fatty acid in about a 2: l molar ratio of lac-
tic acid to fatty acid (Thompson et al., 1956). Both esters have a monomer con-
tent of about 40%, a polymeric content of about 31%, and a polylactic acid
content of about 6%.
Care must be taken to use high-quality starting materials to avoid excessive
darkening during the reaction process. The color of the final product can be re-
duced by the use of hydrogen peroxide as a bleaching agent.
The amounts of lactic acid and metal ion content of both esters are regulated
by the FDA and are about 34% and 4.5%, respectively. Only the use of specified
grades of stearic acid is permitted by the FDA regulations for the products.
OH
0 OH
I
+ CH -C-COOH
3 I
____. r,--o--c I I
-c-cH, + Hp
H CH-OH
(5)
2.4.2.2 Lactylated Fatty Acid Esters of Glycerol and Propylene Glycol. Another
emulsifier, prepared from lactic acid by the reaction of the carboxylic acid
group of the lactic acid with the hydroxyl groups of glycerol and propylene gly-
Synthesis and Composition of Food-Grade Emulsifiers 23
col monoesters, is the lactylated fatty acid esters of glycerol and propylene gly-
col. In this type of emulsifier, propylene glycol is interesterified with an edible
fat or oil to yield a mixture of mono- and diglycerides and propylene glycol mo-
noesters. The reaction mixture is then reacted with lactic acid. As with the
lactylated mono- and diglycerides, a great variety of propylene glycol mo-
noester and monoglyceride mixtures could be used to react with lactic acid to
yield a final composition. In addition, lactic acid can react with any free propy-
lene glycol and free glycerine initially present or formed by a disproportiona-
tion of the starting propylene glycol monoesterlmonoglyceride composition.
These propylene glycol lactates and glycerol lactates are removed by water-
washing or vacuum distillation of the reaction product. Obviously a complex
composition is produced.
It should be noted that only propylene glycol esters prepared by an inter-
esterification process may be used. Propylene glycol mono- and diesters pre-
pared by a direct fatty acid esterification route are not allowed under the
pertinent FDA regulation (21CFR172.850). The FDA regulation further speci-
fies the WICLA content of the final ester composition to be 14 to 18%.
heat
catalyst
OH OH OH
I I
CH 2-cH-cH 2-0-CH 2-CH-CH 2-0H
I Diglycerol
OH OH OH OH
I I I
CH 2-CH-(H 2-0-CH 2-(H-(H 2-0-cH 2-CH-CH 2-0H
I Triglycerol
OH OH OH OH OH
I I I I
CH 2-CH-cH 2-G-CH 2-CH-CH 2-0-CH 2-CH-CH 2-0-CH 2-CH-CH 2-0H
I
Tetragl ycerol
Table 2.2
Theoretical Theoretical
Polyol hydroxyl value molecular weight
Glycerine 1839 92
Diglycerol 1352 166
Triglycerol 1169 240
Tetraglycerol 1072 314
Pentaglycerol 1012 338
Hexaglycerol 971 462
Heptaglycerol 942 536
Octaglycerol 920 610
Nonaglycerol 902 684
Decaglycerol 888 758
Synthesis and Composition of Food-Grade Emulsifiers 25
CH 2-DH CH 2-0H
I I
CH-OH CH-OH
I catalyst I
CH-OH + R-COOH CH-OH +
I I
CH-OH CH-OH
I I
CH-DH CH-OH
0
I I II
CH 2-0H CH 2-0-C-R
CHI
J=l
I
r_j
0
L<JH I
I o
r-<JH I + CH
I
CH_j 0 CH-OH
I I
CH 2-0-C-R
~
CH-OH
I I
Sorbitan Sorbide
monoester monoester
(7)
28 Food Emulsifiers and Their Applications
I
CH-O-CH 2-CH 2-0H CH-Q-CH -CH -OH
I 2 2
I
CH -O-C-R
2
II
0
(8)
Synthesis and Composition of Food-Grade Emulsifiers 29
0 0
II II
CH 2-0-C-CH 2-CH 2-COOH
CH,--c~
I
+ 0 -------+ CH-OH
I
CH -0-C-R CH--c/ CH -0-C-R
2
I 2
II 2
II
0 0 0
Monoglyceride Succinic anhydride Succinylated monoglyceride
(9)
30 Food Emulsifiers and Their Applications
COOH
I
HO-CH
I
HO-CH
I
COOH
(lOa)
32 Food Emulsifiers and Their Applications
CH -0-C-R
2
I
0
COOH
0 /
I
H3 C-c-O-CH
0
I
H3 c-c-o-cz o
""IIc-o CH
2
I
CH-QH
I
DATEM
(lOb)
pH; if the food product is basic, the DATEM ester can be converted to the an-
ionic form. The two acetyl groups are somewhat liable to hydrolysis, which
yields acetic acid. These esters should not be used in aqueous systems that are
expected to have a long storage life.
Two different types of citric acid-based emulsifiers are permitted as direct
food additives: stearyl monoglyceride citrate (21CFR172.755) and monoglyc-
eride citrate (21CFR172.832). The monoglyceride citrate is restricted to use in
antioxidant solutions. It is prepared by the reaction of glycerol monooleate
with citric acid. The final product has an acid number of 70 to 100 and has a
14 to 17% total citric acid content.
Stearyl monoglyceride citrate is prepared by the reaction of citric acid,
monoglycerides, and stearyl alcohol to yield a composition with an acid num-
ber of 40 to 52, a saponification number of 215 to 255, and a total citric acid
content of 15 to 18%. The expected reaction is shown in Equation 11.
As can be seen, the reaction products are quite complex. Both saturated and
unsaturated monoglycerides may be used. This type of ester is used in shorten-
ings containing other emulsifiers and is not used extensively in the food industry.
CH 2-COOH CH-QH
I I
HQ-C-COOH + CH-QH +
I I 0
II
CH 2-COOH CH-0-C-R
(ll)
Note that in direct reaction acetylation, the hydrophilic hydroxyl group is re-
placed by a lipophilic acetyl group; thus, if a fully acetylated ester is prepared,
the product is no longer an emulsifier since it has no hydrophilic groups. In the
interesterification process both fully esterified and partially esterified composi-
tions can also be formed, depending on the amount of glycerine used.
The more fully esterified compositions are useful for their film-forming and
moisture barrier properties.
FDA regulation 21CFR172.828 specifies that the final composition have a
Reichert-Meiss! value of 75 to 200 and an acid value under 6. The Reichert-
Synthesis and Composition of Food-Grade Emulsifiers 35
0 0
I I
+ CH 3-c-o-c-c-CH 3 ------+ +
CH -0-C-R CH -0-C-R
2
I 2
I
0 0
(12)
0 0
I I
CH 2-0H CH 2-0-C-cH 3 CH 2-0-c-R
I I
0
I
+
I ~ catalyst
CH-OH + CH-O-C-cH 3 CH-Q-C-R
0
I I I I
CH 2-0H CH 2-0-C-cH 3
0 0 0
I I I
CH 2-0-C-CH 3 CH 2-0-C-cH 3 CH 2-Q-C-R
I
CH-O-C-CH 3
~ +
I
CH-0-C-R
0
I
+
I
CH-OH
0
I I II I
CH 2-0-C-R CH -0-C-CH
2 I 3
+ + other
compounds
(13)
36 Food Emulsifiers and Their Applications
Meissl number determines the amount of short-chain fatty acids in the final
composition, and this is a measure of the degree of acetylation (see Chapter 3
for methodology). The degree of acetylation in commercially available acety-
lated monoglycerides ranges from 50 to 90%.
HOCH 2
0
0 H H
+
H
HO 0
H OH OH H
Sucrose
0
II
R-C-OCH 3 _______.
Methyl esters
RCOOCH 2
0
H
HO 0
H OH OH H
(14)
produce an emulsion particle size less than one-quarter the wavelength of light,
in contrast to other emulsions in which particle sizes are greater than the wave-
length of light. This particle is so small the emulsion appears transparent.
However, the use of the high amount of the metal salts of the fatty acids and un-
reacted sucrose leads to difficult purification and separation procedures. In gen-
eral, solvents such as ethyl acetate, methyl ethyl ketone, or isobutyl alcohol are
used during the purification process. FDA rule 21CFII 72.859 specifies the
amount of these solvents that can be in the final sucrose ester. A patent has been
issued regarding the purification of the sucrose ester reaction mixture (Mizutani
et al., 1973), but many other purification processes are possible. Overall, the
production of the sucrose monoesters is a more complicated process than the
production of the other food-grade emulsifiers.
References
Babayan, V.K., Lehman, H. (1973). U.S. Patent 3,637,774, January 25.
Barsky, G. (1950). U.S. Patent 2,509,414, May 30.
Brown, K.B. (1943). U.S. Patent, 2,322,820, June 29.
Egan, R.R., Lampost, S.B. (1969). U.S. Patent, 3,433,645, March 18.
Eng, S. (1972). U.S. Patent 3,636,017, January 18.
Federal Register, 53, 40808, December, 1988.
Feuge, R.O., Bailey, A.E. (1946). Oil and Soap, 23, 259.
Food Chemical Codex (1981). 3d ed., pp. 98-99.
Freund, E.H. (1966). U.S. Patent 3,293,272, December 20.
Harris, B.R. (1939a). U.S. Patent 2,177,983, October 31.
- - (1939b). U.S. Patent 2,177,984, October 31.
- - (1941). U.S. Patent 2,258,892, October 14.
Hass, H.B. (1959). U.S. Patent 2,893,990, July 7.
Japanese Patent Publication (1974). No. 15246, April13.
Martin, J.B. (1968). U.S. Patent 3,375,262, March 26.
Mizutani, N., et al. (1973). U.S. Patent 3,798,324, July 24.
Osipow, L.l., Rosenblatt, W. (1969). U.S. Patent 3,480,616, November 25.
- - (1972). U.S. Patent 3,644,333, February 22.
Stockburger, G.J. (1981). U.S. Patent 4,297,290, October 21.
Thompson, J.B., Buddemeyer, B.D. (1956). U.S. Patent 2,744,825, May 8.
Thompson, J.B., eta!. (1956). U.S. Patent 2,733,252, January 31.
Yamagishi, F., eta!. (1974). U.S. Patent 3,792,041, February 12.
THREE
3.1 Introduction
Analytical methods for food emulsifiers are closely associated with or derived
from methods commonly used for fats and oils (Sonntag, 1982; Karleskind,
1996). Test methods are of several types and are carried out for a variety of
reasons. Analysis ensures that the composition of the emulsifier is correct and
that it has not seriously degraded during processing. Often the composition
and distribution of homologs has implications for the utility of the emulsifier in
the individual food product (see, for example, Halkier, 1980). The final level of
testing is often a measurement of performance in the food system itself.
Specifications are negotiated and agreed upon between the producer and the
customer (usually a processed food manufacturer). Analytical tests are carried
out on the process line or control laboratory of the supplier, who then provides
a certificate of analysis to the customer. The customer may then check the
analysis as part of the receiving procedure and accept or reject the shipment.
Disputes may be settled by submitting a sample to an independent laboratory.
Quality assurance testing is also necessary to ensure that foods conform to
39
40 Food Emulsifiers and Their Applications
standards set by government regulatory agencies. The U.S. Food and Drug
Administration (FDA), the European Economic Community (EEC), and the
regulatory agencies of individual countries have limits on the maximum con-
centration of emulsifiers or other food additives allowed in specific products.
This topic has been reviewed by several authors (Kroeller, 1966, 1968;
Gernert, 1968; Amano, 1979). Professional organizations, such as the
Association of Official Analytical Chemists (AOAC), the American Oil
Chemists' Society (AOCS), International Union of Pure & Applied Chemistry
(IUPAC), Leatherhead Food Research Association, and the National Academy
of Sciences (Food Chemicals Codex), have compiled lists of official methods
that are used for fats, oils, and emulsifiers (Cunniff, 1995; Firestone, 1990;
Paquot and Hauffen, 1987; Slack, 1987; Taylor, 1996).
To determine emulsifiers in intact food products, the fats and emulsifiers must
first be extracted. Fats and oils are soluble in nonpolar solvents, such as hexane
or toluene. However, emulsifiers are amphiphilic and more polar and tend to
be less soluble in cold hexane or petroleum ether, particularly when emulsifier
concentration is high compared to total lipid level. Chloroform or chlorofonn/
methanol mixtures (Flor and Prager, 1980) are sufficiently polar but also suffi-
ciently immiscible to allow for extraction from aqueous food systems. In cases
where total lipid content is high compared to emulsifier concentration,
Halverson and Qvist (1974) have reported extracting total lipids with refluxing
hexane followed by extraction with acetonitrile. Solid foods samples (e.g., cake,
coffee whitener) are conveniently extracted using a Soxhlet extraction apparatus.
Liquid samples (e.g., milk, ice cream mix) are usually separated in a separatory
funnel or countercurrent distribution apparatus. Another factor complicating ex-
traction from foods is that lipids may be tightly complexed by starches or pro-
teins or may simply be encapsulated in a biopolymeric matrix. Jodlbauer (1976)
has demonstrated the use of an amylase enzyme to release lipids from a pasta
product so that they are more easily extracted.
3.3.1 a- Monoglyceride t
Monoglycerides may exist as alpha, the most predominant form, which has the
fatty acid esterified in the 1 or 3 positions of the glycerol head group, or beta,
where the substitution is at the 2 position. The alpha isomer has adjacent hy-
droxyl groups in the 1, 2 or 2, 3 positions. The isomer is determined by titration
with periodic acid resulting in cleavage, as shown in the following equation:
The analysis is carried out by dissolving the sample (extracted lipid or pure
OH
0
II
CH -c-c
~ 2 H I
0 -~-~C17 H35 + HI04 + HI03 + Hp
0
(15)
(A- B)(N)(56.1)
w
where A mL standard alkali used in titration
B = mL standard alkali used to titrate blank
N = normality of standard alkali
W = g of sample
Free fatty acid percentage can be determined by dividing the acid value by
a factor characteristic of the fatty acid present. Values for this factor are C12
(lauric)= 2.81, C16 (palmitic)= 2.19, and C18 (stearic or oleic)= 1.99.
(S- B)(N)(l2.69)
w
where s= mL used to titrate sample
B = mL used to titrate blank
N = normality of thiosulfate solution
w= weight of sample
In this test, when reporting the value, it is important to also identify the
method by which the value was determined.
Iodine value may also be calculated from the fatty acid composition (FAC) of
fats, oils, and fatty acids through gas-liquid chromatography (GLC)(Firestone,
1990h). The same method may be used for food emulsifiers, but new conversion
factors would probably need to be developed to translate GLC peaks to iodine
values. Near infrared reflectance (NIR) has also been developed as a rapid
method to determine unsaturation by measurement of the carbon-carbon double
bond absorbance peak. However, a great deal of work is necessary up front to as-
semble a calibration curve to relate the measured absorbances to traditionally
measured values.
(V)(T) (1000)
m
Since the practicing formulator needs to minimize the peroxide content of in-
gredients, the lower the peroxide value, the less is the risk of developing rancidity.
Special surveillance should be maintained on materials that have higher iodine
values (higher unsaturation). Of course, other materials that catalyze oxidation,
such as iron, copper, and lipoxygenase enzymes, must also be considered.
Recently, peroxides have been determined utilizing high-performance liq-
uid chromatography (HPLC) (Yang, 1992; Yang et al., 1991). A sample is sep-
arated on a column and fractions then pass through a post-column reaction
chamber where they are mixed with a luminescence reagent. Light emitted
from the chemiluminescence reactions is then measured by a special photo-
chemical detector. The method is fairly rapid and requires only small amounts
of sample. An additional advantage is that different peroxide species may be
measured separately, which may indicate where oxidation is occurring. The
major disadvantage is the capital cost of the equipment and the high cost of the
luminescence reagent.
This value is sensitive to both the degree of esterification and the chain
length of the substituent fatty acid. Higher degrees of esterification and shorter
chain lengths are reflected by higher saponification values. Conversely, longer
chain lengths and lower degrees of substitution are reflected in lower saponifi-
cation values.
_(5_6._1)_(7)_(V._
0 -_V_) + AV
up the determination. Hydroxyl values are used mainly to specify and control
polyglycerols and polyglycerol esters.
t Taylor (l996a).
50 Food Emulsifiers and Their Applications
the short-chain acid in alkali followed by neutralization with dilute sulfuric acid,
(ii) distillation of the liberated acetic (or other short-chain) acid, and (iii) determi-
nation of the condensed acid by titration with standard base to the phenolphthalein
endpoint. The method is therefore equipment intensive (the condensing unit must
be replaced or cleaned between samples) and time-consuming.
3.3.9 Moisture
Moisture is generally undesirable in food emulsifiers because it is a diluent and
it represents a potential problem for microbial growth. Some exceptions are the
polysorbates that require a small amount of water to provide clarity and prevent
phase separation, and emulsifier hydrates where gel phases are preformed for
specific applications. Several methods for moisture have been developed through
the history of oil processing. The distillation method uses a toluene solvent to as-
sist distillation of water from a large sample through a condenser and into a grad-
uated separation tube (Firestone, l990i). The water is quantitated by volume.
The method is equipment intensive because of the need to clean and replace the
distillation apparatus between samples. The distillation method is most
amenable where the water concentration is about 0.5% or higher.
Lower levels of moisture are readily analyzed by reaction with Karl Fischer
reagent. The procedure is amenable to lecithin as well as other emulsifiers that
can be solubilized for the analysis (Firestone, l990p). Instruments are cur-
rently available that allow repetitive determinations without the necessity of
disassembly and cleaning between samples.
3.3.10 Color
Color of emulsifier is a reflection of the quality of starting lipid in its manufac-
ture and/or manufacturing conditions. Thermal abuse of fats and emulsifiers
are well known to produce dark colors. In many cases, oxidized tocopherols
are thought to be the chromatic impurities. Dark-colored emulsifiers may de-
tract from the appearance of finished food products where they are used in
high concentrations (e.g., cake icings). However, color evidence that an emul-
sifier has been abused also suggests that pro-oxidants may be present that may
adversely affect the flavor of the product. Emulsifiers would therefore be
processed to produce the lightest color possible.
t Firestone (l990m).
Analysis of Food Emulsifiers 51
the lecithin obtained in this way is therefore a major concern. Its chemical
structure is that of a phospholipid where phosphoric acid is esterified to a glyc-
erol molecule and an organic base. Total phosphorus in the sample may be de-
termined by a laborious process in which the sample is saponified, phosphorus
is separated by precipitation with molybdate solution, the precipitate is dis-
solved in base, and excess base is titrated with standard nitric acid (Firestone,
1990r). Tamanaka and Kudo (1991) have reported a rapid method where phos-
phorus precipitated by molybdate is determined spectrophotometrically. A
much simpler, albeit less precise, procedure is to determine lecithin content as
acetone-insoluble material based on the principle that neutral lipids are solu-
ble in acetone while phospholipids are not (Firestone, 1990g). This principle
was used by Goldstein (1984) to develop a method for determination of lecithin
in oils. The oil sample was dispersed in acetone and lecithin was quantitated
by measurement of the turbidity.
Individual phosphatides, such as phosphatidylcholine and phos-
phatidylethanolamine may also be determined by HPLC (see Section 3.5,
Instrumental Methods). Combinational measurements have been reported for
separation and phospholipid class identification in commercial soy lecithin
(Lendrath et al., 1991) and in egg yolk (Holopainen, 1972). Individual phos-
pholipids of soybean and rapeseed were separated by micellar electrokinetic
capillary chromatography (lngvardsen and Michaelsen, 1994).
Determining the presence and composition of phospholipids in foods is an
exercise in separation followed by measurement. Wewala and Baldwin (1982)
describe a procedure for the analysis of lecithin in instant milk powders.
Ugrinovits (1983) described determination oflecithin in cocoa powders.
3.4.2 Viscosity/Consistency
Viscosity is a property of an emulsifier that is important to processing and transfer
of material, such as pumping through product feed lines. Measurements are not
generally made routinely. A viscosity/temperature profile is usually constructed,
and acceptable ranges are defined for processing operations. Some classes of
emulsifiers, such as polyglycerol esters, are very thick and have to be heated to be
pumped through heat-traced lines. Sucrose stearate poses a problem since it de-
composes when heated too high and is therefore preferred as a powder form.
Viscosity of lecithins had been measured by a procedure known as the "bub-
ble time" method (Firestone, l990v) that was also used widely in the paint indus-
try. A clear liquid sample is placed in an ASTM tube in a constant-temperature
bath. The tube is inverted to produce a bubble at the bottom of a column ofliquid.
The time for the bubble to reach the top is measured and recorded. The value in
poise is obtained by comparison with standard samples. Viscosity may also be de-
termined directly using a Brookfield viscometer (Firestone, 1990u). In the latter
case, it is not as critical that the samples be clear.
Rheological properties are most often determined when emulsifiers are incor-
porated into foods and ingredients such as shortenings. Consistency (Firestone,
l990e) may be measured using a penetrometer, such as an lnstron or Stevens
texture analyzer. The method measures the distance a probe penetrates into a
sample at a given force, temperature, and time. This property is critical to perfor-
mance when the product is mechanically incorporated into a food product. An
example is dough-sheeting to produce Danish pastries or pizza crust.
t Christie (1989).
56 Food Emulsifiers and Their Applications
PGMP
PGMS
GMO&GMS
GMS
12 15 18 21 24
Figure 3.1 GLC separation of monoglycerides and propylene glycol ester emulsifiers.
(a) Commercial emulsifier; (b) in shortening. (Hasenhuettl et al., 1990.)
Analysis of Food Emulsifiers 57
1. Cholesterol 1. Cholesterol
2. PalmHic acid 2. Phosphalidylethanolamine
3. Phosphatidylethanolamine 3. Phosphalidylcholine
4. Phosphatidylserine 3 4. Sphingomyelin
5. Phosphatidylcholine
6. Sphingomyelin
12 16 2DIIin. 12 16 2D Min.
also been reviewed by Christie (1996). Garti and Aserin (1981) reported the
separation of polyglycerol mono- and polyesters by HPLC on a LiChrosorb col-
umn. Sorbitan esters of fatty acids could also be separated on the same station-
ary phase (Garti and Aserin, 1983). Murakami and coworkers (1989)
described the separation of sucrose esters by preparing their 3,5-dinitroben-
zoyl derivatives. This technique was used to determine sucrose esters in a
wide range of food products.
The development of facile extraction and rapid sample prepurification
methods as well as autosampling technology will undoubtedly encourage more
widespread use of HPLC.
tain signals. These signals are dependent on the environment of the nuclei and
are therefore useful for determining functional groups and chemical structure.
Relaxation time of the signal is useful to determine the physical state of a mol-
ecule. Wide-line NMR is widely used to measure the amount of solid fat or
solid fat content (SFC) in a sample. However, this technique is rarely used for
food emulsifiers and is not included here. Chemical shifts have been utilized
to determine the mesomorphic form of emulsifiers in aqueous systems, which
is likewise beyond the scope of this chapter. The reader is referred to
Lindblom (1996) for a thorough treatment of this subject. Mesomorphic phase
behavior in food emulsions will be discussed in Chapter 6.
Proton (1 H) NMR is the oldest technique. However, because of the large num-
ber of similar protons in alkyl chains of emulsifiers, this technique has not shown
widespread utility. Press and coworkers (1981) observed the protons on choline at
3.3 ppm to determine phosphatidylcholine content in lecithin. Sheeley et al.
(1986), in the same laboratory, proposed that the vinylic protons at 5.3 ppm could
serve as an alternative measurement of the iodine value, discussed earlier.
Chemical shifts of carbon (13 C) are useful for determination of the func-
tional environment of the carbon atom being observed. Since lipids have sig-
nificantly fewer carbons than hydrogen atoms, the problem of spectra
interpretation is more tractable. Gunstone (1993) has recently reviewed the
field of 13 C NMR of lipids. Chemical shifts of isopropylidene derivatives of
monoglycerides have been reported by Dawe and Wright (1988). Sacchi and
coworkers (1990) utilized the shifts of glyceryl and carbonyl carbons to mea-
sure the levels of monoglycerides, diglycerides, and free fatty acids in olive oil.
Chemical shifts for glyceryl carbon atoms are listed for monoglycerides, propy-
lene glycol esters, acetylated monoglycerides, phosphatidylcholine, and phos-
phatidylethanolamine in Table 3.1.
Chemical shifts of phosphorus atoms (3 1P) are of great utility in determining
the structure and concentration in phospholipids. Since there is only one phos-
phorus atom per lipid molecule, assignment is more straightforward than for
1H or 13C NMR. However, this is somewhat offset by the numerous types of
phospholipid molecules found in natural samples, such as egg. Glonek and
Merchant (1996) have recently reviewed this method.
identified from the spectra. An electrospray technique has also been found to be
useful for analysis of sucrose fatty acid esters (Schuyl and van Platerink, 1994).
This technique showed a series of molecular ions that corresponded to the de-
gree of substitution of fatty acids onto the sucrose molecule.
References
Amano, H., (1979). Tekisuto-Zeminaru, 26th Conference, Tokyo: Nippon Yukagaku
Kyokai, 59-74.
Biacs, 0., et al. (1978). Acta Aliment. Acad. Sci. Hung., 7(3):181-93, CA 89:213672.
Birkel, T., et al. (1979). ]. Assoc. Off. Anal. Chem., 62(4):931--6.
Blum, J., Koehler, W. (1970). Lipids, 5(7):601--6.
Bruemmer, J.M. (1971). Brot Gebaeck, 25(11):217-20.
Brueschweiler, H. (1977). Mitt. Geb. Lebensmittelunters. Hyg., 68(1):46-63.
- - , Dieffenbacher, A. (1991). Pure Appl. Chem., 63(8):1153--62.
- - , Hautefenne, A. (1990). Pure Appl. Chem., 62(4):781-93.
Bruns, A. (1988). Fett Wiss. Technol., 90(8):289-91.
Christie, W.W. (1996). "Separation of phospholipid classes by high performance liquid
chromatography," in Advances in Lipid Methodology-Three (ed. W.W. Christie),
The Oily Press, Ayr, Scotland, pp. 77-108.
- - (1992). "Detectors for high performance liquid chromatography of lipids with
special reference to evaporative light scattering detection," in Advances in Lipid
Methodology-One (ed. W.W. Christie), The Oily Press, Ayr, Scotland, pp. 239-271.
- - (1989). Gas Chromatography and Lipids: A Practical Guide, The Oily Press,
Ayr, Scotland.
Cunniff, P. (ed.) (1995.) Official Methods of Analysis of AOAC International, 16th ed.,
AOAC International, Arlington, VA.
Ibid. (1995a). 41.1.63 Method 969.34.
Ibid. (1995b). 41.1.60 Method 942.19.
Ibid. (l995c). 41.1.20 Method 940.28.
Ibid. (1995d). 41.1.18 Method 920.160.
Ibid. (1995e). 41.1.15 Method 993.20.
Ibid. (19951). 4l.l.l2 Method 965.32.
Ibid. (1995g). 4l.l.l6 Method 965.33.
Daniels, D.H., et al. (1985). ]. Agric. Food Chem., 33(3):368-72.
- - (1982). ]. Assoc. Off. Anal. Chem., 65(1):162-5.
Dawe, R.G., Wright, J.L.C. (1988). Lipids, 23(4):355-8.
Dick, R., Miserez, A. (1976). Mit. Geb. Lebensmittelunters Hyg., 67(4):472-87.
Dieffenbacher, A., et al. (1988). Rev. Fr. Corps. Gras, 35(12):495-9
- - (1989). Rev. Fr. Corps. Gras, 36(2):64.
Duden, R., Fricker, A. (1977). Fette Seifen Anstrichm., 79(12):489-91.
El-Sebaiy, L.A., eta!. (1980). Food Chem., 5(3):217-28.
Erdahl, W.L., et al. (1973). ]. Amer. Oil Chemists Soc., 50(12):513-15.
64 Food Emulsifiers and Their Applications
Carbohydrate/Emulsifier
Interactions
Lynn B. Deffenbaugh
4.1 Introduction
Many different types of emulsifiers are used in food products. The inclusion rate is
usually low, < 3%. Composition and characteristics of emulsifiers used in foods
are discussed in other chapters. Emulsifiers often function indirectly in foods by
modifying the properties of major components such as carbohydrates, protein, or
fat. This chapter discusses the impact of emulsifiers on carbohydrates.
Carbohydrates are a large group of diverse food components. Carbohydrates
can be classified into three categories based on degree of polymerization: mono-
and disaccharides, oligosaccharides, and polysaccharides. Polysaccharides
are further divided into starch and nonstarch fractions. The nonstarch polysac-
charides are plant cell wall materials that contribute dietary fiber to foods.
Mono-, di-, and oligosaccharides and nonstarch polysaccharides (fiber) inter-
act minimally with emulsifiers from the perspective of food-product character-
istics. Starches, however, form complexes with emulsifiers that significantly
alter the characteristics that the starches provide in foods. Emulsifiers that
complex with starch are notably different in function and usage from protein-
complexing emulsifiers discussed in the next chapter.
This chapter discusses how complexes form between starches and emulsi-
67
68 Food Emulsifiers and Their Applications
The long axis of complexing agents is internal and parallel to the long axis
of the amylose molecule (Mikus et al., 1946; Rundle and French, 1943a, b).
Complexing agents interchange reversibly, competing for the same space in
the amylose helix (Mikus et al., 1946; Schoch and Williams, 1944). The unit
cell packing size and distance between starch helices are not altered by the
type of complexing agent (Raphaelides and Karkalas, 1988).
Lipid chains usually occupy the helix as paired dimers linked by hydrogen
bonds at the carboxyl groups in fatty acids or glycerol in monoglycerides
(Raphaelides and Karkalas, 1988). The hydrocarbon chains in complexes are so
highly ordered that they have been reported to be crystalline (Carlson et al., 1979).
Amylose helices induced by complexation aggregate into partially crys-
talline structures that exhibit a V-type x-ray diffraction pattern (Szczodrak and
Pomeranz, 1992) and are insoluble. These insoluble complexes consist of
lamellar crystals with the helix perpendicular to the lamella (Raphaelides and
Karkalas, 1988). Amylose and amylopectin complexes can be differentiated by
their physical properties. Amylopectin emulsifier complexes are more soluble
than amylose complexes. The insolubility of the fatty acid/amylose complex
has long been used as a means to selectively precipitate amylose, but not amy-
lopectin, from solution (Schoch and Williams, 1944). Complexes cannot al-
ways be assumed to be insoluble, however, since the degree of insolubility
varies with the surfactant (Kim and Robinson, 1979). The complex between
starch molecules and iodine is used to differentiate and quantitate the amylose
fraction, which forms a blue complex, and the amylopectin fraction, which
forms a red-purple complex.
4.3.3 Starch-Pasting
Starches and starch-containing ingredients provide many of the physical and
organoleptic properties of foods. Dramatic changes occur in starches and
starch-containing products when processed. When starches are cooked in the
presence of water, starch granules absorb water and swell to many times their
original size. Amylose, the linear fraction of starch, leaches from the granules.
A hot starch paste is a mixture of swollen granules, granule fragments, and col-
loidally and molecularly dispersed starch molecules (Olkku and Rha, 1978).
The viscosity increases greatly as this hot paste matrix forms. The fragile,
swollen granules will eventually disintegrate, especially when shear is ap-
Table 4.1 Effect of emulsifiers and complexing agents on properties of nonwaxy starch
CSL =calcium stearoyl-lactyl-2-lactylate; CTAB =cetyltrimethylammonium bromide; DATEM =diacetyl tartaric acid esters of monoglylcerides; EMG =ethoxylated
monoglycerides; GMP =glycerol monopalmitate; GMS =glycerol monostearate; MG =monoglycerides; POEMS =polyoxyethylene stearate; SDS =sodium dodecyl
sulfate; SLS =sodium Iaury! sulfate; SSL =sodium stearoyl-2-lactylate.
Table 4.2 Effect of emulsifiers and complexing agents on properties of waxy starch
.......
Ul
~ Table 4.2 continued
CSL = calcium stearoyl-lactyl-2-lactylate; CTAB = cetyltrimethylammonium bromide; DATEM = diacetyl tartaric acid esters of monoglycerides; GMS = glycerol
monostearate; MG = monoglycerides; POEMS= polyoxyethylene stearate; SDS =sodium dodecyl sulfate; SLS =sodium Iaury! sulfate; SSL =sodium stearoyl-2-lacty-
late.
Carbohydrate/Emulsifier Interactions 77
.--..
1.0
...::::
()
L..
.....0en 0.8
0> •--•Hylon VII
E o--oWheat
...._,
IJ") 0.6 A - - A Maize
c--o Potato
E o--oTapioca
c:
0
0.4 v--vWaxy Maize
0
.....
tO
0 0.2
en
..0
<(
T--------v-----~v------------------------------v
0.0
0 2 3 4 5
% SE (starch wt. basis)
plied. This relative decrease in viscosity after time and application of shear
defines a peak viscosity.
The process whereby the ordered structure of the starch granule is lost is
called "gelatinization." The transition is a first-order water-mediated melting
of crystalline regions in the starch granule (Donovan, 1979; Zobel, 1984).
Maximum solubilization and swelling occur when excess (> 5 times) water is
present, typical of sauces, gravies, and puddings. In lower moisture systems,
such as baked or extruded products, granule swelling can be limited by mois-
ture content. Extremely high viscosities result because diluent is limited.
Uses and functions of starches are multiplied by modifying properties with
emulsifiers. For example, time of addition and/or time for diffusion of fatty ad-
juncts into starch granules are processing parameters than can be varied to
produce substantially different properties in cooked starch or cereal grain
products (Lund, 1984). If added before starch gelatinization, monoglycerides
penetrate the granule, form molecular complexes, and reduce starch swelling
power. Addition of monoglycerides after gelatinization enhances granule sta-
bility (van Lonkhuysen and Blankestijn, 1974). The function of starch-com-
plexing emulsifiers is primarily due to formation of starch/emulsifier
complexes. The magnitude of the effect of starch/emulsifier complexes on
starch cooking properties varies with starch type, emulsifier type, and process-
ing conditions.
78 Food Emulsifiers and Their Applications
Emulsifier and starch molecules must have access to each other to interact.
Starch-complexing emulsifiers must be soluble enough or in a phase that
yields monomers (see Section 4.4.4). Starch molecules have limited availabil-
ity until gelatinization begins. Emulsifiers adhere to the granule surface before
gelatinization, and they begin to form insoluble starch/emulsifier complexes as
soon as the granule begins to swell and amylose begins to solubilize. These in-
soluble complexes near the surface stabilize the granule. The rate of further
swelling and amylose leaching are slowed. Gelatinization temperature is in-
creased because more energy is required to cook or swell the stabilized gran-
ule. Some emulsifiers, such as polysorbate 60, may cover the starch surface
with a film, increase hydrophobicity, and inhibit water transfer into the granule
(Kim and Walker, 1992c).
The effect of emulsifiers on starch gelatinization is detected in many ways
(Table 4.1). When starch pastes were made with SSL or GMS, changes in vis-
coelastic properties of the paste were coincident with reduced granule swelling
(Eliasson, 1986b). The granules were less deformable (stiffer), as indicated by
an increase in the temperatures where peaks in storage (G') and loss (G") mod-
ule were reached. Pasting temperature, hot viscosity, and temperature of peak
viscosity of most normal starches are increased in the presence of emulsifiers
that have a fatty adjunct that can form inclusion complexes with starch.
According to Mitchell and Zillman (1951), the increase in starch paste viscos-
ity in the presence of complexing agents is a result of the increased ability of
granules to absorb and hold water without rupturing.
Gelatinization, the melting of crystalline regions in the granule, causes loss
of birefringence and disappearance of a starch x-ray diffraction pattern
(Eliasson, 1986a). When emulsifiers delay gelatinization, the loss of birefrin-
gence is delayed (see Table 4.1).
Some emulsifiers with a fatty acid chain adjunct do not form complexes
with starch. Anionic surfactants, such as sodium lauryl sulfate (SLS) and
sodium dodecyl sulfate (SDS), destabilize the granule because of its highly
negative charge (Eliasson, 1986b; Moorthy, 1985). The destabilization leads to
rapid, complete swelling of the granules, and a rapid increase in viscosity
early in the pasting (cooking) cycle. Loss of granule integrity will subsequently
lead to a dramatic loss in viscosity after peak. This is undesirable if the starch
is being utilized for its thickening, texturizing abilities. In the selection of an
emulsifier for product development applications, charge as well as chemical
composition need to be considered when effect on starch is important.
Carbohydrate/Emulsifier Interactions 79
4.3.5 Retrogradation
The primary reasons for using emulsifiers are modification of finished product
characteristics. Retrogradation in starch-containing foods impairs texture and
taste, and emulsifiers inhibit this retrogradation.
Retrogradation is the formation of ordered, partially crystalline sites within
a cooled starch paste or gel. The recrystallization of starch is a long-term
process that occurs during the hours and weeks after pasting and gelation.
Amylose retrogrades so fast that it is often complete before the food is con-
sumed. Retrogradation of amylopectin occurs more slowly and affects the tex-
ture and taste offoods throughout shelf life (Miles et al., 1985).
Emulsifiers decrease the rate of retrogradation of starch gels mainly through
emulsifier/starch-complex formation (Miura et al., 1992). The helix structure of
starch in a complex prevents side-by-side stacking of starch molecules and,
80 Food Emulsifiers and Their Applications
200
0 ~~--~-+--~~-+_,
0 4 6 B I 0 IZ I< 16 18 20 16 IB 20
Time (n1in)
I5>0°CI H.ot to g~0c I c~ lo S0°C
120
: h
100
- w,.,.. .
I / - · - · -• - • - · - •
100
! i
g eo ~: ~~ ;~\:... ~~/-·-·--~
1:-
8 60 ·g GO ;.\\ \.. //1
5
Q
.'-
40
5•
...0 40
j \~:~. ]!
0
1 i:
~ 20 : 20 I i:
,_{.~-!:,'
0~~-+--~+--l~r-- ~-- ·
0 8 10 IZ 14 16 18 20 8 10 12 14 IG 18 20
Time (min} Timl!!(mln}
I50°CI Heat to 95°C I Cool to ~C Heat to 95°C I Cool lo .50°C
Figure 4.2 Rapid Visco Analyzer viscosity profiles of maize, potato, tapioca, and
wheat starches with 0, 1, 2, or 5% (starch wt basis) of a sucrose ester emulsifier. (From
Deffenbaugh, J990a .)
4.3.6 Enzymolysis
Complex formation interferes with the ability of enzymes to digest starch molecules
(see Table 4.1). Kinetics of glucoamylase, which cleaves successive glucose units
from the nonreducing end of starch chains, were not directly affected by a sucrose
ester emulsifier (Deffenbaugh, 1990a). Reduced digestibility of normal starches in
the presence of an emulsifier were due to decreased availability of the starch sub-
strate. Precipitation of the complex and reduced steric compatibility of helical
starch with the active site of the enzyme reduced the starch substrate availability.
Glucoamylase digestion of waxy maize starch was not inhibited by a su-
crose ester (Deffenbaugh, 1990a). If a complex formed in the outer branches, it
was not stable enough or strong enough to inhibit glucoamylase.
The inhibition of starch enzymolysis by emulsifiers is analytical evidence of
complex formation. It should be noted however, that emulsifier/starch-complex
formation does not have a significant effect on overall extent of digestibility of
starch in vivo as shown in rats (Holm et al., 1933).
has been reported with 18-carbon saturated fatty acids (Hahn and Hood, 1987)
and 14- to 16-carbon fatty acids on monoglycerides (Hoover and Hadziyev,
1981; Krog, 1971; Lagendijk and Pennings, 1970). Geometrical structure as
well as chain length and molecular weight will influence the ability of the lig-
and to penetrate the starch helix (Miura et al., 1992). If chain length as well as
helix conformation are known, the fatty acid to amylose ratio required to
achieve saturation of the amylose helix is predictable from stoichiometric rela-
tionships (Karkalas and Raphaelides, 1986).
Solubility of the adjunct affects the equilibrium between complex formation
and concentration of the adjunct in solution. The lower solubility of fatty acids
versus monoglycerides in aqueous systems drives fatty acids into the lipophilic
core of the amylose helix more than monoglycerides.
Increased unsaturation in free fatty acids or fatty acid chains of monoglyc-
erides reduces the ability of the adjunct to bind with starch (Hahn and Hood,
1987; Krog, 1971; Lagendijk and Pennings, 1970). The 30° bend at cis bonds
inhibits rotation and causes steric hindrance. In addition, lower solubility of sat-
urated fatty acids will favor complex formation with the hydrophobic helix core.
Steric hindrance from bulky groups on emulsifiers and surfactants interferes
with the ability of the adjuncts to form complexes with starch (Gray and Schoch,
1962). Monoglyceride binding is less than binding of fatty acids of the same
chain length because steric hindrance of the glycerol group interferes with com-
plex formation (Hahn and Hood, 1987). Propylene glycol esters of fatty acids,
with a particularly bulky polar group, have lower complexing ability than emul-
sifiers with smaller polar groups (Krog, 1971). In addition to steric hindrance,
hydrophilic polar groups are not compatible with the hydrophobic helix core.
The higher the proportion of the polar group to the overall molecule, the less
compatible the molecule is with the hydrophobic interior of the helix.
talline complex, increased when held at 60°C for 1 hour. These authors con-
cluded that the surfactants do enter the granule and complex with amylose be-
fore gelatinization.
I, 2, 3, 4 These numbers indicate significant difference (P < 0.05) within starch type.
dimers at lower pH. pH does not affect binding of monoglycerides because the
carbonyl group of the fatty acid is involved in the ester linkage to glycerol.
Amylose-complexing ability of compounds containing hydrocarbon chains
is significantly affected by the phase behavior of the lipid material (Larsson,
1980). The most effective amylose-complexing emulsifiers have a high degree
of molecular freedom in an aqueous phase and exhibit lyotrophic mesomor-
phism. Micelles or liposomes of emulsifiers are the most effective polymorphs
for providing lipid monomers, and as a result, are more effective complex for-
mers than lipids in other polymorphic states (Riisom et al., 1984; Eliasson,
1986a). Lysolecithin, a native lipid in cereal starches, complexes readily with
Carbohydrate/Emulsifier Interactions 85
that the coils of the helix are in contact with each other (French and Murphy,
1977). Jane and Robyt (1984) studied the lamella-like organization of V-type
complexes and concluded that starch chains were folded such that the chain
axes were perpendicular to the surface of the lamella.
and water (Kugimiya et al., 1980). The net enthalpy of starch gelatinization en-
dotherm is reduced in the presence of external lipids due to simultaneous,
exothermic crystallization of starch/lipid complexes (Biliaderis et al., 1986;
Deffenbaugh, 1990a; Eliasson, 1983; Eliasson, 1986a; Eliasson and Ljunder,
1988; Eliasson et al., 1988; Evans, 1986; Kugimiya et al., 1980). Data is given
in Table 4.4 showing the effect of a sucrose ester emulsifier on gelatinization
temperature and enthalpy of various starches (Deffenbaugh, 1990a). Microscopy
and viscosity profile measurements, discussed previously, also detect a delay in
gelatinization of normal starches in the presence of emulsifiers. Depending on
moisture and emulsifier levels, DSC may not be sensitive enough to detect the
delay in starch gelatinization caused by emulsifiers (Deffenbaugh, 1990a; Kim
and Walker, 1992b). See Table 4.4.
To eq
Starch 0% SE 1% SE 2% SE 5% SE
4.6 Summary
It is ironic that the interaction between emulsifiers and starches is "simply"
based on one phenomenon-the formation of inclusion complexes between
starch helices and fatty adjuncts. The amount of research devoted to under-
standing the interaction between starches and emulsifiers is daunting. Many
methods and measurements are used in these studies because each provides a
different perspective to understanding the extent and nature of the interaction.
The physical and sensory characteristics of many foods are largely defined by
starches. New and improved foods are developed because emulsifiers can be
used to modify the characteristics of starches in foods. Reduction of stickiness in
reconstituted potatoes is an example. The food industry is also dependent on
emulsifiers to improve shelf life of foods. The economic advantage of increasing
shelf life of bread with anti staling emulsifiers outweighs the cost of the emulsifier
by manyfold. Applications for starch-complexing emulsifiers will continue to ex-
pand as the properties of the interaction are better understood.
Acknowledgment
The author acknowledges with great appreciation the assistance of Mrs. Teresa
Booher in preparation of the manuscript.
References
Banks, W., Greenwood, C.T. (1975). Starch and Its Components, Edinburgh University
Press, Edinburgh, Scotland.
Batres, L.V., White, P.J. (1986). Interaction of amylopectin with monoglycerides in
model systems,}. Am. Oil Chem. Soc., 63, 1537-40.
Biliaderis, C.G., Galloway, G. (1989). Crystallization behavior of amylose-V complexes:
structure-property relationships, Carbohydrate Res., 189, 31-48.
Carbohydrate/Emulsifier Interactions 91
Protein/Emulsifier Interactions
Martin Bos, Tommy Nylander, Thomas Arnebrant,
and David C. Clark
5.1 Introduction
Many food emulsions are more complex than the traditional definition of an
emulsion: a colloidal dispersion of liquid droplets in another liquid phase.
This is mainly because the dispersed phase is often partially solidified or the
continuous phase may contain crystalline material, as in ice cream. However,
one characteristic that all emulsions have in common is that they are (thermo-
dynamically) unstable. The four main mechanisms that can be identified in the
process of breaking down an emulsion are creaming, flocculation, coalescence,
and Ostwald ripening. There are two ways in which the process of breakdown
of an emulsion can be influenced. First, use of mechanical devices to control
the size of the dispersion droplets and second, the addition of stabilizing
chemical additives like low molecular weight emulsifiers and polymers to keep
it dispersed. The main purpose of the latter is to prevent the emulsion droplets
from fusing together (coalescence), often achieved by repulsive droplet/droplet
interactions. These interparticle interactions are determined mainly by the
droplet surface, which is coated with emulsifiers, often biologically surface-ac-
tive components like proteins, mono- and diglycerides, fatty acids, or phospho-
lipids. The forces most commonly observed are electrostatic double layer, van
95
96 Food Emulsifiers and Their Applications
der Waals, hydration, hydrophobic, and steric forces. They are responsible for
many emulsion properties including their stability.
The complex mechanisms involved in formation, stabilization, and destabi-
lization of emulsions make fundamental studies on applied systems difficult.
One approach has therefore been to clarify the basic physical and chemical
properties of emulsions by the study of simpler model systems. The adsorption
behavior of single-emulsion components like proteins, fatty acids, surfactants,
or phospholipids at liquid/air or liquid/liquid interfaces have given informa-
tion about surface activity, adsorbed amounts, kinetics, conformation, and sur-
face rheology. The development of experimental techniques has made it
possible to extend these studies to multicomponent systems. This has provided
further information concerning competitive adsorption, displacement, and
complex formation, which can be related to emulsion and foam stability.
For further information concerning the physicochemical factors affecting
the emulsion structure as well as characterization of food emulsion stability,
the reader is referred to the reviews of Dickinson (1987a; 1988), Dickinson
and Stainsby (1982), and Tadros and Vincent (1983), and for the principles of
emulsion formation to the review of Walstra (1983) along with the other chap-
ters in this book. In this chapter we will focus on the molecular interactions
between proteins and other surface-active components present at the interface
of the emulsion droplets that prevent the droplets from fusing together.
Understanding the interaction between these emulsifier components is the key
to increase the emulsion stability as well as to be able to tailor the structure of
these systems. Various surface-active components like lipids, low molecular
weight (LMW) surfactants, and even phospholipids will be regarded as emulsi-
fiers. We will first discuss the stability of the protein in liquid solutions, which
is an important factor for their behavior in emulsion systems. Although the be-
havior at liquid/liquid and liquid/air interfaces can be best compared with the
situation in an emulsion or foam, we will also discuss some relevant studies
concerning the solid/liquid interface as well as the effect of emulsifiers on the
solution behavior of proteins. A number of techniques can be used to study
protein/emulsifier interactions, including surface film balance, ellipsometry,
Brewster angle microscopy (BAM), circular dichroism (CD), differential scan-
ning calorimetry (DSC), surface rheology, fluorescence spectroscopy, and neu-
tron reflectivity. It is beyond the scope of this chapter to discuss these
techniques in detail, but when necessary a brief explanation will be given.
The link between the molecular interactions between emulsifier compo-
Protein/Emulsifer Interactions 97
nents and the properties of food emulsions will be discussed in the last section
of this chapter.
a mean packing density of 0.74, which is comparable with the values found for
crystals and organic molecules (Richards, 1977). In this densely packed envi-
ronment, short-range forces like van der Waals interactions and hydrogen
bonding play an important role (Privalov, 1978). In addition, the hydrophobic
interactions contribute significantly in stabilizing the structure, as first pointed
out by Kauzmann (1959). Accumulation of hydrophobic residues in the core of
the molecule leads to a strong folding force. In contrast, the polar groups prefer
the outside of the protein molecule due to their solvent affinity.
Due to the delicate balance of forces mentioned above, the protein is margin-
ally stable (Pace, 1981; Privalov, 1988; Dill, 1990). Changing one amino acid
residue might stabilize or destabilize the protein (Matsumura et al., 1988).
Binding of a lipid, surfactant, or another protein molecule can also have the
same effect. For instance, it has been shown that at low surfactant-to-protein ra-
tios, the binding can stabilize the protein against thermally induced unfolding,
as observed for the interaction between fatty acids or sodium dodecylsulfate
(SDS) and bovine serum albumin (Gumpen et al., 1979), as well as SDS and~
lactoglobulin (Hegg, 1980). On the other hand, increasing the surfactant-to-pro-
tein ratio above lO moles of SDS per mole serum albumin or 1 mole of SDS per
mole of J3-lactoglobulin monomer causes unfolding of the protein. Creamer
(1995) observed similar stabilization of ~-lactoglobulin against urea induced un-
folding at 1:1 molar ratios between the protein and SDS as well as palmitate. In
many emulsions involving proteins and emulsifiers, the interaction could take
place at the interface. The proximity of an interface might disturb the stability
force balance of the protein, resulting in unfolding of the protein. The unfolding
of proteins upon adsorption is entropically favored (Dill, 1990; Norde, 1986) and
might be the driving force for adsorption where the contact between the hy-
drophobic domains and the aqueous environment are minimized by proper ori-
entation of the molecule (Haynes and Norde, 1994).
tion. Due to this fact, many studies referenced below are concerned with the de-
gree of removal, or elution, of adsorbed protein by surfactant. We have, however,
included these data in this review and tried to evaluate them in connection with
the sparse data that allow understanding on a molecular level. Even if the data
mainly refer to solid surfaces, the basic principles are also valid at liquid inter-
faces such as those of the emulsion droplet. Since the process of surfactant inter-
action with proteins at interfaces is determined by the surfactant/protein, the
surfactant/surface and protein/surface interactions, the following brief introduc-
tion is intended to provide a background on surfactant association and adsorp-
tion, and on surfactant/protein interactions in solution.
The polarized structure of surfactants derives from the hydrophilic head
group and a hydrophobic, usually hydrocarbon, part that promotes interaction
with each other, other molecules, and surfaces by the hydrophilic head as well
as the hydrophobic moieties. These interactions can be expected to be of elec-
trostatic or polar, steric, and hydrophobic type, and the total interaction is
often a sum of or a balance between two or more of these forces. As a conse-
quence of the strong tendency for the hydrophobic chains to avoid contact with
water, self-association will take place both in solution and at interfaces (see
Section 5.4.4). The monomer concentration in solution will be strongly depen-
dent on the association pattern and thus have a pronounced effect on the inter-
facial behavior (Lindman and Wennerstrom, 1980; Israelachvili, 1992).
The general features of surfactant adsorption are
These aspects of surfactant adsorption are illustrated in Figure 5.1 for the
adsorption of dodecyl trimethyl ammonium bromide (DTAB) onto methylated
silica and in Figure 5.2, where a schematic adsorption isotherm is shown.
There is a vast literature concerning the association of surfactants at
solid/aqueous interfaces (Somasundaran and Kunjappu, 1989; Pashley and
100 Food Emulsifiers and Their Applications
~ 0.05
"'su t
6
'bn 0.04 ~ooooosoo8~
§ 0.03
~
0
0
s
«<
§
0
"a 0.02
§
\
Q)
-€
0
"a
Ul 0.01
<:t;
_Q
0.00
0 1800 3600
Time (s)
Figure 5.1 The adsorption of DTAB (2 X erne in phosphate-buffered saline pH 7, I=
0.17) onto methylated silica; rinsing took place after 1800 seconds of adsorption
(Wahlgren, 1992). (Reprinted with kind permission of the American Chemical Society,
Washington, DC)
r iv
ii
c
Figure 5.2 A schematic illustration of an isotherm for adsorption of ionic surfactants
to hydrophilic surfaces (Somasundaran and Kunjappu, 1989). Different stages of the
isotherm are labeled (I) to (IV), and schematic drawings of possible structures that exist
in these regions are presented in Figure 5.3. (Reprinted with kind permission of the
American Chemical Society, Washington, DC.)
length of 6 carbon atoms. However, they found, as expected, that the chain
length did influence the surfactant concentration at which the onset of protein
removal was initiated. The trend was similar to the one observed for the onset
of other cooperative binding events (e.g., micelle formation).
Rapoza and Horhett (1990h) found that surfactants with large head groups
such as Tween 20 gave rise to lower fibrinogen elutahility levels than other sur-
factants at polyethylene surfaces. Welin-Klintstrom et al. (1993) found that the
elutahility of fibrinogen adsorbed at surfaces with a wettahility gradient de-
creased with the bulkiness of the hydrophobic part of the surfactant. In this con-
nection it was also found that nonionics showed an increased removal of
fibrinogen into the more hydrophilic region of the gradient surface when the
cloud point (phase separation temperature) was approached (Wahlgren et al.,
1995). These general observations of removal efficiency are in line with the find-
ings of Backstrom and coworkers (Backstrom, 1987; Lindman et al., 1988), who
studied the removal of fat by different surfactants and found a maximum at con-
ditions corresponding to an optimum in the packing of surfactant molecules at a
flat interface and those of Malmsten and Lindman (1989), who investigated,
among other variables, the effect of temperature on cleaning of hard surfaces.
Thus, it may be concluded that at high surfactant concentrations, head group
effects are, as expected, most pronounced at hydrophilic surfaces hut less impor-
tant at hydrophobic ones. In addition, it appears that principles for detergency in
general, involving the packing efficiency of molecules at interfaces, are applica-
ble to qualitatively describe the removal of proteins from the surface.
toglobulin and lysozyme decreased roughly in the order silica > chromium ox-
ide > nickel oxide (Wahlgren and Arnebrant, 1991). The similarity between
the behavior of the two oppositely charged surfactants indicates that the re-
movability of protein in these cases mainly reflects the binding mode of the
protein to the surface.
Elwing et al. (1989, 1990) studied the surfactant elutability of proteins ad-
sorbed to a surface containing a gradient in hydrophobicity and found large
differences in the amounts removed from the hydrophilic and hydrophobic
ends. In the case of a nonionic surfactant (Tween 20), the elutability was
largest at the midpoint of the gradient, which can be attributed to enhanced
conformational changes of the adsorbed protein at the hydrophobic end, in
combination with a lower efficiency of removal by nonionics at hydrophilic
surfaces. At hydrophobic surfaces the removal is generally high (Elwing et al.,
1989; Wannerberger et al., 1994). However, this may not be considered as evi-
dence for weak binding of the proteins to the surface, but rather as an indica-
tion of the strong interaction between the surfactants and surface.
Of course, the surfactant might bind to the surface and/or to the adsorbed
protein without any net effect on the amount of protein adsorbed. A partial re-
moval according to mechanisms (1) or (2) as illustrated in Figure 5.4 may be
the most common observation and has previously been suggested as one indi-
cation of the presence of multiple adsorption states of the protein (Horbett and
Brash, 1987).
It is important to note that surfactant can remove proteins without binding to
the surface [case (1), left side of Figure 5.4]. This could be described as a solubi-
106 Food Emulsifiers and Their Applications
• ~·
Jones and Wilkinson, 1976; Coke et al., 1990: Frapin et al., 1990;
O'Neill and Kinsella, 1987; Kresheck et al., 1977; Clark et al., 1992;
Creamer, 1995), serum albumin (Tanford, 1980; Nozaki et al., 1974;
Brown, 1984; Ericsson and Hegg, 1985), and specific lipid-binding pro-
teins such as puroindoline from wheat (Wilde et al., 1993).
2. Cooperative adsorption of the surfactant to the protein without gross con-
formational changes.
3. Cooperative binding to the protein followed by conformational changes
(Few et al., 1955; Subramanian et al., 1986; Su and Jirgensons, 1977;
Nelson, 1971).
These different interactions can occur in the same system upon increasing the
surfactant concentration. The conformational changes that occur in case (3)
can involve changes in secondary structure (Nozaki et al., 1974; Subramanian
et al., 1986; Su and Jirgensons, 1977). Several models for the protein/surfac-
tant complexes have been suggested, e.g.: rigid rod (Reynolds and Tanford,
1970), pearl and necklace ( Shirahama et al., 1974), and flexible helix model
(Lundahl, 1986). In the cooperative region (2 and 3), above the critical associ-
ation concentration (cac ), the interaction is mainly one of hydrophobic charac-
ter (Tanford, 1980; Subramanian et al., 1977; Nelson, 1970).
The adsorption from surfactant/protein mixtures to hydrophobic solid sur-
faces is to some extent analogous to the adsorption at air/water or oiVwater in-
terfaces, which have been the subject of frequent studies (Courthaudon et al.,
1991a, b; Clark et al., 1994). Competitive adsorption between proteins and
surfactants at these interfaces has recently been reviewed by Dickinson and
Woskett (1989). They conclude that, as expected, small surface-active compo-
nents above a certain critical concentration will dominate over proteins at
these interfaces, since such components normally have higher surface activity
(superiority in lowering interfacial tension).
Experimentally it is observed that the presence of surfactants in protein so-
lutions may influence the amount of proteins adsorbed to solid surfaces in
three different ways (Wahlgren 1992; Wahlgren and Arnebrant, 1991, 1992;
Wahlgren et al., 1993, 1995):
The complete lack of adsorption in case (1) could be explained by complex for-
mation with a surfactant resulting in an entity that has no attraction to the surface
or direct/preferred surfactant adsorption, due to their higher surface activity and
diffusivity, unpending adsorption of proteins, or protein/surfactant complexes. In
cases (2) and (3), the formation of complexes in solution leads to a decrease or in-
crease in the amounts of protein adsorbed, respectively. The presence of surfactant
influences the total amount of protein adsorbed by steric effects or changes in the
electrostatic interaction between complexes as opposed to native protein. In addi-
tion, the complex may adsorb in a different orientation than the pure protein, re-
sulting in a positive or negative effect on the adsorbed amount.
Due to the different shapes of the adsorption isotherms of surfactants and pro-
teins, the interaction with the interface is of course, as for protein mixtures,
strongly dependent on the relative concentration of the components. The special
character of the surfactant adsorption isotherms featuring the sharp increase in ad-
sorbed amount in the range of their critical association concentration will influence
these events in a very pronounced way. Studies regarding these surfactant/protein
"Vroman effects" have been reported; for example, adsorption of fibrinogen from
mixtures containing Triton X-100 passes through a maximum (Slack and Horbett,
1988). The adsorption from ~-lactoglobulin/SDS mixtures at different degrees of
dilution was studied by Wahlgren and Amebrant (1992) (see Figure 5.5). At con-
centrations above the erne for the surfactant, the amount adsorbed corresponded to
a layer of pure surfactant and was found to increase after rinsing. At lower concen-
trations, the adsorbate prior to rinsing appeared to be a mixture of protein and sur-
factant, and the total amount adsorbed passes through a maximum. The
composition of the adsorbate after rinsing is most likely pure ~-lactoglobulin, since
interactions between the surface or protein and SDS are reversible.
At high degrees of dilution of the mixture, the absence of surfactant adsorp-
tion to the methylated silica, the nonreversible adsorption of ~-lactoglobulin,
and the observed partial desorption of the adsorbate from the mixture imply
that some SDS molecules are bound to ~-lactoglobulin molecules with a higher
affinity than to the surface (see Figure 5.5). The amount of protein adsorbed is
larger, even after rinsing, than for adsorption from pure ~-lactoglobulin solu-
tions, and it can be concluded that SDS binding in this case facilitates the ad-
sorption of protein.
0.18
..-...: .-- .
0 ......-: --
0.12
0.06
1000 100 10
Degree of dilution
Figure 5.5 The amounts adsorbed to a methylated silica surlace as a function of de-
gree of dilution for a mixture of ~-lactoglobulin and SDS (0.2 w/w), in phosphate
buffered saline pH 7, I= 0 .17. The figure shows the adsorbed amount (!lg/cm2) after 30
minutes of adsorption (e) and 30 minutes after rinsing(+). In addition, the figure shows
the adsorption of pure ~-lactoglobulin, after 30 minutes of adsorption (•) and 30 min-
utes after rinsing (x). Finally, the adsorption isotherm of SDS is inserted (0). (From
Wahlgren and Arnebrant, 1992.)
5.3.2.1 Foam and Emulsion Film Stabilization. Thin films are stabilized by
two distinct mechanisms; the one that dominates is dependent upon the molec-
ular composition of the interface (Clark, 1995). Low molecular weight surfac-
tants such as food emulsifiers or polar lipids congregate at the interface and
form a fluid-adsorbed layer at temperatures above their transition temperature
(see Figure 5.6a). When a surfactant-stabilized thin film is stretched, local
thinning can occur in the thin film. This is accompanied by the generation of a
surface-tension gradient across the locally thin region. Surface tension is high-
est at the thinnest point of the stretched film, due to the local decrease in the
surface concentration of emulsifier in the region of the stretch. Equilibrium
surface tension is restored by adsorption of surfactant from the interlamellar
liquid, which is of very limited volume in a drained thin film. This process is
called the "Gibbs effect." Alternatively, migration of the surfactant by lateral
diffusion in the adsorbed layer toward the region of highest surface tension
Protein/Emulsifer Interactions 111
may also occur (Clark et al. 1990a). Here, the surfactant drags interlamellar
liquid associated with the surfactant head group into the thin region of the film
and contributes to the restoration of equilibrium film thickness. This process is
often referred to as the "Marangoni effect" (Ewers and Sutherland, (1952).
In contrast, the adsorbed layer in protein-stabilized thin films is much
stiffer and often has viscoelastic properties (Castle et al., 1987). These derive
from the protein/protein interactions that form in the adsorbed layer (see
Figure 5.6b). These interactions result in the formation of a gel-like adsorbed
layer, recently referred to as a "protein-skin" (Prins et al., 1995), in which lat-
eral diffusion of molecules in the adsorbed layer is inhibited (Clark et al.,
l990b). Multilayer formation can also occur and serves to further mechani-
cally strengthen the adsorbed layer (Coke et al., 1990). When pure protein
h igh mobility
~ Film sttetchiog ~
~
diffusion~
rapid ~Prot
~ cin dcfonnntion
Pilm stretching
No Protein
Slow diffusion
J defonnation
P'lm rui l r e.
i
-
-
Figure 5.6 Schematic diagram showing the possible mechanisms of thin-film stabi-
lization. (a) The Marangoni mechanism in surfactant films; (b) the viscoelastic mecha-
nism in protein-stabilized films; (c) instability in mixed component films. The thin films
are shown in cross section and the aqueous interlamellar phase is shaded. (Reprinted
with kind permission of the Royal Society of Chemistry, London.)
112 Food Emulsifiers and Their Applications
films are stretched, the change in interfacial area is dissipated across the film,
due to the cohesive nature of the adsorbed protein layer and possibly the de-
formability of the adsorbed protein molecules.
Thin-film instability can result in systems that contain mixtures of proteins
and low molecular weight surfactants (Coke et al. 1990; Clark et al., 1991a;
Sarker et al., 1995), as is the case in many foods. The origin of this instability
rests in the incompatibility of the two stabilization mechanisms: the Marangoni
mechanism relying on lateral diffusion, and the viscoelastic mechanism on im-
mobilization of the protein molecules that constitute the adsorbed layer. One
can speculate that in a mixed system, competitive adsorption of low molecular
weight surfactant could weaken or interfere with the formation of protein/pro-
tein interactions in the adsorbed layer and destroy the integrity and viscoelas-
tic properties of the adsorbed layer (see Figure 5.6c). This could be a
progressive process, with the presence of small quantities of adsorbed surfac-
tant initially introducing faults or weaknesses in the protein film. Adsorption
of more surfactant could induce the formation of protein "islands" in the ad-
sorbed layer. These structures could be capable of slow lateral diffusion but
would be too large to participate in Marangoni-type stabilization. Indeed, they
could impede surfactant migration in the adsorbed layer. Adsorption of pro-
gressively more surfactant would reduce the size of the protein aggregates still
further until the adsorbed protein was in its monomeric form. Ultimately, all
the protein would be displaced from the interface by the surfactant.
Two types of interaction are shown in the schematic diagram of the mixed sys-
tem. First, there is an interactive process associated with the coadsorption or
competitive adsorption of the two different species at the interface. Second, many
of the functional proteins used in food production have physiological transport ac-
tivities and therefore possess binding sites, which may allow the formation of
complexes with surfactants. Let us consider each of these processes in tum. The
transition in adsorbed-layer structure at the air/water and oil/water interface
caused by competitive adsorption between protein and emulsifiers has been stud-
ied in detail. Oscillatory surface shear (Kragel et al., 1995) and dilation (Clark et
al., 1993: Sarker et al., 1995) measurements have been carried out at the air/wa-
ter interface and show an emulsifier-induced reduction in surface shear viscosity
and surface dilational elasticity at critical emulsifier/protein ratios. This is consis-
tent with the cartoon depicted in Figure 5.6, where at a specific molar ratio the
emulsifier will break down protein/protein interactions in the adsorbed layer, re-
sulting in a reduction in surface shear viscosity and dilational elasticity. Similar
Protein/Emulsifer Interactions 113
observations have been made at the oil/water interlace under conditions of con-
stant shear in experiments where both components were present in solution when
the interlace was formed (Courthaudon et al., 1991) and when the competing sur-
factant was added subsequent to formation of the protein-adsorbed layer (Chen
and Dickinson, 1995). Direct measurements of changes in adsorbed-layer rheo-
logical properties in foam and emulsion films is possible using the fluorescence
recovery after photobleaching (FRAP) technique (Clark et al., 1990a; Clark,
1995). This method relies on the positioning of a fluorescent reporter molecule in
the adsorbed layer either by covalent labeling of the protein of interest with a flu-
orescent moiety such as fluorescein isothiocyanate (FITC) or by use of an amphi-
pathic fluorescent molecule [e.g., 5-N-(octadecanoyl)amino fluorescein, ODAF] at
low (submicromolar) concentrations that are insufficient to alter adsorbed-layer
structure. FRAP allows direct measurement of the lateral diffusion coefficient of
the fluorophore in the adsorbed layer of the thin film. The presence of an exten-
sive network of protein/protein interactions in the adsorbed layers arrests lateral
diffusion of the probe molecule, as is the case in all films stabilized by protein
alone (Clark et al., 1990b). This situation persists in the presence of emulsifier
until a critical ratio of emulsifier/protein is achieved. This is marked by the onset
of surlace lateral diffusion in the adsorbed layer. The technique serves as a useful
means of (i) comparing the resistance of different proteins to emulsifier-induced
adsorbed layer disruption, (ii) evaluating the effectiveness of protein modification
strategies at improving the resistance of proteins to competitive displacement,
and (iii) investigating the usefulness of natural food ingredients as crosslinkers of
proteins in the adsorbed layer.
5.3.2.2 Specific Binding of Proteins and Emulsifiers. Let us now consider the
effects of specific binding of emulsifiers by food proteins on adsorbed-layer
properties at the air/water and oil/water interlace. There are many examples of
proteins that possess binding activity, including bovine serum albumin and ~
lactoglobulin (see Section 5.3.1.2). Investigation of the binding properties of
these proteins has been generally confined to studies in bulk solution. For ex-
ample, the presence of a fluorescent tryptophan residue in the hydrophobic
cleft of ~-lactoglobulin (Papiz et al., 1986) has facilitated the study of emulsi-
fier binding by fluorescence titration. Subsequent analysis of binding by con-
ventional methods such as that of Scatchard (1949) allows determination of the
dissociation constant (Kd) of the complex formed. Typical examples of KJs for
~-lactoglobulin are shown in Table 5.1. The effect of complex formation can
114 Food Emulsifiers and Their Applications
Dissociation
Emulsifier constant Reference
The general features described earlier are evident with a comparatively low
concentration of protein causing a significant reduction in y. In the absence of
protein, yreduces gradually with increasing Tween 20 concentration. The gradi-
ent of the reduction in surface tension reduces at higher Tween 20 concentra-
tions (> 30 J1M) but doesn't become completely flat due to failure to attain
equilibrium y, possibly due to the presence of a mixture of surface-active species
in the Tween-20 sample. In contrast, the curve in the presence of protein main-
tains a relatively steady surface-tension value of about 50 mN/m up to Tween-20
concentrations of 25 j.iM due to adsorption of the protein. This means that the
curve for the sample containing protein crosses that of Tween 20 alone. This is
strong evidence for complex formation between the two components, since the
curves cross due to a reduction in the concentration of free emulsifier in solution
due to that which interacts with the protein to form the complex.
Thus, great care must be taken when considering the surface properties of
Protein/Emulsifer Interactions 115
80
70
~
5 60
~
·u.;0
~
Q)
~
Q)
<..>
50
$....
;::l
r:n.
40
30
0 20 40 60 80 100
Figure 5.7 Surface tension isotherm for Tween 20 in the absence (e) and presence
(•) of 0.2 mg/mL ~-lactoglobulin. The data were recorded after 20 minutes adsorption
and are therefore not at equilibrium.
P+E~PE (l)
[P] [E]
(2)
[PE]
where [P101] and [E101] are the total protein and emulsifier m the system.
Substituting (3) and (4) in (2) gives
which can be solved for [PE] and can be used to calculate the relative concen-
trations of the three components. In addition, the binding data, which may
comprise a change in a parameter (e.g., intrinsic fluorescence) caused by for-
mation of the complex may be fitted using this equation, provided there is a
single active binding site and the titration is carried out to saturation.
Alternatively, it is possible to determine the dissociation constant and number
of binding sites from the Scatchard (1949) equation
v
(6)
[E]
where v is the fraction of protein with occupied sites (i.e., [PE]/[P101]). If the
Scatchard plot of v against v/[E] gives a straight line, it indicates the presence
of only one class of binding site. The gradient of this line is -l!Kd, and the in-
tercept on the xaxis gives the number of binding sites, n. If the Scatchard plot
does not give a straight line, then the shape of the curve obtained can be used
to identify if the observed binding is positively or negatively cooperative or the
presence of multiple independent sites. In the former case the Hill equation
can be used to determine the Kd and a cooperativity coefficient (Hill, 1910).
Evidence for the formation of a specific complex in solution by direct mea-
surement or by crossovers in surface-tension/concentration isotherms is not
sufficient to allow the conclusion that intact complex adsorbs directly at the
air/water or oil/water interface. There are few studies that provide convincing
data that support such a hypothesis, and that which is provided often only in-
directly points toward the presence of adsorbed complex at the interface. One
of the best understood systems is that of Tween 20 and ~-lactoglobulin (Coke
Protein/Emulsifer Interactions 117
et al, 1990; Wilde and Clark, 1993), which are known to interact in solution
to form a 1:1 complex characterized by a Kd = 4.6 ~' which has an in-
creased hydrodynamic radius of 5. 7 nm compared to 3.5 nm for P-lactoglobu-
lin alone (Clark et al., 1991b). Detailed measurements of the properties of
foam films formed from a constant concentration of 0.2 mg/mL mixed native
and fluorescein-labeled P-lactoglobulin as a function of increasing Tween-20
concentration (Wilde and Clark, 1993; Clark, 1995) have been reported. This
study revealed that between molar ratios (R) of Tween 20 to P-lactoglobulin of
0.2 to 0.9, there was a progressive increase in the thickness of the foam films
and a corresponding decrease in the amount of adsorbed protein to an inter-
mediate level of approximately 50% of that which was originally adsorbed
(see Figure 5.8). These changes occurred prior to the onset of surface diffu-
sion of the labeled protein as determined by the FRAP technique at R = 0.9
(Coke et al., 1990). The increase in foam-film thickness was unexpected since
protein displacement by the Tween 20 should have reduced the thickness of
the thin film. One persuasive interpretation of the data is that coadsorption or
trapping of the Tween-20/P-lactoglobulin complex in the adsorbed multilay-
ers could account for adsorbed-layer thickening (Clark et al., 1994), since the
complex is known to have an increased hydrodynamic radius (Clark et al.,
1991b). Calculations reveal that this is a distinct possibility, since 16 to 49%
of the P-lactoglobulin present in solution will be in the complexed form in the
R-value range, 0.2 to 0.9. Further confirmation of this explanation comes from
measurements of foam-film thickness at different P-lactoglobulin concentra-
tions but at constant R value. Film thickness data for P-lactoglobulin at 0.2
and 1.0 mg/mL are also shown in Figure 5.8. The completely different thick-
nesses observed at the two protein concentrations can be rationalized in terms
of the amount of Tween-20/P-lactoglobulin complex formed. The sharp reduc-
tion in film thickness observed in the 0.2 mg/mL P-lactoglobulin sample at R
= 0.9 occurs when there is 5.41-LM complex present in the sample. An equiv-
alent amount of complex is present in the 1 mg/mL sample at R = 0.1 and
could account for the immediate reduction in film thickness observed with the
1 mg/mL P-lactoglobulin sample and the complete absence of a film thicken-
ing step at low R values.
Further evidence supporting direct adsorption of the complex formed be-
tween ~-lactoglobulin and Tween 20 comes from dynamic surface-tension
(Ydyn) measurements performed using the overflowing cylinder apparatus
(Clark et al., 1993). The Yclvn isotherm for Tween 20 showed classical sig-
118 Food Emulsifiers and Their Applications
40
--,._,
0
_..,
C l)
~
;::S
0
u 30
OJ
u
...
43
;::S
CIJ
8
-E Cl)
Cl)
20
OJ
];a
u
4
Molar ratio (R)
Figure 5.8 A comparison of foam-film thickness and surface concentration data for ~
lactoglobulin samples as a function of Tween 20 concentration. Surface concentration
of FITC/~-lactoglobulin (0.2 mg/mL) as determined by fluorescence counts (.&); foam-
film thickness for samples containing 0.2 mg/mL (•) and l.O mg/mL (e) ~-lactoglobu
lin. R refers to the molar ratio of Tween 20: ~-lactoglobulin.
face activity of the three components present in solution, Tween 20, ~-lac
toglobulin, and the complex, since 'Ydyn behavior is dominated by the compo-
nent present in the highest concentration. Such observations provide
important evidence for direct adsorption of intact complex, and in this partic-
ular case provides evidence that the observed "induction period" in static y
isotherms could be due to the formation of protein islands (de Feijter and
Benjamins, 1987) and can be abolished if interactions between the adsorbed
protein molecules can be prevented, as is the case with the Tween-20/~-lac
toglobulin complex.
Direct adsorption of complex at the air/water interface also appears to have
importance in functional properties of certain lipid-binding proteins from
wheat called "puroindolines" (Blochet et al., 1991; Wilde et al., 1993). These
proteins show unusual behavior in the presence of lipids that they bind, in
that their foaming properties are generally unaltered and in some cases en-
hanced. A systematic study of the influence of interaction with lysophos-
phatidyl cholines (LPC) of different acyl chain lengths has been completed
(Husband et al., 1995) and has produced persuasive evidence of the impor-
tance of the complex on foaming activity. First, two isoforms of the protein
were investigated, puroindoline-a and -b (the b form has also been referred to
as "friabilin"). Puroindoline-b has a significantly increased Kd for LPC com-
pared to puroindoline-a (i.e., 20-fold weaker binding) and the enhancement
of foaming properties is correspondingly reduced in the b form. Further stud-
ies of the binding of LPC to the a form revealed that the binding became
tighter with increasing acyl chain length, and higher concentrations of the
short-chain-length LPC are needed to achieve optimal foam stability en-
hancement. One interesting observation was that the micellar form of LPC
was the species that bound to the protein. This finding emerged from the ob-
servation that lauryl-LPC showed no interaction with the puroindoline-a until
the levels present exceeded the critical micelle concentration of 400 ~·
This indicates a cooperative binding since it takes place in this concentration
range, and any of the suggested structures for the protein/surfactant com-
plexes described in Section 5.3.1.2, e.g., the pearl and necklace structure,
could be applicable. It seems increasingly likely that the functional proper-
ties of the puroindolines are linked to a role in the transport and spreading of
lipid at the air/water interface analogous to the role of amphipathic lung sur-
factant proteins such as SP-B, with which they have striking structural homol-
ogy (Hawgood and Clements, 1990).
120 Food Emulsifiers and Their Applications
1989) but has also been shown to have high affinity for phospholipids, fatty
acids, and triglycerides (Diaz de Villegas, 1987; Creamer, 1995; Sarker et al.,
1995; Kristensen, 1995). An important application of lipid/protein interactions
was reported by Kurihara and Katsuragi (1993), who found that a lipid/protein
complex, formed between ~-lactoglobulin and phosphatidic acid, could mask
bitter taste. This property was suggested to be specific for phosphatidic acid
since no effect was observed for mixtures of ~-lactoglobulin and phosphatidyl-
choline, triglycerides, and diglycerides. Differential scanning calorimetry mea-
surements confirm the presence of a specific interaction between phosphatidic
acid and ~-lactoglobulin since the presence of distearoylphosphatidic acid
(DSPA) as well as dipalmitoylphosphatidic acid (DPPA) thermally stabilized the
protein, which was not observed when the protein was mixed with phosphatidyl-
choline, phosphatidylethanolamine, or phosphatidylglycerol (Kristensen et al.,
1995). No interaction could be observed if the lipid contained unsaturated fatty
acid residues or if it was mixed in the gel state with the protein. Thus, the results
show that the interactions between ~-lactoglobulin and phospholipids are
strongly dependent on the acyl chain as well as the head group. This is not sim-
ply a question of having a negatively charged head group since no interaction
was observed for phosphatidylglycerol.
The influence of protein structure on lipid/protein interactions has been
demonstrated by Brown et al. (1983), who observed that native ~-lactoglobulin
is unable to bind to phosphatidylcholine vesicles. However, if the protein was
dissolved in a-helix-forming solvent, binding to the phospholipid was ob-
served. Brown suggested that the acyl chains of the lipid interact with the hy-
drophobic interior of the a helix, while the polar head group is likely to
interact with the hydrophilic exterior of the protein (Brown, 1984). The par-
tially unfolded proteins formed during food processing may give helix struc-
tures when interacting with the lipids and these lipid/protein complexes can
improve the emulsification process (Brown, 1984; de Wit, 1989).
of human serum albumin, lgG, and fibronectin from human plasma with phos-
pholipids spin-coated onto methylated silica surfaces depends on the phos-
pholipid head group. He found no interaction of proteins with phospholipids
that have no net charge or shielded charges like phosphatidylcholine, phos-
phatidylethanolamine, sphingomyelin, and phosphatidylinositol, whereas in-
teraction was observed with the phospholipid surfaces containing unprotected
charges like phosphatidic acid, diphosphatidylglycerol, and phosphatidylser-
ine. No differences in adsorption behavior were found for spin-coated surfaces
and Langmuir-Blodgett-deposited phospholipid layers.
The consequences of nonspecific interactions with negatively charged di-
oleoylphosphatidylserine (DOPS) bilayzrs observed for fibrinogen and albumin
in contrast to the specific ones observed for prothrombin have been demon-
strated by Corsel et al. (1986). By analyzing their data in terms of intrinsic
binding and transport rate, they found that the initial rate of adsorption of pro-
thrombin was transport-limited, whereas the rate-determining step for the al-
bumin interaction was the binding. The adsorption rate of fibrinogen was either
transport-limited or controlled by the binding, depending on pH and ionic
strength. Since the DOPS bilayer contained biological binding sites for pro-
thrombin, the interaction was completely reversible. However, the interaction
of fibrinogen and albumin with the bilayer probably induced conformational
changes of the proteins and as a consequence the interaction was irreversible.
Kop et al. (1984) found that a high density of DOPS in the bilayer was needed
for the high-affinity binding of prothrombin. Consequently, an introduction of
dioleoylphosphatidylcholine (DOPC) into the bilayer markedly decreased the
affinity for binding.
action between cytochrome c (positive net charge below pH 10) and phospho-
lipids from egg yolk also suggest that it is determined by electrostatics. Their
results show that the limiting pressures for penetration are 20 and 24 mN/m
for phosphatidylcholine and phosphatidylethanolamine, respectively, whereas
penetration into the phosphatidic acid and diphosphatidylglycerol (cardi-
olipin) monolayers occurred up to pressures (< 40 mN/m) close to the col-
lapse pressure of the film. Furthermore, the presence of sodium chloride
decreased the interaction. Later, Kozarac et al. (1988) confirmed the findings
of Quinn and Dawson with their reflection spectroscopy results. Cornell
(1982) observed a specific interaction between ~-lactoglobulin and egg yolk
phosphatidic acid (e-PA) in spread mixed films at low pH (1.3 and 4) where
~-lactoglobulin carries a positive net charge. No interaction was observed in
the neutral pH range or for egg yolk phosphatidyl choline, (e-PC). ~-lac
toglobulin, adsorbing from solution into a spread monolayers of palmitoy-
loleoylphosphatidylcholine (POPC) and palmitoyloleoylphosphatidylglycerol
(POPG), was found to interact with the lipids only in the acid pH range
(Cornell and Patterson, 1989). The highest amounts of ~-lactoglobulin
(Cornell and Patterson, 1989), a-lactalbumin, or BSA (Cornell et al., 1990)
bound to mixed monolayers of POPC and POPG were observed below the iso-
electric point of the protein, when the lipid layer and the proteins carry an op-
posite net charge, whereas less was adsorbed around and almost nothing
above the isoelectric point. The interaction was also found to be reduced in
the presence of calcium as well as if sodium chloride was added (Cornell et
al., 1990). Bos and Nylander (1995) used the film balance to study the incor-
poraffon of ~-lactoglobulin into monolayers of distearoylphosphatidic acid
(DSPA), distearoylphosphatidylcholine (DSPC), and dipalmitoylphosphatidic
acid (DPPA) and some of their results are shown in Figure 5.9. The highest
rate of incorporation was observed for ~-lactoglobulin into the negatively
charged DSPA monolayer. The rate also increases with ionic strength of the
subphase, which probably is due to a decrease of the repulsion within the
phosphatidic acid protein monolayer. The incorporation into the zwitterionic
DSPC monolayers is, as expected, less salt-dependent. The importance of
electrostatic interactions for the incorporation of proteins and peptides in
lipid layers have also been reported for a number of non-food-related systems
like adsorption of SecA (Breukink et al., 1992), actin (Grimard et al., 1993),
IgG (Lu and Wei, 1993), and opioid peptides and opiate drugs (Bourhim et
al., 1993) into phospholipid monolayers.
124 Food Emulsifiers and Their Applications
DSPC
. I , I
''I' 'I
-3 [t DPPA
(j) -3.5
~ t
r
0_0
-4
-4.5
0 5 10 15 20 25 30
ri (mN/m)
BSA into mixed monolayers of POPC and POPG at such high lipid pressure that
they found it unlikely that the proteins could penetrate into a protein layer of
their own at this pressure. Thus, they concluded that the formation of pure pro-
tein patches is unlikely and that portions of the protein is suggested to be inter-
calated in the lipid monolayer. Fluorescence microscopy together with the
surface-film balance technique has also been used to study the structure of
mixed phospholipid cytochrome c and b films (Heckl et al., 1987). It was found
that proteins were located mainly in the fluid membrane phase that coexisted
with solid lipid domains without protein. The penetration in the lipid monolayer
was reduced with increasing pressure. Cytochrome c (positively charged) was
found to interact with dimyristoylphosphatidic acid (DMPA) mono layers but not
with dipalmitoylphosphatidylcholine (DPPC) layers, from which it was con-
cluded that the interaction was largely of electrostatic nature. The effect of dif-
ferent proteins on the organization of lipid monolayer has been demonstrated by
Aynie et al. (1992). Their results indicate that the IX8ccasein in contrast to ~-lac
toglobulin and ~-casein, probably due to the stronger lipid/protein interaction,
induce an ordering of a monolayer of nitroxide fatty acids on the surface of an
emulsion droplet.
5.4.4.1 Polar Lipid Phase Behavior. The polar lipids, having large charged or
uncharged polar groups, have an amphiphilic nature and will thus associate in
Protein/Emulsifer Interactions 127
aqueous systems. The common feature for this self-association is the formation
of a polar interface, which separates the hydrocarbon and water regions. An
overview of some of the main types of structures is given in Figure 5.10. The
hydrocarbon chains can exist either in a fluid state, as in liquid crystalline
phases, or in a solid state, as in the lipid gel phases (Larsson, 1994).
Polar lipids can be further divided into two classes on the basis of their in-
teraction with water:
The water-soluble polar lipids (e.g., ionized fatty acids, bile salts, and syn-
thetic surfactants, charged or uncharged) have monomeric solubility in the mil-
limolar range and form micelles at higher concentrations. The critical micelle
concentration (erne) is considered to be a narrow concentration range, within
which aggregates start to form by a strong cooperative process (Lindman and
Wennerstrom, 1980). The driving force for micelle formation is the hydrophobic
interaction (see Tanford, 1980). For ionic amphiphiles, erne also depends on the
ionic strength, since addition of salt reduces the electrostatic repulsion between
the charged head groups. At low water content an inverse micellar structure, the
1 2 phase, is formed in which the hydrocarbon chains form the continuous
medium and the aqueous medium is present within the micelles.
A common feature of the two classes of polar lipids is the tendency to form
lyotropic liquid crystalline phases (see Figure 5.10). The main features of the
most commonly found mesophases were determined by Luzatti and coworkers
in the early 1960s by x-ray diffraction [reviewed by Luzatti in 1968 (Luzatti,
1968)]. Later spectroscopy studies have given a considerable number of con-
tributions to the understanding of the dynamic nature of these phases. The
lamellar phase (La) consists of stacked infinite lipid bilayers separated by wa-
ter layers, while the hexagonal phases consist of infinite cylinders, having ei-
ther a hydrocarbon core (H 1) or a water core (Hu)· The cubic phases have been
shown to exist in a number of lipid systems (Fontell, 1990) and their possible
role in biological systems has been discussed by Larsson (1989). Since the cu-
bic phases are isotropic and highly viscoelastic, they can be hard to analyze
and to handle. Different structures of the cubic phases have been suggested,
which depend on the particular lipid system (Larsson, 1989; Lindblom and
128 Food Emulsifiers and Their Applications
Reversed micelles
(L2)
(Solid)
(Melt) <-Cubic
z Reversed hexagonal
0
i= (HII)
<1::
a:
f-
z "Water-in-oil"
w
u <-Cubic (Q)
z
0
u
f-
z I"Mirror plane" I Lamellar
<1::
f- (La)
u
<1::
LL.
a:
::l <-Cubic
(/)
"Oil-in-water"
c
w
(/)
<1::
w Hexagonal
a: (Hi)
u
~
e
<·Cubic
(H20) Micelles
(L1)
Figure 5.10 Commonly formed association structures by polar lipids. Phase transi-
tions can be induced by changes in water content, temperature, or interaction with
other solution components, like proteins. The lamellar liquid crystalline phase (LJ ,
can be regarded as the mirror plane, where the aggregates are of the "oil-in-water" type
on the water-rich side and of the "water-in-oil" type on the water-poor side (Fontell,
1992). On both the water-rich and water-poor sides of the L" there are two possible lo-
cations for cubic phases. Other "intermediate phases" may also occur.
Rilfors, 1989; Fontell, 1992). One type of cubic structure, consisting of rod-
like aggregates, has been proposed by Luzatti et al. (1968). The two polar rod-
like networks formed in this way give two continuous and unconnected
systems of fluid hydrocarbon chains. The other main type of cubic phases, ob-
served for water-insoluble lipids, are bicontinuous and based on curved nonin-
tersecting lipid bilayers, which are organized to form two unconnected
continuous systems of water channels (see Lindblom et al., 1979; Larsson,
1989). If an interface is placed in the gap between the methyl end groups of
Protein!Emulsifer Interactions 129
the lipid, it will form a plane that can be described as a minimal smface
(Andersson et al., 1988; Larsson, 1989). A minimal surface has zero average
curvature at any point on the surface; that is, in all points the surface is as con-
cave as it is convex. A minimal surface exhibiting periodicity, as in the cubic
lipid aqueous phase, is termed an "infinite periodic minimal surface" (IPMS),
and three types of IPMS, with different geometry, have been shown to be im-
portant in lipid systems (Andersson et al., 1988; Larsson, 1989).
With decreasing water content, the phase behavior of the polar lipids often
follows this sequence: hexagonal phase (HJ ~ lamellar phase (LJ for water-
soluble lipids and lamellar phase (LJ ~ reversed hexagonal phase (H1J for
lipids with low water solubility. Cubic liquid crystalline phases (Q) often occur
in between these. Phase transitions can also occur with changes in tempera-
ture; with increasing temperature the sequence of thermal transitions is usu-
ally the same as with decreased water content.
In nature and in many technical applications the aggregates consist of a
mixture of different lipids, which either exist in a homogenous mixture or sep-
arate into domains. As discussed in the review by Raudino (1995), the lateral
distribution in these mixed aggregates is influenced by a number of factors like
ionic strength, presence of polymers/proteins, as well as the composition of the
lipids; thus it is hard to give any general rules to predict when phase separa-
tion will occur.
5.4.4.2 Some Aspects of the Interaction between Proteins and Lipid Structures.
Interactions between proteins and lipid structures are of fundamental importance
in biological membranes, as well as in food emulsions. Our aim is to give some ex-
amples of the effects the interaction can have on the lipid phase behavior as well
as on the protein structure. The same type of molecular forces, as discussed in
previous sections, are responsible for the interaction.
Much of the work on lipid phase transitions induced by proteins and pep-
tides has been focused on intrinsic and peripheral membrane proteins. Many
of these proteins carry a positive net charge at neutral pH, and most biological
membranes are neutral, zwitterionic, or negative under physiological condi-
tions (Sankaram and Marsh, 1993). Binding of these proteins generally takes
place via direct electrostatic interaction between basic residues, like lysine
and arginine, and the opposite charges on the lipids. However, other effects
like hydrophobic interaction and steric factors can contribute significantly.
Gramicidin, an uncharged and hydrophobic pentadecapeptide, favors the tran-
130 Food Emulsifiers and Their Applications
It was observed that in the fluid state of pure DMPG and DOPG, the bound
protein existed in the more unfolded form (II), whereas in the gel state of
DMPG the native form (I) prevailed. When DOPG, was mixed with nonionic
(DOG) or zwitterionic (DOPC), the bound fraction of the native form (I) in-
creased with the content of neutral lipids.
The work of Minami et al. (1995), in which incorporation of P-lactoglobulin
and P-lactalbumin with negative net charges and lysozyme with positive net
charge at neutral pH, into a sphingomyelin/palmitate gel phase, was studied
found no correlation with the protein net charge. Instead, the amount of pro-
tein, which could be dissolved in the thin aqueous layer of the gel phase was
Protein/Emulsifer Interactions 131
decreased in the order lactate oxidase > creatinine deiminase > glucose oxi-
dase > urease, and corresponded to the order of increasing molecular weight.
Further investigations have shown that the introduction oflecithin in the cubic
phase can increase the stability of entrapped glucose oxidase (Nylander et al.,
1995). The mobility of the entrapped enzyme is limited in the cubic phase
compared to bulk solution (1995), whereas the diffusion measurements of
Mattisson et al. (1995) show that a typical substrate molecule, like glucose,
can move relatively freely into the cubic phase. These studies point to an im-
portant potential application of cubic lipid phases as a flexible and biocompat-
ible matrix for the immobilization of enzymes for synthesis as well as for
analytical applications. The cubic monoglyceride phases also have the ability
to solubilize lipophilic proteins as well as relatively large amounts of mem-
brane lipids. Larsson and Lindblom (1982) have reported the solubilization of
a-gliadin, which is a hydrophobic wheat protein fraction, into a cubic 1-
monoolein aqueous phase, with the same characteristics as a binary cubic 1-
monoolein phase. In this case, the proteins most probably are dispersed in the
lipid bilayer region of the cubic phase.
The formation of the interfacial layer on the emulsion droplet is strongly linked
to the properties of the aqueous environment as well as to the properties of the
oil phase. An example of the former has been provided by Chen and Dickinson
(1995a-c), who showed that the addition of an anionic surfactant, sodium lau-
ryl sulfate (SLES), can introduce flocculation of a gelatin stabilized oil-in-wa-
ter emulsion at sufficient surfactant concentrations. An increase of the SLES
concentration led to a restahilization of the flocculated emulsion. This could he
related to the hulk behavior, where the anionic surfactant will neutralize the
positively charged gelatin and cause precipitation of the protein. As more sur-
factant is added, the solubility of the protein increases due to binding of sur-
factant that increases the protein net charge, and finally the precipitate is
redissolved. Measurements of the composition of the interfacial layer reveal
(Chen and Dickinson, 1995c) that gelatin is initially replaced by SLES, hut as
the surfactant concentration increases, more gelatin can he accommodated at
the interface due to partial neutralization of the gelatin at the interface. This
will eventually cause flocculation to occur (Chen and Dickinson, 1995a). A
further increase of surfactant concentration leads to a decrease in gelatin sur-
face concentration (Chen and Dickinson, 1995c) and a restabilization of the
emulsion (Chen and Dickinson, 1995a). It is noteworthy that an emulsion sta-
bilized by P-lactoglohulin, which was negatively charged under the prevailing
experimental conditions, did not show any signs of flocculation upon addition
of SLES, although a complex was formed in the hulk solution.
Several examples of how the properties of the oil phase composition can af-
fect the structure of the adsorbed layer of protein on the emulsion droplet, and
hence the stability of the emulsion, have been studied. For instance, the work
of Leaver and Dagleish (1992) on the structure of adsorbed layers of ~-casein
on emulsion droplets, where it was found that the cleavage of the protein on
the oil-droplet surface by trypsin gave different products depending on
whether a triglyceride oil or tetradecane, was used. This demonstrates that the
structure of the adsorbed layer depends on the composition of the oil.
Furthermore, it was shown by Heertje et al. (1990) that if a monoglyceride was
dissolved in the oil phase, the displacement of caseinate from the oil/water in-
terface was correlated to the interfacial concentration of monoglyceride. They
also found that at high monoglyceride concentration in the oil phase, the
amount of saturated (monostearoylglycerol) lipid adsorbed at the interface was
Protein/Emulsifer Interactions 135
larger than that of the unsaturated (monooleoylglycerol) one, which also led to
more extensive displacement of caseinate.
We have in the foregoing section mainly been concerned with the binding of
emulsifiers to proteins, but one can also think of cases where there is a repul-
sion between the lipid and head group and the protein. In this way the struc-
ture of the adsorbed protein layer, and hence the properties of the emulsion,
can be changed. This has been demonstrated by Brookbanks et al. (1993), who
found that the thickness of adsorbed layers of milk proteins was significantly
greater when the protein was adsorbed onto negatively charged phosphatidyl-
glycerol (PG) liposomes. The more extended structure was attributed to charge
repulsion between the negatively charged lipid surface and the negatively
charged surface domains of the protein.
5.5.3 Conclusion
The interaction between emulsifiers and proteins is to a larger extent driven by
electrostatic or hydrophobic interactions, or in many cases it is a combination
of the two. Thus, it is commonly observed that ionic emulsifiers interact more
strongly with proteins than nonionic ones. For emulsifiers with low water solu-
bility, e.g., polar lipids, the interaction with proteins is largely dependent on
the phase structure upon addition. The binding can, depending on the type of
emulsifier, lead to stabilization of the protein structure at low-surfactant-to-
protein ratios. However, an increase in surlactant concentration can induce
unfolding of the protein and in some cases precipitation of the protein.
We have seen that the stability of emulsions and foams is determined by in-
terfacial processes, which are affected by the properties of the interface as well
as the interactions occurring in bulk solution.
When no emulsifier/protein interactions are present, the composition of the
interfacial film is determined by only the surface activity and concentration of
the components. In the case of reversibility the "race for the interface" is won
by the most surface-active and/or abundant molecule, and in the case of irre-
versibility the transport rate might also play a role. In this context it has to be
born in mind that proteins can change their conformations (sometimes in a
time-dependent way) at the interface. This may lead to a strong interaction be-
tween the protein and the surface, which can hamper the displacement of a
protein by more surface-active emulsifiers.
The presence of protein/emulsifier interactions can have pronounced im-
pact on the interfacial behavior of the components. In cases where the emulsi-
136 Food Emulsifiers and Their Applications
Acknowledgement
Thomas Arnebrant thanks the Swedish Research Council for Engineering
Sciences (TFR) and Martin Bos acknowledges the EU Human Capital and
Mobility program (ERBCHRXT930322) for their financial support. Tommy
Nylander acknowledges the Swedish Council for Forestry and Agricultural
Research. David Clark acknowledges the support of the Biotechnology and
Biological Sciences Research Council.
References
Ananthapadmanabhan, K.P., (1993). Protein-surfactant interactions, in Interactions of
Surfactants with Polymers and Proteins (eds. K.P. Ananthapadmanabhan and E.D.
Goddard), CRC Press, Boca Raton, FL, pp. 319-366.
Andersson, S., et al. (1988). Minimal surfaces and structures: From inorganic and
metal crystals to cell membranes and biopolymers, Chem. Rev., 88, 221-242.
Arai, T., Norde, W. (1990a). The behaviour of some model proteins at the solid/liquid
interfaces. I. Adsorption from single protein solutions, Colloids and Surfaces, 51,
l-16.
- - - (1990b). The behavior of some model proteins at the solid/liquid interfaces. II.
Sequential and competitive adsorption, Colloids and Surfaces, 51, 17-28.
Arnebrant, T., et al. (1989). An ellipsometry study of ionic surfactant adsorption on
chromium surfaces,]. Colloid Interface Sci., 128, 303-312.
Aynie, S., et al. (1992). Interactions between lipids and milk proteins in emulsion, ].
Food Sci., 57(4), 883-887.
Backstrom, K. (1987). Removal of Triglyceride and Protein Films from Hard Surfaces
by Aqueous Surfactant Solutions, an Ellipsometry Study, PhD Thesis, University of
Lund, Sweden.
Blochet, J.E., et al. (1991). Amphiphilic proteins from wheat flour: Specific extraction,
structure and lipid binding properties, in Gluten Proteins 1990 (eds. W. Bushuk and
R. Tkachuk), A.A.C.C., St Paul, MN, pp. 314-325.
Blomberg, E., (1993). Surface Force Studies of Adsorbed Proteins, PhD Thesis, Royal
Institute of Technology, Sweden.
Bohnert, J.L., Horbett, T.A. (1986). Changes in adsorbed fibrinogen and albumin inter-
action with polymers indicated by decreases in detergent elutability, J Colloid
Interface Sci., Ill, 363-377.
Bos, M.A., Kleijn, J.M. (1995a). Determination of the orientation of adsorbed fluo-
rophores using TIRF. I. Theory, Biophys. ]., 68, 2566-2672.
138 Food Emulsifiers and Their Applications
- - - et al. (1992). The interaction of sucrose esters with ~-lactoglobulin and ~-ca
sein from bovine milk, Food Hydrocolloids, 6, 173-186.
- - - et al. (1993). Surface dilational behavior of aqueous solutions of ~-lactoglobu
lin and Tween 20, in Food Colloids and Polymers: Structure and Dynamics (eds. E.
Dickinson and P. Walstra), RSC Special Publication 113, pp 354-364.
- - - e t al. (1994). Differences in the structure and dynamics of the adsorbed layers
in protein stabilised model foams and emulsions, Royal Soc. Chem. Faraday
Discussion, 98, 253-262.
Coke, M., et al. (1990). The influence of surface composition and molecular diffusion
on the stability of foams formed from protein detergent mixtures, ]. Colloid Interface
Sci., 138, 489-504.
Cornell, D.G. (1982). Lipid-protein interactions in monolayers: Egg yolk phosphatidic
acid and ~-lactoglobulin,]. Colloid Interface Sci., 88, 536-545.
- - - , Caroll, R.J. (1985). Miscibility in lipid-protein monolayers,] Colloid Interface
Sci., 108, 226-233.
---,Patterson D.L. (1989). Interaction of phospholipids in monolayers with ~-lac
toglobulin adsorbed from solution,]. Agric. Food Chem., 37, 1455-1459.
- - - et al. (1990). The interaction of phospholipids in monolayers with bovine serum
albumin and a-lactalbumin adsorbed from solution, ]. Colloid Interface Sci., 140,
428-435.
Corse!, J. W., et al. (1986). The role of intrinsic binding rate and transport rate in the ad-
sorption of prothrombin, albumin and fibrinogen to phospholipid bilayers, ]. Colloid
Interface Sci., Ill, 544-554.
Courthaudon, J.-L., et al. (l99la). Competitive adsorption of ~-casein and nonionic
surfactants in oil-in-water emulsions,]. Colloid Interface Sci., 145, 390-395.
- - - et al. (l99lb). Competitive adsorption of ~-lactoglobulin and Tween 20 at the
oil-water interface, Colloids Surf, 56, 293-300.
Creamer, L.K. (1995). Effect of sodium dodecyl sulphate and palmitic acid on the equi-
librium unfolding of bovine ~-lactoglobulin, Biochemistry, 34, 7170-7176.
Creighton, T.E. (1990). Protein folding, Biochem. ]., 270, l-16.
Diaz de Villegas, M.C., et al. (1987), Lipid binding by ~-lactoglobulin of cow milk,
Milchwissenschafi, 42, 357-358.
Dickinson, E.D. (1988). In Food Structure: Its Creation and Evaluation (eds. J.M.
Blanshard and J.R. Mitchell), Butterworths, London, p. 41.
- - - , Stainsby, G. (eds.) (1982). Colloids in Food, Elsevier Applied Science,
London.
- - - , Woskett, C.M. (1989). Competitive adsorption between proteins and small
molecular surfactants in food emulsions, in Food and Colloids (eds. R.D. Bee, P.
Richmond, and J. Mingins), Royal Society of Chemistry, London, pp. 74-96.
Dill, K. A. (1990). Dominant forces in protein folding, Biochemistry, 29, 7133-7155.
Eidem, T., Speiser, P. (19~9). Acta Pharm. Techn., 35, 109.
Elliot, J.R., et al. (1983). Biochim. Biophys. Acta, 464, 127-141.
Elwing, H., Colander, C.-G. (1990). Protein and detergent interaction phenomena on
solid surfaces with gradients in chemical composition, Adv. Colloid Interface Sci.,
32,317-339.
- - - et al. (1989). Desorption of fibrinogen and y-globulin from solid surfaces in-
duced by nonionic detergent,]. Colloid Interface Sci., 128, 296-300.
140 Food Emulsifiers and Their Applications
Ericsson, B. (1986). Interactions between Globular Proteins and Polar Lipids, PhD
Thesis, University of Lund, Sweden.
- - - , Hegg, P.-O. (1985). Surface behavior of adsorbed films from protein-am-
phiphile mixtures, Prog. Colloid & Polymer Sci., 70, 92-95.
- - - et al. (1983). A cubic protein-monoolein-water phase, Biochim. Biophys. Acta,
729, 23-27.
Ertel, S.I., et al. (1991). The adsorption and elutability of albumin, lgG and fibronectin
on radiofrequency plasma deposited polystyrene, ]. Colloid Inteiface Sci., 147,
433-442.
Ewers,W.E., Sutherland, K.L. (1952). The role of surface transport in the stability and
breakdown of foams, Aust. ]. Sci. Res. Ser. AS, 697-710.
De Feijter, J.A., Benjamins, J. (1987). Adsorption kinetics of proteins at the airwater
interface, in Food Emulsions and Foams (ed. E. Dickinson), Royal Society of
Chemistry, London.
Few, A.V., et al. (1955). The interaction between bovine plasma albumin and dode-
cyltrimethylammonium bromide, Biochim. Biophys. Acta, 18, 136-137.
Fidelio, G.D., et al. (1986). Interaction of melittin with glycosphingolipids and phos-
pholipids in mixed monolayers at different temperatures. Effect of the lipid physical
state, Biochem. Biophys. Acta, 862, 49-56.
Fontell, K. (1990). Cubic phases in surfactant and surfactant-like lipid systems,
Colloid Polym. Sci., 268, 264-285.
- - - (1992). Some aspects on the cubic phases in surfactant and surfactant-like
lipid systems, Adv. Colloid lnteiface Sci., 41, 127-147.
Frapin, D., et al. (1993). Probing the fatty acid binding site of 13-lactoglobulin, ]. Prot.
Chem., 12,443-448.
Fraser, P.E., et al. (1989). Bilayer-stabilising properties of myelin basic protein in di-
oleoylphosphatdiylethanolamine systems, Biochem. Biophys. Acta, 983, 23-29.
Gargouri, Y., et al. (1984a). Studies on the inhibition of pancreatic and microbial li-
pases by soybean protein,]. Lipid Res., 25, 1214-1221.
- - - et al. (1984b). Inhibition of pancreatic and microbial lipases by proteins,
Biochim. Biophys. Acta, 795, 326-331. •
- - - et al. (1985). Inhibition of lipases by proteins. A kinetic study with dicaprin
monolayers, ]. Biol. Chem., 269, 2268-2273.
- - - et al. (1987). Human gastric lipase. A kinetic study with dicaprin monolayers,
Eur. ]. Biochem., 169, 125-129.
- - - et al. (1989). Role of a sulphydryl group in gastric lipases. A binding study us-
ing the monomolecular film technique, Eur. ]. Biochem., 180, 367-371.
Gekko, K., Hasegawa, Y. (1986). Compressibility-structure relationship of globular
proteins, Biochemistry, 25, 6563-6571.
Graham, D.E., Phillips, M.C. (1979). Proteins at liquid interfaces. III. Molecular struc-
tures of adsorbed films,]. Colloid lnteiface Sci., 70, 427-439.
Griffin, M.C.A., et al. (1984). Structural domains of K-casein show different interaction
with dimyristoylphosphatidylcholine monolayers, Chem. Phys. Lip., 36, 91-98.
Grimard, R., et al. (1993). Interaction of actin with positively charged phospholipids: A
monolayer study, Biochem. Biophys. Res. Com., 19(3), 1017-1022.
Gumpen, S., et al. (1979). Thermal stability of fatty acid serum albumin complexes
studied by differential scanning calorimetry, Biochim. Biophys Acta, 574, 189-196.
Protein/Emulsifer Interactions 141
Hawgood, S., Clements, J.A. (1990). Pulmonary surfactant and its apo-protein, ]. Clin.
Invest., 86, l-6.
Haynes, C.A., et al. (1994). Globular proteins at solid/liquid interfaces, Colloids Surf
B, 2, 517-566.
Heck!, W.M., et al. (1987). Interactions of cytochrome b5 and c with phospholipid
monolayers, Biochim. Biophys. Acta, 903, 166--176.
Heertje, 1., et al. (1990). The observation of the displacement of emulsifiers by confocal
scanning laser microscopy, Food Structure, 9, 305-316.
Hegg, P.-O. (1980). Thermal stability of ~-lactoglobulin as a function of pH and the rel-
ative concentration of sodium dodecylsulphate, Acta Agric. Scand., 30, 401-404.
Heimburg, T., et al. (1991). Cytochrome c-lipid interactions studied by resonance
Raman and :lip NMR spectroscopy. Correlation between the conformational changes
of the protein and the lipid layer, Biochemistry, 30, 9084-9089.
Hill, A.V. (1910). A new mathematical treatment of changes in ionic concentration in
muscle and nerve under action of electron currents with a theory as to their mode of
excitation,]. Physiol. 40, 90-224.
Horbett, T.A., Brash, J.L. (1987). Proteins at interfaces: current issues and future
prospects, in Proteins at Interfaces: Physicochemical and Biochemical Studies (eds.
J.L. Brash and T.A. Horbett), American Chemical Society, Washington, DC, pp.
1-37.
Husband, F.A., et al. (1995). A comparison of the foaming and interfacial properties of
two related lipid binding proteins from wheat in the presence of a competing surfac-
tant, in Food Macromolecules and Colloids, (eds. E. Dickinson and D. Lorient),
Royal Society of Chemistry, London, pp. 283-296.
lsraelachvili, J.N., Adams, G.E. (1978). Measurement offorces between two mica sur-
faces in aqueous electrolyte solutions in the range 0-100 nm, ]. Chem. Soc.
FaradaJ Trans. 1, 74, 975-1001.
Jones, M.N., Wilkinson, A. (1976). The interaction between !)-1aetog1obulin and
sodium n-dodecyl sulphate, Biochem. ]., 153, 713-718.
Katona, E., et al. (1978). The application of surface tension measurements to the study
of protein-lipid interactions between a hydrophobic meylin protein (lipophilin) and
lipids,]. Colloid Interface Sci., 67, 108-117.
Kauzmann, W. (1959). Some factors in the interpretation of protein denaturation, Adv.
Protein Chem., 14, 1-63.
Kop, J.M.M., et al. (1984). The adsorption of prothrombin to phospholipid monolayers
quantitated by ellipsometry,]. Biol. Chem., 259(22), 13993-13998.
Kozarac, Z., et al. (1988). Adsorption of cytochrome c to phospholipid monolayers stud-
ied by reflection spectroscopy, FEES, 229(2), 372-376.
Krage!, J., et al. (1995). Protein surfactant interaction in mixed adsorption layers at liq-
uid interfaces, Progress in Colloid and PolJmer Science. In press.
Kresheck, G.C., et al. (1977). Thermometric titration studies of ligand binding to
macromolecules. Sodium dodecyl sulphate to blactoglobulin, ]. PhJs. Chem., 81,
532-537.
Kristensen, A., et al. (1995). Interaction between ~-lactoglobulin and phospholipids in
solution, Submitted to ]. Dairy Sci.
Kurihara, K., Katsuragi, Y. (1993). Specific inhibitor for bitter taste, Nature, 365,
213-214.
142 Food Emulsifiers and Their Applications
Landau, E.M., Luisi, P.L. (1993). Lipidic cubic phases as transparent, rigid matrices for
the direct spectroscopic study of immobilized membrane proteins, ]. Am. Chem.
Soc., ll5, 2102-2106.
Larsson, K. (1989). Cubic lipid-water phases: Structure and biomembrane aspects, ].
Phys. Chem. ,93, 7304-7314.
- - - (1994). Lipids-Molecular Organization, Physical Functions and Technical
Applications, The Oily Press, Dundee, Scotland.
---,Lindblom, G. (1982). Molecular amphiphile bilayers forming a cubic phase in
amphiphile-water systems,]. Disp. Sci. Techno!., 3, 61-66.
Leaver, J., Dagleish, D.G. (1993). Variations in the binding of ~-casein to oil-water in-
terfaces detected by trypsin-catalysed hydrolysis, ]. Colloid Interface Sci., 149,
49-55.
Lindblom, G., et al. (1979). The cubic phase of monoglyceride-water systems.
Arguments for a structure based upon lamellar bilayer units, ]. Am. Chem. Soc.,
101, 5465-5470.
- - - , Rilfors, L. (1989). Cubic phases and isotropic structures formed by membrane
lipids-possible biological relevance, Biochem. Biophys. Acta, 988, 221-256.
Lindman, B., Wennerstrom. H. (1980). Micelles amphiphile aggregation in aqueous so-
lutions, Topics in Current Chemistry, 87, 1-83.
- - - et al. (1988). The cleaning of hard surfaces by surfactant in relation to surfac-
tant molecular packing, ]. Surface Sci. Techno!., 4, 23--41.
Lu, B., Wei, Y. (1993). The interaction of phospholipid in monolayer with dye labeled
IgG adsorbed from soluffon, ]. Colloid Interface Sci., 161, 12{}-124.
Lundahl, P., et al. (1986). A model for ionic and hydrophobic interactions and hydro-
gen-bonding in sodium dodecyl sulphate-protein complexes, Biochim. Biophys.
Acta, 873, 2{}-26.
Luzzati, V. (1968). X-ray diffraction studies of lipid-water systems, in Biological
Membranes (ed. D. Chapman). Academic, New York, pp. 77-123.
- - - et al. (1968). Structure of the cubic phases of lipid-water systems, Nature, 220,
485--488.
Makino, S., et al. (1983). Sucrose mono-esters of fatty acids: Their properties and inter-
action with proteins, Agric. Biol. Chem., 4 7, 319-326.
Malmsten, M. (1994). Ellipsometric studies of protein adsorption at lipid surfaces, ].
Colloid Interface Sci., 168, 247-254.
- - - (1995). Protein adsorption at phospholipid surfaces, ]. Colloid Interface Sci.,
172, 106-ll5.
- - - , Lindman, B. (1989). Ellipsometry studies of cleaning hard surfaces. Relation
to the spontaneous curvature of surfactant monolayer, Langmuir, 5, ll 05-ll11.
Mattisson, C., et al. (1995). Diffusivity measurements by holographic laser interferome-
try in a cubic lipid-water phase, Submitted to Chem. Phys. Lipids.
McGuire, J., et al. (1995a). Structural stability effects on adsorption and dode-
cyltrimethylammonium bromide-mediated elutability of bacteriophage T4 lysozyme
at silica surfaces,]. Colloid Interface Sci., 170, 182-192.
- - - et al. (1995b). Comparative adsorption studies with synthetic structural stabil-
ity and charge mutants of bacteriophage T4lysozyme, in Proteins at Interfaces (eds.
J.L. Brash and T.A. Horbett), ACS Books, Washington, DC, Vol. 602, pp. 55-65.
- - - et al. (1995c). The influence of net charge and charge location on adsorption
Protein/Emulsifer Interactions 143
Wallin, R., et al. (1993). Stabilisation of glucose oxidase by entrapment in a cubic liq-
uid-crystalline phase, Biocatalysis, 8, 73--80.
Walstra, P. (1983). In Encyclopedia of Emulsion Technology (ed. P. Becher), Marcel
Dekker, New York, Vol. 1, p. 57.
Wannerberger, K., et al. (1995). Adsorption onto methylated silica surfaces from li-
pase/surfactant solutions, Colloids and Surfaces B: Biointerfaces. In press.
Watts, A. (ed.) (1993). Protein-Lipid Interactions, New Comprehensive Biochemistry,
Vol. 25, Elsevier, Amsterdam.
Welin-Klintstrom, S., et al. (1993). Surfactant and protein interactions on wettability
gradient surfaces,]. Colloid Interface Sci., 158, 188-194.
Wilde, P.J., Clark, D.C. (1993). The competitive displacement of ~-lactoglobulin by
Tween 20 at the oil-water interface,]. Colloid Interface Sci., 155, 48-54.
- - - , Marion, D. (1993). The influence of competitive adsorption of a lysopalmitoyl
phosphatidylcholine on the functional properties of puroindoline, a lipid binding
protein isolated from wheat flour,]. Agric. Food Chem., 41, 1570-1576.
De Wit, J. N. (1989). Functional properties of whey proteins, in Developments in Dairy
Chemistry-4 (ed. P.F. Fox), Elsevier Applied Science, London, pp. 285-322.
Wangnerud, P. (1994). Adsorption oflonic Surfactants on Charged Solid Surfaces, PhD
Thesis, University of Lund, Sweden.
SIX
Physicochemical Aspects of
an Emulsifier Functionality
Bjorn Bergenstahl
6.1 Introduction
The characteristic property of all emulsifiers is their surface activity. Surface
activity is the ability to form a surface excess at interfaces. The formation of
adsorbed layers at interfaces are displayed in a change of a range of easily ob-
servable and technically important properties.
147
148 Food Emulsifiers and Their Applications
This chapter aims to discuss the principal physical origin of the various func-
tionalities of typical lipid food emulsifiers. Aspects on the functionality under
very different conditions in various foods will be discussed. I will try to show how
we may select emulsifiers on the basis of their fundamental properties. ·
t Several lipid emulsifiers are exceptions and are applied in a hydrated gel form (a crystals).
However, this crystal form resembles the liquid crystalline form in terms of interactions with
both phases and spreadability over the interfaces.
Physicochemical Aspects of an Emulsifier Functionality 149
The solution properties of emulsifiers are determined for the surface activ-
ity of the emulsifiers. In addition, the ability to generate repulsive interactions
is also reflected in the solution properties of emulsifiers.
reversed
micelles
reversed
hexagonal
lamellar phase Melting point
M---
phase
hexagonal
, phase
:micelles
'
'
Krafft
point
Emulsifier
where .ilGmixing is negative when changing from large aggregates to small ag-
gregates (micelles and molecular solutions).
is positive and equal to Ahydrocarbonlwater 'Y hydrocarbon/water· The hy-
ilGhydrophobic
drophobic effect is the driving force for the aggregation and gives the upper
limit of the molecular solubility for amphiphilic molecules (critical micelle
concentration) .
is negative. This term consists mainly of the work released
.ilGpolar group/water
L l I
next aggregate }
where l is the average distance between the polar groups and F(l) is the inter-
action.
The area per molecule in the aggregates is given by the balance between
the interfacial tension of the oil/water interface and of the space needed for the
Physicochemical Aspects of an Emulsifier Functionality 151
polar group itself and the space generated by repulsive interactions between
the emulsifier head groups at the interface.
The area per molecule expands in the series Areversed micelles < Areversed hexagonal
< Alamellar Ahexagonal < Amicelles"
At a specific ratio of water and emulsifier, the system's tendency is to obtain
aggregates as small as possible to maximize the AGmixing and the AGpolar group/water·
The lower limit in aggregate size is given by the increased hydrophobic contact
between the exposed hydrocarbon/water interface.
The interesting result of this exercise is that the area per molecule is to a
large extent a measure of the ability to generate repulsive interactions.
In the solubilization sequence, reversed aggregates ~ lamellar phase ~
hexagonal phase~ micellar solution~ molecular solution, the area per mol-
ecule of the surfactant/water interface increases. Depending on the packing
constraints given by the hydrophobic moiety in the aggregates, the range of the
repulsive interaction on the polar side of the molecule, and the molecular
weight, this process has to proceed more rapidly or more slowly (Israelachvili
et al., 1976, 1977). Hence, the packing constraints of the hydrocarbon chain
are an important link between properties and aggregation.
The ratio of the actual area A, as it is created by the repulsive interactions,
to the theoretical area of a saturated hydrocarbon chain, A0 (23 A2) enforces
different geometries (Israelachvili et al., 1976, 1977) due to the different ratio
of volume to area of different aggregates, as shown in Table 6.2.
The successive solvation of surfactants in Table 6.2 correspond to a suc-
cessive change into aggregates that correspond to a more long-range interac-
tion. If there is an upper limit for the repulsion, the solvation series is
terminated at that stage. Hence, the maximum solvated aggregate formed at a
surplus of water is a measure of the ability of the emulsifier to generate re-
pulsive interactions.
The area of the molecule is a measure of the interaction when water is
available, and may be generalized as the hydrophilicity of the molecule. The
spatial requirement of the hydrophobic part of the molecule is of course a mea-
sure of the hydrophobicity of the molecule. Consequently, there is a close link
with the classical view of emulsifiers as molecules with a balance between the
hydrophobic and the hydrophilic properties, as they are expressed in the HLB
numbers, proposed by Griffin (1949, 1979).
152 Food Emulsifiers and Their Applications
Packing constraint t
(A 0 = 23A 2 for a
Area/volume saturated hydrocarbon tail)
l l · Vhydrophoh _ A
- 0
T rhydrophob
t The packing constraint is here defined as the necessary cross section of an amphiphilic molecule
in the aggregate at the oil/water interface. This definition is Aofpacking parameter according to
Israelachvili et al. (1992, 1976, 1977).
Physicochemical Aspects of an Emulsifier Functionality 153
p- Xylene
1 2 3 4 5
Water Nonylphenoi-E09
talline phase coating the droplet surface reduces the van der Waal's attraction
and that this was an important contribution to the observed effects in the emul-
sification experiments (Friberg, 1971). However, this explanation is not a use-
ful general explanation since the emulsifier concentration in optimized food
emulsions rarely is high enough to allow for multilayer adsorption (Walstra,
1988; Dickinson, 1986). Obviously, this observation is contradictive to the
need for a separate phase of liquid-crystalline material around the droplet.
However, a correlation between the presence of, or the possibility to form, liq-
uid-crystalline phases and emulsion stability is still experimentally observed
in several systems. To stabilize an dispersion, the emulsifier should:
Stabilizing property
aggregates Micelles Bilayers Reversed
Water-continuous emulsions
Repulsive interactions Optimal Intermediate Weak
Interfacial viscosity Weak Optimal Weak
Anchoring Too water-soluble Optimal Acceptable
Oil-continuous emulsions
Repulsive interactions Weak Intermediate Optimal
Interfacial viscosity Weak Optimal Weak
Anchoring Acceptable Optimal Too oil-soluble
6.5.2 Lecithins
Lecithin is one of the most commonly used food emulsifiers, and its popularity
can be expected to grow even further due to its natural origin. Technical
lecithins, usually soybean lecithin, are always natural mixtures of various
phospholipids. The most frequent one is phosphatidylcholine (PC). The second
'"_§ .. •
4
0..
• • Lamellar phase 60 C
s 3
c Gel phase 23 C
.5 c • •
";:;'" 2 ccccc
c£
00 c
"'"0..
c c
·:n"> 0 •
:;
0.. •
~ " -1
0 5 10 15
dw(A)
Figure 6.3 The hydration repulsion between bilayers of monopalmitin in the liquid-
crystalline and gel states. (Redrawn from Pezron et al., 1991.)
156 Food Emulsifiers and Their Applications
Tetraglycerolesters:
Tetraglycerol C12 Lamellar < 20°C 55% Krog, 1990t
monolaurin Fluid isotropic 40°C
Sodium steraoyllactylate:
pHS C18 Reversed hexagonal at 45°C 40% Krog, 1990
pH7 C18 Lamellar at 42°C 60% Krog, 1990
Sorbitan esters:
Polyoxyethylene (20) C18:1 Hexagonal phase (up to 30°C)
sorbitan monooleate aud micellar solution Hall, Pethica, 1967
Polyoxyethylene (20) C18 Hexagonal phase (30 to 50°C)
sorbitan monostearate and micellar solution Hall, Pethica, 1967
above 30°C
Sorbitan stearate C18 Lamellar above 50°C - Hall, Pethica, 1967
properties of the most common phosphatides and then discuss the properties of
various mixtures.
6.5.3 Phosphatidylcholine
The phase diagram of a typical unsaturated phosphatidylcholine is displayed
in Figure 6.4. The phase diagram is characterized by a large swelling lamellar
phase. Saturated phosphatidylcholines have a phase transition temperature up
to about 40°C, whereas the corresponding temperature for unsaturated
lecithins is well below 0°C. The phase diagram of soybean PC is described in
Bergenstahl and Fontell (1983) and is rather similar to the phase diagram of
dioleoyl PC.
6.5.4 Phosphatidylethanolamine
Phosphatidylethanolamine is less hydrophilic than PC. The saturated
ethanolamines form lamellar phases that swell less than the corresponding PC
species. The phase transition temperature is about lO to 40°C above the corre-
sponding temperature of the phosphatidylcholine (Figure 6.5). The more lim-
ited ability to create long-range repulsive interactions, and thereby to defend a
large molecular area, is displayed in the tendency to form reversed hexagonal
phase with unsaturated PE species, as shown in Table 6.5.
200
Reversed
Hexagonal
~ 150
l
8 100
~ Lamellar Cubic
phase
50
Crystalline
phase
0
Water Dioleoylphosphatidylcholine
90
[J
70
G [J
•
~
c. 50 •
E 30
[J
•
~ •
10
-10
•
10 12 14 16 18 20 22
Chain length
Figure 6.5 The main transition temperature for phosphatidylcholine (PC) (•) and
phosphatidylethanolamine (PE) (D) as a function of chain length. The sources of data
are given in Table 6.4.
6.5.5 Phosphatidylinositol
The phase diagram of soybean PI and water has been determined by the author
(1991) and by Soderberg (1990). The diagram is characterized by a large
lamellar phase with an unlimited swelling. The liquid-crystalline phase is
formed below room temperature.
6.5.7 Lysophosphatides
The phase diagrams of a series of different lysophosphatides has been investi-
gated by Arvidsson et al. (1985). Lysophosphatidylcholine has the same hy-
drophilic polar group as the ordinary PC but only one of the two fatty acids.
This reduces the volume demand of the aggregate, and the packing constraint
allows for the formation of micelles and hexagonal phases.
Phosphatidyletanoleamine:
Dipalmitoyl CI6 Lamellar phase at 68°C 20% Caffrey, I9S5
Reversed hexagonal at S4°C
Dioleoyl C1S:l Lamellar below 0°C 20% Gawrish et al, 1992
Reversed hexagonal at 5°C
Soybean PE CIS l-2+ Reversed hexagonal 30% Bergenstahl, 1991
above 0°C
Phosphatidylinositol:
Soybean PI C1S:I-2+ Lamellar below 0°C Unlimited Bergenstahl, 1991
Soderberg, 1990
Phosphatidic acid:
Dioleoyl ClS:l Lamellar below 0°C Unlimited Lindblom et al., I99I
Lyso PC:
Palmi toy! CI6 Micellar solution below 0°C Unlimited Eriksson et al., I9S7
iThe data are extracted from a review of several original sources.
+Mainly.
terrnine the type of liquid-crystalline phase that develops when different phos-
phatides are allowed to interact together with water. Figure 6.6 shows the
phase diagram of dioleoyl PC and dioleoyl PE in 40% water (Eriksson et al.,
1985). The figure shows that a lamellar phase is formed when the system con-
tains mainly PC, but that about 60% PE nonlamellar phases start to form. This
change is enhanced at high temperatures. Between the hexagonal phase and
the lamellar phase is an area in which a cubic phase appears (above 50°C).
160 Food Emulsifiers and Their Applications
100
Cubic phase
80
Reversed
hexagonal
phase
G
~
60
e
::l
~
'"<l.l Lamellar phase
a
0.,
40
~
20
0
100% 50150 100%
DOPE DOPC
Composition
Figure 6.6 The phase diagram of dioleoyl PC and dioleoyl PE with 40% water.
(Redrawn after Eriksson et al., 1985.)
Water PC Water
PI
Water
Figure 6.7 The phase diagram of soybean PC, soybean PE, and water; of soybean PC,
soybean PI, and water; and of soybean PI, soybean PE, and water. (Redrawn after
Bergenstahl, 1991.) The cubic phase was not included in the original drawing, but it is
a possible interpretation of the x-ray peaks included in the paper. It is also supported
by the data from the study by Eriksson eta!. (1985).
sion, and a water-soluble emulsifier will tum the emulsion into a water-contin-
uous one. This is true for low molecular emulsifiers with a high solubility (usu-
ally in micellar aggregates), but it is also valid for polymers. However, most
likely, the concept can also, to some extent, be expanded to include emulsifiers
with just a dispersibility in either one of the phases (for instance lecithin).
Experience in this direction is exemplified in Table 6.6. However, the Bancroft
rule provide us just with the first very general directions. To proceed further we
need possibilities to rank emulsifiers quantitatively.
about 20 to 30°C above the final storage temperature [emulsification by the PIT
method (Shinoda and Saito, 1968)].
Shinoda and coworkers (Shinoda and Saito, 1968; Shinoda and Kunieda,
1983; Kunieda and Ishikawa, 1985, reviewed in Shinoda and Friberg, 1986)
have worked according to this concept and characterized a number of different
ethoxylated emulsifiers in combination with various solvents. They then found
that the PIT depends not only on the number of ethoxy groups but also on the oil
phase, indicating the importance of the solubility properties for the stability.
Emulsification experiments performed with a range of different oil-to-water
ratios show that the emulsion type is determined mainly by the emulsifier
properties and is for many systems (pure solvents!) very insensitive to the
phase ratio (Shinoda and Friberg, 1986).
It is obvious that this says a lot about the properties of ethoxylated surfactants
but its applicability to food emulsions is very limited for two main reasons:
> 13 >2
..,.,--.
~>t· Clear solution
Micelles
Lamellar phase
Reversed aggregates
This result shows that the ability to form liquid-crystalline phases corre-
sponds to the traditional HLB characterization of the emulsifiers.
Static Dynamic
The principal role of the intenacial tension is obvious. The presence of emul-
sifiers lowers the intenacial tension from about 30 mN/m for a triglyceride/water
system to between 1 and lO mN/m. Nonionic emulsifiers close to the PIT create
densely packed intenaces with very low intenacial tensions. However, the ef-
fects of the intenacial tension itself are not very large. Walstra (1983) has shown
that the droplet size is only weakly dependent on the intenacial tension.
During the homogenization, new intenaces are formed. The emulsifiers have
to diffuse to the intenaces to lower the intenacial tension during the events when
the droplets are formed. This process must be rapid to be successful, as rapid as
the time scale for the formation of the droplets. For geometrical reasons, the dif-
fusion from the surrounding phase of the droplet is much more rapid than the
diffusion from the internal liquid. This is one important contribution to the valid-
ity of the solution rules (Bancroft, PIT, HLB, and phase diagrams).
During the homogenization, the water-soluble substances in the oil phase
diffuse over to the water phase. These types of diffusion across the intenaces
create disturbances that contribute to the emulsification. In many systems, this
effect gives an increased efficiency if the emulsifier is added to the oil phase
before the emulsification. For dispersible emulsifiers (phospholipids) there are
also other reasons why it is more efficient to add the emulsifier to the oil phase
instead of the water phase. During the homogenization, phospholipids tend to
form stable liposomal dispersion in competition to the emulsification of the oil
phase. Westesen has indeed observed that a significant fraction of the phos-
pholipids in a commercial phospholipid emulsion for paranteral use is lost in
liposomal aggregates (Westesen and Wehler, 1992).
Emulsification involves an intensive shear. The shear by itself causes a
high frequency of recoalescence events. If the emulsification is to be success-
ful the formed droplets have to be protected. The repulsive interactions gener-
ated by the emulsifiers create a static protection.
The hydrodynamic interaction is crucial for the result of a collision due to
shear. The hydrodynamic interactions depend on the existence of an intenace
with an intenacial viscosity and elasticity. During the collision event, the in-
tenace close to the approaching droplet is depleted of emulsifiers due to the
streaming ofliquid. The sunactant-depleted zone will then have a higher inter-
facial tension than the surrounding emulsifier-covered areas of the droplets.
This leads to sunace diffusion in the direction opposite to the liquid flow and
ensures the hydrodynamic resistance. If the emulsifier is oil-soluble, emulsi-
Physicochemical Aspects of an Emulsifier Functionality 167
fier from the internal part of the droplet will diffuse to the depleted area and
thereby reduce the hydrodynamic protection of the droplet.
The discussion in this section has been very qualitative, but an important
point is that the emulsifiers contribute to the emulsification as well as to the
stabilization. The role of the emulsifier for the stabilization is usually difficult
to identify in the simple type of shaking experiments that are the main back-
ground to the HLB, the PIT, and the phase diagram concepts. This type of sim-
ple, and thereby effficient, experiment provides information about both the
emulsifiability and the stability with a certain emulsifier.
from a mixture of two emulsifiers, the most hydrophobic emulsifier will have the
strongest affinity to the interface. A consequence is that under competitive ad-
sorption the component with the lowest water solubility will dominate the inter-
face [e.g., the lowest critical micelle concentration (Kronberg, 1983)].
The character of the adsorbed layer, for instance its ability to generate re-
pulsive interactions, is determined by the dominating compound. The struc-
ture of the layer depends on the geometrical shape of the molecules and on
lateral interactions between the molecules in the layer. Nonionic surfactants
may form very dense layers due to head-group attraction. Ionic surfactants are
able to form extremely loose layers due to inter-head-group repulsion.
An interesting experimental observation in agreement with this relation is
that the concentration of emulsifier necessary to obtain an emulsion is much
lower for ionic emulsifiers than for nonionic emulsifiers.
In a series of emulsions, we have studied the efficiency of the emulsification
(Ostberg et al., 1995) by droplet size measurements after homogenization. The
results show that for several emulsifiers very small droplets are obtained
(about 0.2 to 0.4 Jlm). The particle size obtained depends on the concentration
of emulsifier. The nonionic emulsifiers leads to a constant particle size down to
a critical concentration below which the ability to form emulsions is strongly
reduced. The critical concentration can be compared with the thickness of the
emulsifier layer on the emulsion droplet. The apparent thickness of the emulsi-
fier layer can be estimated from the particle size and the concentration of
emulsifier (counted on the dispersed phase), if we assume that all emulsifier is
adsorbed to the interface. The apparent thickness gives the upper limit for the
adsorbed layer rather than the correct value:
volume of emulsifier
Thickness of emulsifier layer =
emulsion droplet area
Aemulsion droplet 6
ness. The properties of the ionic emulsifiers are different. These emulsifiers
are able to emulsify the emulsions down to extremely low concentrations corre-
sponding to very low surface concentrations (thin layers).
Table 6.9 The apparent emulsifier layers for various emulsifiers estimated from
equation. (From Ostberg et al., 1995.)
Estimated length
Emulsifier Radius Emulsifier-layer Curve of the emulsifier
cone (%)1 (Jlm)2 thickness (1)3 shape 4 (1)5
Dodecylbenzenesulfate
10 0.27 90 54
7 0.20 45 75
lO 0.23 59 93
1 The emulsifier concentration calculated on the oil phase.
2 The radius is shown as D(3, 2)/2.
3 The apparent emulsifier layer, estimated assuming that all emulsifier is estimated at the interface.
4 The curve shape shows the dependence for the apparent emulsifier layer of the emulsifier concen-
tration.
5 The estimated length of the emulsifier molecule is estimated from the chemical formula or from
tion. Courthaudon et al. (1991b) have shown C12E08 totally displace all ad-
sorbed ~-casein from an emulsion system. Similar effects have also been ob-
tained with emulsions formed with polysorbates (Dickinson and Tanai, 1992)
and with monoglycerides (Hall and Pethica, 1967). On the other hand, egg
yolk PC did not reduce the the adsorbed amount of ~-casein more than about
20% (Courthaudon et al., 1991a).
The adsorption of a range of plasma proteins at various phospholipid sur-
Physicochemical Aspects of an Emulsifier Functionality 171
References
Arvidsson, G., et al. (1985). Eur. ]. Biochem, 152, 753-9.
Bancroft, W.D. (1913).]. Phys. Chem., 17, SOl.
Bergenstahl, B. (1991). In Food Polymers, Gels, and Colloids (ed. E. Dickinson), Royal
Society of Chemistry London, pp. 123-131.
- - - , Claesson, P. M. (1990). In Food Emulsions (eds. K. Larsson, S. Friberg),
Marcel Dekker, New York.
- - , Fontell K. (1983). Prog. Call. Pol. Sci., 68, p. 48-52.
- - , Stenius P.J. (1987). Phys. Chem., 91, 5944-48.
Boyd, J. V., et al. (1976). In Theory and Practice of Emulsion Technology (ed. A. L.
Smith), Academic, London.
Caffrey, M. (1985). Biochemistry, 24, 4826--44.
Courthaudon, J. L., et al. (1991a).]. Agr. Food Chem., 39, 1365.
- - et al. (1991b).]. Colloid Interface Sci., 145, 390.
Darling, D., Birkett, R. J. (1987). In Food Emulsions and Foams (ed. E. Dickinson),
Royal Society of Chemistry, London.
Davies, J. T. (1957). Proc. Intern. Congr. Surf. Activity, 2d, London, 1, 426.
Dickinson, E. (1986). Food Hydrocolloids, 1, 3.
- - - , Tanai S. (1992). Food Hydrocolloids, 6, 163-71.
- - et al. (1991). Food Hydrocolloids, 4, 403-14.
Eriksson, P. 0., et al. (1987). Phys. Chem., 91, 846-63.
- - et al. (1985). Chem. Phys. Lipids, 37, 357-71.
Fontell, K. (1978). Progr. Chem. Fats Other Lipids, 16, 145-62.
- - - et al. (1968). Acta Polytechnica Scandinavica, Chapter 2, Chemistry Series III,
74, 2.
Friberg, S. (1990). In Food Emulsions (eds. K. Larsson, S. Friberg), Marcel Dekker,
New York.
- - (1971).]. Colloid Interface Sci., 37,291.
--,Mandell, L. (1970a). ]. Assoc. Off. Chem. Soc., 47, 149.
- - (1970b). ]. Pharm. Sci., 59, 1001-4.
- - , Rydhag, L. (1971). Kolloid Z. u. Polymere, 244, 233-9.
- - - , Wilton, I. (1970). Liquid crystals-the formula for emulsions, Am. Parf. and
Cosm., 85, 27-30.
- - et al. (1969). ]. Colloid Interface Sci., 29, 155-6.
Gawrish, K., et al. (1992). Biochemistry, 31, 2856-64.
Griffin, W. C. (1979). In Kirk-Othmer Encyclopedia of Chemical Technology, Wiley, New
York, Vol. 8.
- - - (1949).]. Soc. Cosmetic Chemists, 311-26
172 Food Emulsifiers and Their Applications
Hall, D.G., Pethica, D.A. (1967). In Nonionic Surfactants (ed. M.J. Schick), Marcel
Dekker, New York, p. 516.
lnoko, Y., Mitsui, T.J. (1978). Phys Soc. ]ap., 44, 1918.
lsraelachvili, J. (1992). Intermolecular and Surface Forces, Academic, London.
--eta!. (1977). Biochim. Biophys. Acta, 470, 185-201.
---eta!. (1976). ]. Chem. Soc. Faraday Transactions II, 72, 1525.
Janiak, M.J., eta!. (1979). ]. Biol. Chem., 254, 6068-78.
Krog, N. (1990). In Food Emulsion (eds. K. Larsson, S. Friberg), Marcel Dekker, New
York, p. 127.
Kronberg, B. (1983). ]. Colloid Interface Sci., 96, 55-68.
Kunieda, H., Ishikawa, N. (1985). ]. Colloid Interface Sci., 107, 122-28.
- - , Shinoda, K. (1985). ]. Colloid Interface Sci., 107, 107-21.
Larsson, K., Krog, N. (1973). Chem. Phys. Lipids, 10, 177.
Lindblom, G., et al. (1991). Biochemistry, 30, 10938-48.
Malmsten, M. (1995). ]. Colloid Interface Sci., 172, 106--15.
Ostberg, G., et a!. (1995). Colloid Surfaces A, Physichemical Engineering Asp., 94,
161-71.
Pezron, 1., eta!. (1991).]. Colloid Interface Sci., 144, 449-57.
Rydhag, L. (1979). Fette Seifen Anstrichm., 81, 168-73.
--,Wilton, I. (1981). ]. Assoc. Off. Chem. Soc., 58, 830-7.
Shinoda, K., Friberg, S. (1986). Emulsions and Solubilization, Wiley, New York.
- - - , Kunieda H. (1983). In Encyclopedia of Emulsion Technology, Vol. 1 (ed. P
Becher), Marcel Dekker, New York.
---,Saito, H. (1968). ]. Colloid Interface Sci., 30, 258-63.
Small, D. M. (1986). Handbook of Lipid Research: Physical Chemistry of Lipids,
Plenum, New York.
Soderberg, I. (1990). Structural Properties of Monoglycerides, Phospholipids and Fats
in Aqueous Systems, PhD Thesis, University of Lund, Sweden.
Tanford, C. (1973). In The Hydrophobic Effect, Wiley, New York.
Walstra, P. (1983). In Encyclopedia of Emulsion Technology, Vol. 1 (ed. P. Becher),
Marcel Dekker, New York, p. 57.
- - - (1988). In Gums and Stabilizers for the Food Industry, Vol. 4 (eds. G.O. Phillips
eta!.), IRL Press, Oxford, pp. 233-336.
Westesen, K., Wehler, T. (1992). ]. Pharm. Sci., 81, 777.
Wilton, 1., Friberg, S. (1971). Influence of temperature-induced phase transition in fat
emulsions,]. Assoc. Off. Chem. Soc., 48, 771-4.
SEVEN
7.1 Introduction
Bovine milk has been an important source of food for human beings for thou-
sands of years. Not only is milk a very nutritious food in its own right, but it is
also a very versatile starting point for many other dairy products.
Milk is a complex food emulsion and colloidal sol. Table 7.1 gives the compo-
sition of whole cow's milk. The emulsion component is composed of fat droplets
dispersed in an aqueous phase containing protein. The protein is in the form of
both casein micelles, which are themselves colloidal particles, and free in solu-
tion as whey protein. A considerable reserve of knowledge has been assembled
on the structure and properties of the milk proteins (Swaisgood, 1992). The fat
droplets are stabilized by an adsorbed layer of protein and phospholipid called
the "milk fat globule membrane" (MFGM), which is distinct from the aqueous
phase protein (Walstra and Jenness, 1984). The average composition of the
MFGM has been estimated to be about 48% protein, 33% phospholipid, and
ll% water, with the remainder made up of other minor lipid components
(Walstra and Jenness, 1984). The phospholipid fraction of the membrane is com-
posed of lecithin, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl
inositide, plasmalogens, and sphingomyelin. Phospholipids are important food
173
174 Food Emulsifiers and Their Applications
Table 7.1 Approximate composition of bovine milk. (From Walstra & Jenness, 1984,
Dairy Chemistry and Physics. Copyright © 1984, Wiley-lnterscience. Reprinted by per-
mission of John Wiley & Sons, Inc.)
emulsifiers in their own right. The contribution that they make to the stability of
the milk fat globule is not well understood, but their use as food-grade emulsi-
fiers has been the subject of extensive fundamental research (Courthaudon et al.,
1991; Dickinson et al., 1993a; Dickinson and Iveson 1993).
Until this century the conversion of milk to butter, cream, ice cream, and
various types of cheese had been more of a craft than a science. It is only rela-
tively recently that an albeit incomplete understanding of these processes has
been available. It is now understood that the formation of these milk-based
products is a consequence of either the destabilization of the dispersed-phase
fat droplets (as in butter, ice cream, and whipped cream) or of the dispersed
aqueous-phase proteins (as in cheese). Emulsifiers play an important role in
the formation of structure in many dairy products.
In the following pages, the major emulsifier-containing dairy and imitation
dairy products will be reviewed. A brief description of their production will be
given where relevant, with emphasis on the role that emulsifiers play in the
formation and stability of the product.
and Barfod, 1990; Barfod et al., 1991). Before describing the role of emulsi-
fiers in ice cream structure formation, it is pertinent to consider the structure
of the adsorbed layer formed around the fat droplet in the ice cream premix
emulsion. During the homogenization stage the ice cream mix is subject to
high shear. This results in the disruption of the fat phase into small droplets.
Surface-active components of the mix will adsorb onto the nascent-oiVwater
interface, lowering the interfacial tension and thus stabilizing the emulsion
droplets. The exact composition of the interfacial layer will depend on the pro-
portions of each type of surface-active component and their relative surface
activities. Recent research has thrown light on the competitive adsorption be-
tween different milk proteins (Dickinson, 1986; Dickinson et al., 1988b;
Dickinson et al., 1989a; Dickinson et al., 1989c; Euston, 1989; Dickinson et
al, 1990b; Dickinson, 1992), milk proteins and emulsifiers (Dickinson and
Woskett, 1988a; Dickinson et al., 1990a; Dickinson, 1992; Euston et al.,
l995a; Euston et al., 1995b), and on the cooperative adsorption between pro-
teins and polysaccharide stabilizers (Dickinson and Euston, 1990; Dickinson,
1992; Dickinson, 1993). These studies have identified that, in general, the
more surface-active component will predominate at the fat-droplet interface.
Thus the more surface-active protein in a mixture will dominate the adsorbed
layer initially, low molecular weight emulsifiers will, generally, displace pro-
tein from the surface with time, and under certain conditions polysaccharides
can interact with proteins and/or emulsifiers to contribute to the structure of
the adsorbed layer (Bergenstahl et al., 1992). The adsorbed layer of the ice
cream emulsion will be a composite of these functional ingredients.
The key to the formation of ice cream structure is the formation of a stable
foamed product. In ice cream this is achieved in two ways. The foam in ice
cream is not a typical protein-stabilized foam, where air bubble stabilization is
achieved by protein adsorption at the air/water interface. The initial stabiliza-
tion of the foam network may indeed proceed by this mechanism, but prior to
freezing, the foam structure is stabilized primarily by the partial coalescence
of emulsion fat droplets at the air bubble interface. Figure 7.1 is a cryo-SEM
micrograph of ice cream. Adsorption of fat globules at the air bubble surface is
clearly visible. For this to be able to occur, the emulsion must be relatively un-
stable to the shearing forces exerted on the mix during air incorporation.
Several workers have attempted to demonstrate the link between emulsion co-
alescence stability and the mechanical or viscoelastic properties of the ad-
sorbed protein layer (Doxastakis and Sherman, 1984; Rivas and Sherman,
Emulsifiers in Dairy Products and Dairy Substitutes 177
1984; Dickinson and Stainsby, 1988; Dickinson et al., 1988a). Dickinson et al.
(1988a) have shown a correlation between the surface shear viscosity of ad-
sorbed layers of various proteins at the planar oil/water interface, and the coa-
lescence stability of emulsions made with these proteins. The higher the
interfacial viscosity, the more stable the resultant emulsion under perikinetic
conditions. It has also been demonstrated that the orthokinetic stability of pro-
tein-stabilized emulsions (stability under turbulent or shearing conditions) is
reduced by the presence of low molecular weight emulsifiers (Chen et al.,
1993; Dickinson et al., 1993; Dickinson and Williams, 1994). The explanation
for this lies in the ability of the low molecular weight emulsifier to displace
protein from the fat-droplet smface, thus reducing the mechanical strength of
the adsorbed layer. Emulsifiers present at concentrations too low to cause sig-
nificant protein displacement can interfere with interprotein interactions
within the adsorbed layer and reduce the interfacial viscosity in this way
(Dickinson et al., 1990a).
Figure 7.1 Cryo scanning electron micrograph of ice cream. Note the partial coverage
of the air bubble (AB) surface by fat globule (FG) and the partially coalesced fat glob-
ules (PC) in the foam lamellae. (Courtesy of A.B. McKenna.)
178 Food Emulsifiers and Their Applications
30
Q)
rJ)
cc
-
..r::
c.
.....cc 20
c
-
c
Q)
0....
c.
eft-
Q)
> 10
·;:;
cc
Q)
a:
0 2 3 4 5 6 24
Figure 7.2 Changes in amount of protein adsorbed to fat globules in an ice cream mix
during the aging period at 5°C. The amount of protein bound in the fat phase is calcu-
lated relative to the total protein in the mix. (From Krog, N., Barfod, N.M., AIChE
Symposium Series, 86, 1-6. Reproduced with permission of the American Institute of
Chemical Engineers.© 1990 A!ChE. All rights reserved.)
believe that in ice cream insufficient emulsifier is present in the system for liq-
uid-crystalline phases to form. For this to occur the emulsifier must be ad-
sorbed on the fat droplet surface at a concentration far greater than that
required for monolayer surface coverage. This is not the case in ice cream
emulsions (Darling and Birkett, 1987).
Once the emulsion has been destabilized in the freezer, partial coales-
cence of the fat droplets has to occur at the air/water interface to partially
stabilize the foam structure prior to freezing. Research (Boode, 1992; Boode
and Walstra, 1993) has indicated that the crystal structure of the fat in emul-
sion droplets is important in determining their susceptibility to partial coa-
lescence. Van Boekel (1980) has shown that when fat crystals form at the
surface of emulsion droplets, and are large enough to penetrate the adsorbed
layer, a lipid bridge can form between two droplets that are in contact with
each other. The proportion of solid fat in emulsion droplets is important in
determining instability (Walstra, 1987). If the majority of the fat is solid, coa-
lescence, or partial coalescence will not occur, and the droplets will be sta-
ble. Similarly, if the droplets contain a very low proportion of solid fat,
coalescence can occur. If the solid fat content is in the approximate range 10
to 50%, partial coalescence is possible. A proportion of the emulsion droplet
is required to be in the form of liquid fat for partial coalescence to occur.
Walstra (1987) envisages a partial coagulum offat droplets as being held to-
gether by necks of liquid oil.
When fat crystallizes in dispersed emulsion droplets, considerable super-
cooling can be observed. Crystallization of fats occurs at nucleation points that
already exist in the fat phase. These nucleation points occur relatively infre-
quently in emulsion droplets where the fat is dispersed into a large number of
small droplets. Consequently, in dispersed systems the triglyceride needs to be
cooled below its bulk phase freezing point before crystallization is initiated.
Emulsifiers in adsorbed monolayers can act as templates for the surface crys-
tallization of triglycerides. Emulsifiers containing saturated hydrocarbon
chains have been shown to be good initiators of fat crystallization, whereas
those with unsaturated hydrocarbon chains are not as good (Berger, 1990;
Barfod et al., 1991). Figure 7.3 gives the solid fat content (SFC) as a function
of time for model ice cream emulsions stored at 5°C. Both saturated (GMS) and
unsaturated (GMO) emulsifiers initiate crystallization compared to control
emulsions with no emulsifier, but the SFC of GMS-containing emulsions is al-
ways greater than for those containing GMO.
Emulsifiers in Dairy Products and Dairy Substitutes 181
100
.....
c:
Q)
..... 80
c:
0
u
..... 60
.....co
"0
0Cll 40
~
20
0 2 3 4 24
Hours at 5°C
Figure 7.3 Recrystallization of fat phase of ice cream emulsions without emulsifier
(control) or with saturated (GMS) or unsaturated (GMO) monoglycerides after cooling to
5°C and aging. (From Barfod et al., 1991, and reproduced with permission.)
Darling and Birkett (1987) point out that in a mixed triglyceride system,
such as is found in milk fat, single discrete crystals are unlikely to form under
the rapid cooling conditions used in ice cream manufacture. They have shown
that in a cooled vegetable oil emulsion, concentric layers of triglyceride crys-
tals are formed at the surface of oil droplets. These contain imperfections that
may be due to dislocations or recrystallization processes, and these are likely
to cause the destabilizing effect.
The ability of triglyceride crystals to penetrate the adsorbed layer depends
on a number of factors. The surface tension between crystal and oil, crystal
and water, and oil and water will determine how far into the oil phase the crys-
tal will penetrate (i.e., if it is preferentially wetted by the oil or water phase).
Emulsifiers, by way of their surface activity, will alter these surface tensions,
and this may result in the crystals being able to penetrate further into the
aqueous phase. This would lead to a decrease in emulsion stability. The des-
orption of protein by emulsifier also aids destabilization by reducing the thick-
ness of the layer through which fat crystals have to penetrate. The polymorphic
form of the fat crystal will also play a role in fat-droplet instability.
Triglycerides can exist in three general polymorphic forms, the a, ~. and W
polymorphs. When cooled from a melt, triglycerides will generally form a-type
182 Food Emulsifiers and Their Applications
crystals. These are not stable (Larrson and Dejmek, 1990) and will transform
into a Wpolymorph and subsequently to the stable ~ polymorph. Having been
formed at lower temperature, a crystals contain triglyceride in a more disor-
dered liquid-like arrangement. These disordered crystals are softer and are
able to deform and follow the contours of the fat droplet more easily.
Consequently, they are less likely to penetrate the adsorbed layer. The ~-crys
talline structure is more solid-like, with the triglyceride molecules arranged in
ordered arrays. The ~ crystals have a greater mechanical strength and are un-
able to deform to the shape of the fat droplets. This leads to their bursting out
of the droplet into the aqueous phase (Darling, 1982).
In practice, two types of emulsifier are commonly used in ice cream: mono-
and diglycerides and polyoxyethylene derivatives of glycol or glycol esters, for
example polysorbates (Keeney, 1982). Sucrose esters have also been evaluated
and have been found to be suitable as ice cream emulsifiers (Buck et al.,
1986). Mono- and diglycerides and polysorbates are usually all found in cur-
rent ice cream emulsifier blends. The explanation for this lies in the relative
abilities of polysorbates and mono- and diglycerides as emulsion destabilizers,
or as foam-forming agents. Polysorbates are far more efficient at displacing
protein from the oiVwater interface than are mono- and diglycerides and thus
are better emulsion destabilizers (Keeney, 1982). Mono- and diglycerides are
better foaming agents and thus are able to aid the formation of the initial foam
prior to fat-droplet agglomeration at the air/water interface (Keeney, 1982). A
second factor is the differing abilities of emulsifiers to influence fat crystalliza-
tion. Figures 7.2 and 7.3 show that whereas GMO is able to displace more pro-
tein from the fat globule surface during aging than does GMS, GMS initiates
more fat crystallization than GMO. Use of a mixed emulsifier system would
also allow optimum protein displacement combined with optimum fat crystal-
lization.
In summary, nonprotein emulsifiers are important in ice cream in several
respects:
l. They promote protein desorption from the surface of fat droplets, both by
their higher relative surface activity and the possible formation of liq-
uid-crystalline mesophases.
2. They can act as nucleation points for surface crystallization of triglyc-
erides.
3. They may promote fat crystal penetration of the adsorbed protein layer by
alteration of the surface tension between various phases.
Emulsifiers in Dairy Products and Dairy Substitutes 183
4. They may help in the initial formation and stabilization of the ice cream
foam prior to partial fat-globule coalescence and freezing. Monoglycerides
are particularly good at this function.
Figure 7.4 Cryo scanning electron micrograph of whipped cream. Note the greater cov-
erage of the air bubble (AB) surface by adsorbed fat globules (FG) than occurs in ice
cream (Figure 7.1). (Courtesy ofA.B. McKenna.)
ciallayer. The adsorbed layer in homogenized cream has been found to consist
mainly of caseins, with smaller amounts of ~-lactoglobulin and ~-lactalbumin
(Anderson et al., 1977; Darling and Butcher, 1978; McPherson et al., 1984).
This is consistent with the available data on competitive adsorption between
casein and whey proteins in model systems (Euston, 1989; Dickinson et al.,
1990b). In the early stages of whipping, before fat-globule adsorption and par-
tial coalescence occurs to any great extent, the air bubbles are stabilized by
adsorbed milk serum proteins. Since this air/water interface is composed of the
same proteins that surround the fat/water interface in homogenized cream, the
difference in interfacial tension between the two interfaces is not very great
(Anderson and Brooker, 1988). Interfacial tension differences have already
been put forward as a driving force for fat-globule adsorption at the air/water
interface in ice cream (Section 7.2). Because of the small interfacial tension
differences between fat globule and air bubble in homogenized cream, the dri-
ving force for fat-globule adsorption is low. In nonhomogenized milk the inter-
facial tension differences between a fat globule stabilized by MFGM and an air
bubble stabilized by cream serum proteins are sufficient to act as the driving
force for fat-globule adsorption at the surface of air bubbles. Of course, this is
also aided by the presence of fat crystals at the fat-globule surface and by the
shearing forces introduced during whipping.
The importance of the fat phase manifests itself in two ways. As in ice
cream (Section 7.2), fat crystals are known to be important in the shear-in-
duced coalescence of the fat globules (Darling, 1982), and the presence of a
certain amount of liquid fat is a prerequisite for good whipping properties.
Bucheim (1986) put forward the idea that the interfacial layer surrounding the
fat droplets ruptures when they collide during agitation. The subsequent
spreading of liquid fat is the first stage in destabilization by aggregation of ad-
jacent droplets. The importance of the solid fat content of the fat globules has
been demonstrated by Darling (1982), who observes a direct correlation be-
tween the SFC and whipping time in natural cream.
Recombining technologies are becoming an increasingly important process
for making whipping cream bases. These would encounter the same problems
as homogenized cream (i.e., the similarity in composition and interfacial ten-
sion between air/water and oil/water interfaces), if they were formulated with
milk protein as the only surface-active material. For this reason, recombined
whipping creams, and indeed homogenized natural creams contain added low
molecular weight emulsifiers. These will alter the composition of the fat-
186 Food Emulsifiers and Their Applications
l. They destabilize the cream through their ability to displace protein from
the oil/water interface. This changes the adsorbed layer composition
and interfacial tension of the fat droplet.
2. They may destabilize the emulsion through their ability to form lyotropic
liquid-crystalline mesophases and the subsequent phase transforma-
tions that occur to form stable crystalline forms.
3. They may participate in the initial foam stabilization.
4. They aid in the formation of fat crystals at the fat-droplet surface, which
crystals are essential for fat-globule partial coalescence.
Figure 7.5 Cryo transmission electron micrograph of whipped cream containing Tween
80 as emulsifier. A partial agglomerate of fat globules is shown. The spikes emanating
from the fat globule (FG) are fat crystals (FC) and can clearly be seen to bridge between
two fat droplets (B). These spikes are, apparently, features of Tween-containing sys-
tems. (Courtesy of A.B. McKenna.)
tics and foam structure. The emulsion becomes unstable due to spontaneous
recrystallization of the supercooled fat. The destabilization of the emulsion is
probably promoted by the temperature-dependent desorption of protein from
the surface, followed by coalescence. This makes crystallization of the super-
cooled fat more likely, due to the increased probability of nucleation sites
(Bucheim et al., 198S).
Scanning electron microscopy studies (Bucheim et al., 198S) show that the
final structure of the aerated whipped topping is stabilized by a layer of crys-
talline fat of about 0.1jlm thickness. The aqueous phase lamellae between air
bubbles also contains large proportions of crystalline fat, with smaller propor-
tions of relatively intact fat globules.
The kinetics of fat crystallization and emulsion destabilization depend on
the type of emulsifier used in the formulation. Bucheim et al. (198S) have in-
vestigated the effect of distilled propylene glycol monostearate (PGMS), dis-
tilled unsaturated monoglycerides (glycerol monooleate, GMO), and distilled
saturated monoglycerides (glycerol monostearate, GMS) on the structure of
whipped toppings. Only PGMS is typically used in commercial formulations.
Figure 7.6 shows that all the emulsifiers promote fat crystallization when com-
pared to toppings without added emulsifier. The effect of PGMS and GMO,
however, was greater than for GMS. The increased emulsion destabilization
caused by the enhanced fat recrystallization led to PGMS- and GMO-contain-
ing whipped toppings being more stable than those made from GMS-contain-
ing emulsions (Bucheim et al., 198S). The GMS-containing reconstituted
topping emulsion is too stable to allow consequent stabilization of incorporated
air (whipping).
The importance of protein desorption on the whipped topping structure and
stability has been demonstrated by Krog and coworkers (Krog et al. 1986;
Barfod and Krog, 1987). Table 7.4 gives the percentage protein contents of the
fat and aqueous phases of whipped topping powders reconstituted at soc and
30°C, and containing PGMS, GMS, or no added emulsifier. At soc in the ab-
sence of emulsifier, almost one-quarter of the protein is associated with the fat
phase. This falls to 1.3% when PGMS is present and 7.7% with GMS added.
The temperature dependence of protein displacement is also evident. At 30°C,
over 40% of the protein is in the fat phase when emulsifier is absent, but this
drops to about 33.7% in the presence of PGMS and 12.3% when GMS is in-
cluded in the formulation (Krog et al., 1986). The GMS-containing reconsti-
tuted toppings have a significant proportion of protein still associated with the
190 Food Emulsifiers and Their Applications
40
~
.....
c
Q)
.....
c
0 30
u
.....Ctl
......
"0
0
(/)
20
... ...
...
10
0 10 20 30
Figure 7.6 Crystallization of supercooled lipid fractions in topping emulsions with dif-
ferent surfactants, reconstituted (I :3) in deuterated water (D20), measured by pulsed-
NMR at l5°C. e GMO (Dimodan 0), • PGMS (Promodan SP), o GMS (Dimodan PV),
A no surfactant added. [From Bucheim et al. (1985). Reprinted by permission of
Scanning Microscopy International.]
Table 7.4 Distribution of protein between the fat cream phase and the water
phase of centrifuged topping emulsion. (From Barfod and Krog, 1987. Reprinted by
permission of the American Oil Chemists Society.)
fat-droplet surface, even after aging of the emulsions at 5°C. This is obviously
enough to form an adsorbed layer strong enough, in combination with the lower
degree of fat recrystallization, to prevent stabilization of the incorporated air
bubbles. This is in contrast to the situation in ice cream and whipping cream,
where GMS is capable of destabilizing the fat emulsions to a degree that par-
tial coalescence can occur. Darling and Birkett (1987) point out that the level
Emulsifiers in Dairy Products and Dairy Substitutes 191
stage process and the two-stage process. Figure 7. 7 presents flowcharts for
both processes. The main difference between the two processes lies in the
stage at which the alcohol is added. In the single-stage process this is prior to
homogenization, whereas in the two-stage process it is after homogenization.
Banks and Muir (1988) found that homogenization in the presence of alcohol
leads to the formation of fewer large fat globules, and as such is preferable in
terms of emulsion stability. A characteristic of cream liqueur production is the
harsh homogenization conditions used (two passes at 300 bar). This results in
a product in which more than 97% of the fat droplets have a diameter less than
0.8 f..Lm. A second factor favoring formation of smaller fat droplets is the signif-
icant lowering of interfacial tension observed at the oil/water interface when
alcohol is added to the aqueous phase (Bullin et al., 1988; Dickinson and
Woskett, 1988; Burgaud and Dickinson, 1990). As a result of the very fine
droplet size, the protein in the added cream has to be supplemented by sodium
caseinate (to a fat-to-caseinate ratio of approximately 5:1) to provide adequate
coverage of the newly formed fat surface by protein (Banks et al., 1981). The
fine particle size of the dispersed fat droplets gives the product an excellent
stability with respect to creaming. Banks et al. (1981) have noted no signs of
creaming in liqueurs with a composition within the range quoted in Table 7.5
after 12 months storage. The high level of added sodium caseinate, however,
leads to cream liqueur emulsions being unstable in acid environments. This
means that they are not suitable for combination with acidic beverage mixers
such as lemonade. A cream liqueur that is stable in an acid environment can
be made by replacing the sodium caseinate with GMS. The emulsifier replaces
milk protein as the primary emulsion stabilizer at the oil/water interface, and
the nonadsorbed protein is unable to aggregate the fat droplets when exposed
Emulsifiers in Dairy Products and Dairy Substitutes 193
Single-Stage Process
Two-Stage Process
Cream Base
Homogenize
2 x 300 bar, 55"C
coolto<21rC
Homogenized Base
Gentle mixing
Alcohol
Product
Figure 7.7 Flow diagrams for the process of manufacture of a cream liqueur in (a) a
single stage and (b) two stages. (From Banks and Muir, 1988. Reprinted by permission
of Elsevier Applied Science Publishers.)
to acidic surroundings (Banks and Muir, 1988). Acid stability in this type of
product is gained at the expense of emulsion stability and shelf life. In prac-
tice, legal limits in some countries set the concentration of GMS at no more
than 0.4 wt%, and so total replacement of caseinate by GMS is not feasible.
Many manufacturers add low concentrations of GMS as well as sodium ca-
seinate to cream liqueur formulations. Dickinson and coworkers (1989b) have
shown that, in addition to displacing some, but not all of the milk protein from
the fat droplet surface, which presumably infers some acid stability on the
product, GMS also improves the creaming stability of a model cream liqueur.
When model cream liqueurs were stored at room temperature for 12 weeks, no
creaming was observed with added GMS concentrations above 0.5 wt%. Below
this level of added GMS a reduced degree of creaming was observed compared
to control samples with no emulsifier (Dickinson et al., 1989b). The increased
creaming stability was associated with rheological changes in the emulsifier
aqueous phase. At low CMS concentrations the emulsions exhibit Newtonian
behavior, whereas above 0.5 wt%, a clear yield stress is found. Dickinson et al.
194 Food Emulsifiers and Their Applications
150 . ------------------------------,
0 2
GMS concentration (wt%)
Figure 7.8 Effect of GMS on the shelf life of simulated cream liqueurs on storage at
45°C. The time for serum separation to first become visible is plotted against the GMS
concentration. Different symbols refer to separate experiments.(Based on Dickinson et
al., 1989. Reprinted by permission of the Institute of Food Technologists.)
Emulsifiers in Dairy Products and Dairy Substitutes 195
syneresis when stored for any length of time. This leads to separation of the
aqueous phase and formation of a distinct, clear serum layer at the bottom of
the sample container. Whether this syneresis will occur at room temperature is
not certain, and Dickinson et al. (1989b) stress that a correlation between the
shelf life at 45°C and that at room temperature may not follow. Cream liqueurs
stored under ambient conditions can have shelf lives of several years. Clearly
these are likely to be consumed before serum separation becomes evident.
Since the legal limits on the amount of emulsifier that can be added are set at
about 0.4 wt%, the problem of gel syneresis is unlikely to be encountered. At a
level of 0.4 wt% added GMS, creaming under gravity would not be eliminated
completely, but would be reduced to a level acceptable to the consumer
(Dickinson et al., 1989b).
Zadow (1982) states that emulsifiers and stabilizers are required only if the
product is to be given a high heat treatment (a UHT process or steam injec-
tion). This may indicate a role for the emulsifiers in protecting the protein or
emulsified fat from heat damage. Evidence exists to support this hypothesis
and will be dealt with in more detail in Section 7.8.
Whereas coffee cream and recombined cream are used in a liquid form, cof-
fee whiteners based on vegetable fat are also popular in dry powder form. Typical
formulations for liquid and dry powder coffee whiteners are given in Table 7.7.
In the past the latter process was, generally, less popular because of prob-
lems with the oxidative stability of the fat in the powder during storage.
Advances in gas packing of powders, more regular shipping, and use of cooler
storage facilities have removed this obstacle. Zadow (1982) noted that the
choice of whether to recombine or reconstitute WMP depends on the export
strategy of a particular manufacturer. During the late 1970s, an increase in the
production of reconstituted WMP was seen. This corresponded to a change
from a butter!SMP-orientated export industry to a cheese/WMP-orientated ex-
port strategy in countries such as New Zealand and Australia (Zadow, 1982).
In the recombination process, AMF, SMP, and water are recombined to give
a product with the same fat and protein content as whole milk.
The recombination process has been described by Kieseker (1983). The skim
milk powder is dissolved in the water at 40 to 55°C. The fat is added in a molten
state, and the mixture is homogenized at 14.0 to 17.5 MPa for the first stage and
at 3.5 MPa at 55 to 60°C in the second stage. The milk is then subjected to one
of three heat treatments: pasteurization at 72.2°C for 15 seconds; UHT process-
ing at 135 to 150°C for 2 to 5 seconds; or in-can sterilization (e.g., 120°C for lO
minutes). UHT processing can be by either direct steam injection for rapid heat-
ing or indirect heating in a plate or tubular heat exchanger.
Many manufacturers add low molecular weight emulsifiers to the formulation,
particularly mono- and diglycerides (Zadow, 1982; Kieseker, 1983; Sjollema,
1987). Emulsifiers in the form of phospholipids can also be added through the
practice of replacing up to 20% of the SMP with buttermilk powder (BMP)
(Zadow, 1982; Kieseker, 1983; Sjollema, 1987) to give an improved taste.
It is claimed that emulsifiers aid in the formation of the milk fat emulsion
Emulsifiers in Dairy Products and Dairy Substitutes 201
ria and bacterial spores (Tsuchido et al., 1981; Tsuchido et al. 1983), and in-
crease the heat stability of bovine serum albumin (Makino and Moriyama,
1991 ). It appears that these functions are a result of their ability to bind to pro-
teins (Clark et al., 1992; Fontecha and Swaisgood, 1994).
7.10 Summary
Emulsifiers are very versatile food additives. They can be used as aids to
emulsion formation (e.g., in coffee whiteners/creamers and recombined prod-
ucts), or in contrast, as emulsion destabilizers as in ice cream, whipping
cream, and whipped toppings. These two functions rely on the classical ability
of emulsifiers to act as surface-active agents. In this way they can influence the
formation and stabilization of the fat-droplet adsorbed layer and the composi-
tion of this layer. This ability of emulsifiers to displace protein from the fat-
droplet surface also, probably, accounts for the increase in heat stability of
concentrated milks when phospholipids are added.
In a similar vein, displacement of adsorbed caseinate by GMS in cream
liqueurs can be used to give increased acid stability to these products. A sec-
ondary function of the GMS in cream liqueurs is its ability to interact with pro-
teins, thereby forming a weak gel in the aqueous phase. The associated increase
in viscosity gives improved creaming stability. The ability of the emulsifier SSL
to interact with the proteins in caseinate is also exploited in coffee whiteners.
The replacement of sodium caseinate in powdered coffee whitener is achieved
by using SSL. It has been hypothesized (Leo and Betscher, 1971) that this is pos-
sible because of the increased mechanical strength of a protein/SSL adsorbed
layer caused by emulsifier/protein interactions.
In processed cheese, the ability of charged emulsifiers to interact with proteins
in the cheese matrix may prove a useful way of controlling cheese texture. This
would introduce a way of reducing the concentration of emulsifying salts such as
mono- and polyphosphates. The final, but very important, function of some emul-
sifiers is their ability to act as initiators of fat crystallization. This is a particularly
important function in whipped products, and in combination with protein dis-
placement forms the basis of the formation of the whipped foam structure.
A wide range of emulsifiers allowed for food use can be added to achieve the
above effects. Of late, consumer opinion has been focused on the "unnatural"
nature of synthetic emulsifiers. There is a slow push toward the replacement of
synthetic emulsifiers with natural emulsifiers such as milk and soy phospho-
204 Food Emulsifiers and Their Applications
lipids, and milk fat-derived mono- and diglycerides. The future may see a large
increase in the use of products such as BMP, which is rich in natural milk phos-
pholipids as well as protein, and milk fat that has been enriched in mono- and
diglycerides by processes such as controlled glycerolysis of triglycerides.
Acknowledgments
I would like to thank the following people: Drs. Jeremy Hill, Peter Munro, and
Mike Boland (NZDRI) and Dr. Susan Euston (Massey University) for proof-
reading this manuscript; Dr. David Newstead (Milk Powder Technology
Section, NZDRI) for valuable discussion and comments on UHT fouling and
heat stability of concentrated milks; Dr. Siew-Kim Lee (Cheese Technology
Section, NZDRI) for comments on and suggestions for the section on processed
cheese; Mr. Bing Soo (Food Systems Section, NZDRI) for supplying informa-
tion on coffee whitener/creamer; Mr. Tony McKenna (Food Science Section,
NZDRI) for supplying the electron micrographs used in Figures 7.1, 7.4, and
7.5; Vanessa Ellery (Graphics Unit, Information Centre, NZDRI) for prepara-
tion of Figures 7.2, 7.3, 7.6, and 7.7. Finally, I would like to thank the New
Zealand Dairy Board for permission to publish this work.
References
Abrahamsson, K., et al. (1988). Effects of homogenization and heating conditions on
physico-chemical properties of coffee cream, Milchwissenschaft, 43, 762-765.
Anderson, M., Brooker, B.E. (1988). Dairy foams, in Advances in Food Emulsions and
Foams (eds. E. Dickinson and G. Stainsby). Elsevier, London, pp. 221-255.
- - - et al. (1977). Changes during storage in stability and composition of ultra-heat-
treated aseptically-packed cream of 18% fat content, Journal of Dairy Research, 44,
111-123.
Arbuckle, W.S. (1986). Ice Cream, 4th ed., AVI, Westport, CT.
Banks, W., Muir, D.D. (1988). Stability of alcohol containing emulsions, in Advances in
Food Emulsions and Foams (eds. E. Dickinson and G. Stainsby). Elsevier, London,
pp. 257-283.
- - - , Wilson, A.G. (1981). The formulation of cream-based liqueurs, Milk Industry
83, 16--18.
Barfod, N.M., Krog, N. (1987). Destabilization and fat crystallization of whippable
emulsions (toppings) studied by pulsed NMR, Journal of the American Oil Chemists
Society, 64, 112-119.
- - - et al. (1991). Effects of emulsifiers on protein-fat interaction in ice cream mix
during aging I: Quantitative analyses, Fat Science and Technology, 93, 24--29.
Barratt, M.D., Rayner, L. (1972). Lysolecithin-casein interactions. I. Nuclear magnetic
Emulsifiers in Dairy Products and Dairy Substitutes 205
resonance and spin label studies, Biochimica et Biophysica Acta, 255, 974-980.
Bergenstahl, B., et a!. (1992). Adsorption structures in emulsions, in Emulsions -A
Fundamental and Practical Approach (ed. J. Sjoblom ), NATO ASI Series C, 363,
51-60.
Berger, K.G. (1990). Ice cream, in Food Emulsions (eds. K. Larsson and S. Friberg),
Marcel Dekker, New York, pp. 367-444.
Beuchat, L.R. (1980). Comparison of anti-Vibrio activities of potassium sorbate,
sodium benzoate and glycerol and sucrose esters of fatty acids, Applied and
Environmental Microbiology, 39, 1178-1182.
Boode, K. (1992). Partial Coalescence in Oil-in-Water Emulsions, PhD Thesis,
Wageningen Agricultural University, The Netherlands.
- - - , Walstra, P. (1993). Kinetics of partial coalescence in oil-in-water emulsions, in
Food Colloids and Polymers: Stability and Mechanical Properties (eds. E. Dickinson
and P. Walstra), Royal Society of Chemistry, London, pp. 23-30.
Bucheim, W. (1986). Membranes of milk fat globules-ultrastructural, biochemical and
technological aspects, Kieler Milchwirtschaftliche Forschungsberichte, 38, 227-246.
- - - et a!. (1985). Relation between microstructure, destabilization phenomena and
rheological properties of whippable emulsions, Food Microstructure, 4, 221-232.
Buck, J.S., et a!. (1986). Evaluation of sucrose esters in ice cream, Journal of Food
Science, 51, 489-493.
Bullin, S., et a!. (1988). Stability aspects of casein-containing emulsions: effect of
added alcohol or dextran, in Gums and Stabilizers for the Food Industry (eds. G.O.
Phillips eta!.), IRL Press, Oxford, Vol. 4, pp. 337-345.
Burgaud, 1., Dickinson, E. (1990). Emulsifying effects offood macromolecules in pres-
ence of ethanol, Journal of Food Science, 55, 875-876.
Carie, M., et a!. (1985). Effects of emulsifying agents on the microstructure and other
characteristics of process cheese-a review. Food Microstructure, 4, 297-312.
Chen, J., et a!. (1993). Interfacial interactions, competitive adsorption and emulsion
stability, Food Structure, 12, 135-146.
Clark, D. C., eta!. (1992). The interaction of sucrose esters with ~-lactoglobulin and~
casein from bovine milk, Food Hydrocolloids, 6, 173-186.
Conley, A.J., Kabara, J.J. (1973). Antimicrobial action of esters of polyhydric alcohols,
Antimicrobial Agents Chemotherapy, 4, 501-506.
Courthaudon, J.-L., et a!. (1991). Competitive adsorption of lecithin and ~-casein in
oil-in-water emulsions, Journal of Agriculture and Food Chemistry, 39, 1365-1368.
Darling, D.F. (1982). Recent advances in the destabilization of dairy emulsions,
Journal of Dairy Research, 49, 695-712.
---,Birkett, R.J. (1987). Food colloids in practice, in Food Emulsions and Foams,
(ed. E. Dickinson), Royal Society of Chemistry, London, pp.l-29.
- - - , Butcher, D.W. (1978). Milk fat globule membrane in homogenized cream,
Journal of Dairy Research, 45, 197-208.
Dickinson, E. (1986). Mixed proteinaceous emulsifiers: review of competitive protein
adsorption and the relationship to food colloid stabilization, Food Hydrocolloids, 1,
3-23.
- - - (1992). Structure and composition of adsorbed protein layers and the relation-
ship to emulsion stability, Journal of the Chemical Society Faraday Transactions,
889. 2973-2983.
206 Food Emulsifiers and Their Applications
Si, J.Q. (1991). The production of dairy analogue products using emulsifiers, stabilizers
and flavours, in Milk Fat Production. Technology and Utilization (eds. K.K. Rajah
and K.J. Burgess), The Society of Dairy Technology, Huntingdon, pp. 112-121.
Sims, R.J. (1989). Spray dried emulsions, in Food Emulsifiers, Chemistry, Technology,
Functional Properties and Applications (eds. G. Charalambous and G. Doxastakis),
Elsevier, Amsterdam, pp. 495-509.
Singh, H., Creamer, L.K. (1992). Heat stability of milk, in Advanced Dairy Chemistry
Volume 1. Proteins (ed. P.F. Fox), Elsevier, London, pp. 621--656.
- - - , Tokley, R.P. (1990). Effects of preheat treatments and buttermilk addition on
the seasonal variations in the heat stability of recombined evaporated milk and re-
constituted concentrated milk, The Australian Journal of Dairy Technology, 45,
10-16.
---eta!. (1992). Influence of incorporation of soya lecithin into skim milk powder
on the heat stability of recombined evaporated milk, The Australian Journal of
Dairy Technology, 47, 33-37.
Sjollema, A. (1987). Recombination of milk and dairy ingredients into milk, cream,
condensed milk and evaporated milk, in Milk-The Vital Force, Reidel Publishing,
Boston, MA, pp. 251-257.
Swaisgood, H. E. (1992). Chemistry of the caseins, in Advanced Dairy Chemistry 1:
Proteins (ed. P.F. Fox), Elsevier, London, pp. 63-110.
Thomas, M.A., et a!. (1980). Effect of emulsifying salts on objective and subjective
properties of processed cheese, Journal of Food Science, 45, 458-459, 466.
Thome, K E., Eriksson, G. (1973). The foaming properties of cream 2. The effect of sur-
face active agents, especially phospholipids and lipoproteins, on the whippability of
cream, Milchwissenschaft, 28, 554-558.
Tsuchido, T., et a!. (1987). Lysis of Bacillus subtilis cells by glycerol and sucrose esters
of fatty acids, Applied and Environmental Microbiology, 53, 505-508
---eta!. (1981). Death kinetics of Escherichia coli in a combined treatment with
heat and monolaurin, Journal of Food Safety, 3, 57--68.
- - - et a!. (1983). Inhibitory effect of sucrose esters of fatty acids on intact and
heated bacterial spores, Journal of Antibacterial and Antifungal Agents, 11,
567-573.
Van Boeke!, M.A.J.S. (1980). The Influence of Fat Crystals in the Oil Phase on Stability
of Oil-in-Water Emulsions, PhD Thesis, Wageningen Agricultural University, The
Netherlands.
Varnan, A.H., Sutherland, J.P. (1994). Milk and Milk Products. Technology, Chemistry
and Microbiology, Chapman and Hall, London.
Vodickova, M., Forman, L. (1984). Use of buttermilk in quality improvement of ice
cream, Veda a Vyzkum v Potravinarskem Prumyslu, 30, 213-229.
Walstra, P. (1987). Physical principles of emulsion science, in Food Structure and
Behaviour (eds. J.M.V. Blanshard and P. Lillford). Academic, New York, pp.
87-106.
---,Jenness, R. (1984). Dairy Chemistry and Physics, Wiley, New York.
Westerbeck, J.M.M., Prins, A. (1991). Function of a-tending emulsifiers and proteins
in whippable emulsions, in Food Polymers., Gels and Colloids (ed. E. Dickinson),
Royal Society of Chemistry, London, pp. 14 7-158.
Zakharova, N.P., et al. (1979a). The calcium-phosphorus ratio in processed cheese,
210 Food Emulsifiers and Their Applications
Tmdy- Vsesoyuznyi-Nauchno-issledovatel'skii-lnstitut-Maslodel'noi-i-Syrodel'noi-
Promyshlennosti-Nauchno-proizvodstvennogo-Ob "edineniya"-'Uglich,' 27, 105-108,
121. (In Russian)
- - - et al. (1979b). Method for increasing the hydrophilic properties of cheese
mass, 'Tmdy-Vsesoyuznyi-Nauchno-issledovatel'skii-Institut-Maslodel'noi-i-Syrodel'noi-
Promyshlennosti-Nauchno-proizvodstvennogo-Ob"edineniya "-'Uglich,' 27, 108-lll,
121. (In Russian)
Zadow, J.G. (1982). Recombined milks and creams, International Dairy Federation
Bulletin, 142, 33--46.
EIGHT
Applications of Emulsifers
in Baked Foods
Frank T. Orthoefer
8.1 Introduction
The development of emulsifiers or surfactants for bakery products has followed
the development of shortenings. The term "shortening" was initially used to re-
fer to the fats used to "shorten" or tenderize baked foods. The composition of
shortening has progressed from natural fats to blends of oils, hydrogenated
fats, and hard fats. Shortenings may include additives such as emulsifiers, an-
tioxidants, antifoams, and metal scavengers. In addition to their tenderizing
function, the shortening affects structure, stability, flavor, storage quality, eat-
ing characteristics, and eye appeal of baked foods. Many of the functional ef-
fects are due to or enhanced by the emulsifier added via the shortening. This
chapter will concentrate on the role of the emulsifier in baked foods.
century. Initially, cottonseed oil was blended with lard as a "lard compound"
or simply compound shortening. Hydrogenation as a process was invented in
1910. This new process allowed the development of substitutes for the semi-
solid (plastic) animal fats and permitted the creation of products with im-
proved functional properties.
Along with this modification technique came improved methods for pro-
cessing the oil, including refining, bleaching, and deodorization. The products
possessed improved oxidative stability, uniformity, and enhanced perfor-
mance. With the development of lipid chemistry came methods for alcoholysis,
esterification, interesterification, and isomerization. These advances in lipid
chemistry brought new emulsifiers and shortening formulations. High-ratio
shortenings were introduced about 1933. These contained mono- and diglyc-
erides that brought about finer dispersion of fat particles in the dough, giving
strengthened cake batters. These finer particle dispersions allowed higher
sugar levels, with increased water added, to produce sweeter tasting and more
tender cakes. High-ratio shortenings possessed excellent creaming properties.
Moist, higher volume, fine-grained, even-extured cakes were produced. Icings
were improved as well (Hartnett, 1977).
Emulsifier development also proceeded as well in the 1930s (Stauffer, 1996).
Specialty shortenings were the result. Commercial layer cakes, pound cakes,
cake mixes, creme fillings, icings, whipped toppings, bread, and sweet dough
shortenings were created. The development of specialty products resulted in im-
provements in processing and improved product performance for the retail, food
service, and food-processing industries. In addition to the traditional plastic
shortenings, liquid shortenings, fluid shortenings, and powdered products were
produced. All these shortenings involved formulation with emulsifiers.
The terms emulsifiers and emulsifying agents, surfactants and surface-ac-
tive agents are used interchangeably in the literature. The terms "emulsifier"
and "emulsifying agents" are, strictly speaking, chemicals or compounds ca-
pable of promoting emulsification or stabilization of emulsions by their effect
on interfacial tension. Surfactants for foods may include not only emulsifiers
but also compounds with other functions such as protein or starch interaction.
In most cases, the terms are used interchangeably.
The role of the emulsifier and that of the shortening are intimately bound in
bakery products. Generally, the food emulsifiers for bakery products supple-
ment and improve the functionality of a properly developed shortening.
Emulsifiers act as lubricants, emulsify fat in batters, build structure, aerate,
Applications of Emulsifiers in Baked Foods 213
improve eating quality, extend shelf life, modify crystallization, prevent stick-
ing, and retain moisture. A list of emulsifiers commonly used in shortenings is
given in Table 8.1. The selection of or addition of an emulsifier to a shortening
base may significantly change the application of the shortening (Table 8.2).
Nonemulsified Emulsified
All-purpose Cake and icing
Puff pastry Household
Pie crust Filling
Cookie Cake mix
Danish roll-in Yeast raised
Donut fry Specialty cake
Fraction Percent
Starch 70.0-75.0
Protein 11.5-12.5
Pentosan 2.0-2.5
Lipid l.0-1.5
Crude fiber 0.2
Ash 05
OOCCH
I
-QOC-cH-CH-COQ-
Diacetyl-tartaric acid ester of
monoglyceride (DATEM)
1
OOCH 3
Polyoxyethylene monoglyceride or
ethyoxylated monoglyceride
(EMG)
Where
Sorbitan monostearate
and conditioning effects in baked goods. Eight hydroxyl groups that may be es-
terified are present in sucrose. As expected, the degree of esterification affects
the hydrophylic lipophilic balance (HLB) of the sucrose ester (Table 8.4). The
product is considered a noncaloric fat substitute when six or more of the hy-
droxyls are sterified (Olestra™ by Proctor & Gamble).
8.5.5 Lecithin
Lecithin is a coproduct of soybean oil and, to a limited extent, of other veg-
etable oil refining (com oil and rapeseed oil). It is obtained by water-washing
crude vegetable oil and separating and drying the hydrated gums. The crude
material is very dark colored and viscous, consisting of a variety of phospho-
lipids (Figure 8.5). In addition to the phospholipids, these are also present
Applications of Emulsifiers in Baked Foods 221
CH 3
Sodium stearyllactylate (SSL)
{also in calcium form)
0 0
II II _
H3C (CH 2)17-0-c-CH-CH-C-o8NJ±)
OH
I
CH 2-CH-cH 3
I
[0-CH 2-CH-CH 2-OJ n
1
OH
0-H C-HC-CH
2 I 2
OH
Polyglycerol monostearate
0
II
CH 2-0-C-(CH 3 ) 16CH 3
0 0
H
HO
H OH OH H
Sucrose diester
Figure 8.4
8.6.1 Starch
Starch exists in a helical, coiled configuration with about six residues per tum.
This structure is basically a hollow cyclinder that has a hydrophyllic outer sur-
face and a hydrophobic inner core. The inner space is about 45 nm in diame-
ter. Straight-chain, alkyl molecules such as stearic acid will fit in the inner
Applications of Emulsifiers in Baked Foods 223
71 24 5 0 15
61 30 8 1 13
50 36 12 2 ll
46 39 13 2 9.5
42 42 14 2 8
33 49 16 2 6
CH 2-0-R 1
r~~
CH -0-P-Q-R
2 lb 3
a
Phosphatidylethanolamine
Phosphatidyl inositol
Phosphatidylserine
space. The n-alkyl portion of emulsifiers such as present in GMS form a com-
plex with the helical regions of the starch. It is this complex that is thought to
retard starch crystallization in bread, often called "retrogradation," slowing
the staling process.
The influence of emulsifiers on starch consists of (i) affect on the rate of
gelatinization, the gelatinization temperature, the peak viscosity, and the gel
224 Food Emulsifiers and Their Applications
8.6.2 Protein
The wheat flour proteins, gliadin and glutenin, form a viscous, colloidal com-
plex known as "gluten" when mixed into a dough. Lipids are also involved in
the formation of the gluten complex. The properties of the gluten are influ-
enced by the lipids and the emulsifiers present. Lipophilic parts of surfactants
interact with hydrophobic regions of proteins, contributing to unfolding or de-
naturation of the protein. Generally, surfactants contribute to protein denatura-
tion, enhancing interfacial absorption and emulsion stabilization.
Most dough strengtheners are anionic surfactants. The association of the
lipophilic portion to the hydrophobic protein portion incorporates the negative
charge into the complex, promoting aggregation in the dough. The overall ef-
fect is aggregation of the gluten protein and an increase in dough strength.
The ionic surfactants induce protein insolubilization, resulting in increased
viscosity and elasticity of the dough. Nonionic surfactants disrupt the hy-
drophobic portion of the protein, leading to reduced dough viscosity and elas-
ticity and increased protein extractability. A blend of emulsifiers generally
provides the best dispersibility and functionality.
Applications of Emulsifiers in Baked Foods 225
8.6.3 Lipids
Wheat flour contains 1.4 to 2.0% lipids divided into free (0.8 to 1.0%) and
bound (0.6 to 1.0%) forms. They may be further divided into nonpolar (50.9%)
and polar (49.1 %) forms. The bound lipids exist as starch inclusion com-
plexes. The nonstarch lipids, about 85% of the total, participate in the chemi-
cal, physical, and biochemical processes important for the preparation of
baked goods. The nonstarch lipids consist of glycolipids, phospholipids, and
stearyl esters. Interaction between nonstarch lipids and emulsifiers is limited.
Likely, added lipids merely influence or substitute for each other.
Non-polar-lipid addition to untreated flour gives a deterioration of baking
properties (Schuster and Adams, 1984). Addition of polar lipids to untreated
flour increases loaf volume in breadmaking. The improvement is likely based
on the effects of galactolipids and phospholipids. Emulsifiers may interact with
the water phase of the dough, forming associated lipid-water structures with
free polar flour lipids (Krog, 1981). Emulsifiers may compete with naturally
occurring lipids in wheat flour for the reactive groups of the wheat flour dough.
Their effect on protein components was modulated as a consequence.
2900
2800 1-
~l ~
_fl I
00 .10 .25 .50 .10 .25 .50 .10 .25 .50 .10 .25 .50 .10 .25 .50
NO CSL EOM PS-60 SMG SSL
ADD
%Flour, wt
170
160
150
140 I n ~ n
11
r-~ r-J
1
~!I_ _Jl ~J-
j JJ
r---. ; r---
Jl
130
12 o_
- ~- ~ r--- r--- -
NO .10 .25 .50 10.25.50 .10 .25 .50 .10 .25 .50 10.25 .50 .10 .25 .50 .10 .25 .50 .10 .25 .50 1.0
ADD PS-60 SSL SMG Mo-Di LSC Dis. M . H.
CSL EOM
%Flour, wt
Figure 8.7 Relative crumb-softening effect in bread by CSL, EOM, PS-60, SSL, SMG,
Mo-Di (54% mono- and diglyceride), LEC (lecithin), and Dis. M.H. (22% solids dis-
tilled monoglyceride hydrate). (From Tenney, 1978.)
8.7.1.3 Emulsifier Blends. Lecithin has been used in breads and baked
goods longer than any other emulsifier. It was shown to give higher ductility
through interaction with the gluten. Other activity from the lecithin is delayed
staling and shortening reduction. A synergistic effect also occurs with lecithin
and monoglycerides. The monoglycerides produce better crumb grain, softer
bread, and higher loaf volumes. Ethoxylated monoglycerides with monoglyc-
eride is an effective dough conditioner. The negative effects of liquid oils in
place of shortening in bread preparation is overcome with the combination.
DATEM also acts as a dough conditioner, shortening saver, and antistaling
agent in combination with glycerol monostearate. Others include SMG, su-
crose esters, polysorbate 60, SSL, and CSL. The SSL and CSL can form com-
plexes with gluten acting as a dough strengthener.
The direct and indirect action of the emulsifier begins with dough prepara-
tion and ends with oven baking and storage (Figure 8.8). The first stage begins
Applications of Emulsifiers in Baked Foods 229
Improvement of
wettability Decrease of mixing time and
mixing speed
Stabilization of
distributed phases Mixing Reducing shortening levels
Improvement of mixing tolerance
Improvement of machinability
Improvement of gas-retaining
Fermentation properties
Scaling, kneading, Shorter fermentation
moulding Greater shock-tolerance
Improvement of gas-retaining
properties
Improved loaf volume
Baking Better texture
Better crumb grain
Better uniformity
Decrease of water loss
Figure 8.7 Influence of emulsifiers on production and quality of baked products. (From
Schuster arul Adams, 1984.)
with wetting, and dispersing activity then follows with interactions with flour
components during mixing and in the baking process itself.
ide from the leavening agent. The use of emulsifiers lowers the surface tension
of the aqueous phase, improving the amount of air that can be whipped into the
batter. Large amounts of finely divided air cells are important for development
of uniform grain (Randleman et al.,1961). The dissolved C0 2 evolves at air
cell sites and does not spontaneously form bubbles. If the original batter con-
tains many small air cells, the final cake will have a larger volume and fine
(close) grain. The creaming of the sugar and shortening have a major influence
on air incorporation. The incorporation of monoglycerides in the plastic short-
ening (2 to 5% alpha-monoglycerides) ensures numerous small air cells being
created during beating or creaming.
Cake batter is, of course, an aerated emulsion. The integrity of the air cells
determines cake volume and uniformity. Shortening is an antifoam that disrupts
foam cells. Emulsifiers, however, coat the exterior of the fat particles protecting
the integrity of the air cell (Wooten, 1967). Proper emulsifier selection has per-
mitted the use of liquid oil where only shortening could previously be used.
Light, tender, moist cakes are preferred by the consumer. Emulsifiers pro-
vide the desired aeration, emulsification, and crumb-softening. Crumb-soften-
ing in cakes is a function of moisture retention, shortening activity, and
starch-complexing. The starch-complexing is the same as for breads. The
emulsifiers complex with the starch to soften the product.
Several types of emulsifiers are used in cakes. Propylene glycol monoester
(PGME) is utilized at 10 to 15% of the shortening. Monoglycerides and mix-
tures of lactated monoglycerides with PGME are also used in cake mixes.
In bakers' cakes, a large number of emulsifiers are used, depending on the for-
mula, production equipment, and labeling requirements. With soybean oil as the
shortening, a hydrated blend of emulsifiers such as PS 60, SSL, sorbitan mono-
stearate, and distilled monoglycerides work well. Fluid shortenings are produced
containing lactated monoglycerides. The traditional bakers'-cake system is a plas-
tic shortening with 5 to 10% monoglycerides (3% alpha-monoglyceride content).
Packaged cake mixes often use emulsified PGME at 10 to 15% of the oil. The
cakes are unusually tender and are not suitable for commercial cake production.
Emulsified cake shortenings are also used for cake donuts. The degree of air
entrapment during the creaming phase determines the grain in the final donut.
8.7.2.2 Cookies and Crackers. Although emulsifiers are used to only a limited
extent in cookies and crackers, they play a significant role in (i) controlling
spread, (ii) improving cutting and appearance, and (iii) improving texture. The
Applications of Emulsifiers in Baked Foods 231
limited use probably stems from the reluctance to change the successful for-
mulas in this segment of the baking industry. New products utilizing emulsi-
fiers are receiving greater acceptance since they have been shown to play
functional roles.
Certain emulsifiers have the ability to change the degree of spreading of
cookie dough as it is baked (Table 8.5). The effect likely occurs because of al-
tering of the viscosity of the dough. Cookie dough with SSL shows increased
spread compared to a nonemulsified control (Rusch, 1981). The SSL may in-
teract with the starch granule, delaying hydration of the granule and subse-
quent gelatinization (Tsen et al., 1973).
Lecithin may be used to produce a drier dough that machines better and
that improves release from the rotary die surface. Use is from 0.25 to 0.7% of
the flour. Part of the effect may simply be reduction of available water because
of lecithin hydration.
Lecithin that is highly fluidized with other oils or fatty acids is widely used
as release agents in cookie baking due to improved release from rotary dies.
Heat-resistant lecithins such as those modified with acetic anhydride are espe-
cially adaptable to the application. Lecithin is used in cookie and cracker for-
mulations at 0.25 to 1.0% of flour weight. It may be added with the shortening
at the creaming stage or simply combined with the shortening when rotated.
Antistaling is of less significance in cookies and crackers since they are of
Spread
0.5% additive ratio
Monoglyceride 8.3
Ethoxylated monoglyceride 8.8
Sodium stearoyl fumararale 10.4
Sodium stearoyllactylate 10.0
Sucrose monopalmitate 9.8
Sucrose mono- and distearate 9.6
Sucrose distearate 9.7
Sorbitan monostearate 9.2
Polyoxylated (20) sorbitan 9.3
monostearate
Succinylated monoglycerides 9.2
232 Food Emulsifiers and Their Applications
moisture retention and functionality. PGME and DATEM products have as-
sisted in formula development.
Emulsifiers are not generally regarded as fat substitutes or replacers.
Emulsifiers affect the texture and mouthfeel by their effect on surface interac-
tions. The caloric value of emulsifiers varies depending on their actual compo-
sition and digestibility. They tend to have fat-like properties through their
hydration, water-binding, and dispersing effects in processed foods. The gen-
eral functions of emulsifiers in low-fat or no-fat applications are to
Mono- and diglycerides are the most used emulsifiers. Distilled monoglyc-
erides have a caloric advantage over the lower mono-containing preparations.
Other emulsifiers in reduced fat products include the polysorbates, polyglyc-
erol esters, and sorbitan monoesters.
Many noncaloric fats have been synthesized. Sucrose polyesters (OlestraTM
from Proctor & Gamble) is probably the best known. Emulsifier use in prod-
ucts utilizing the noncaloric, synthesized fat replacers is very likely similar to
that utilizing the traditional caloric version.
8.8 Summary
The market for emulsifiers in bakery products continues to grow. Reduced-fat
products have provided the more recent surge in utilization. Producers of
234 Food Emulsifiers and Their Applications
References
Brandt, L. (1996). Emulsifiers in baked goods, Food Product Design, Feb., pp. 64-76.
Chawla, P., deMan, J.M. (1990). ]. Am. Oil Chem. Soc., 67, 329.
Hahn, D.E., Hood, L.F. (1987). Factors influencing com starch lipid complexing, Ger.
Chem., 64,81-85.
Handleman, A.R., et al. (1961). Bubble mechanisms in thick foams and their effect on
cake quality, Ger. Chem., 38, 294.
Hartnett, D.I. (1977). ]. Am. Oil Chem. Soc., 54, 557.
Henry, C. (1995). Monoglycerides: the universal emulsifier, Ger. Foods World, 40(10),
734-738.
Knightly, W:H. (1988). Ger. Foods World, 33,405-412.
Krog, N. (1981). Theoretical aspects of surfactants in relation to their use in breadmak-
ing, Ger. Chem., 58, 158--164.
Lagendijk, J., Pennings, H.J. (1970), Ger. Sci. Today, 15, 354-356, 365.
O'Brien, R.D. (1996). Shortening: types and formulations, in Bailey's Industrial Oil and
Fat Products (ed. Y.H. Hui), 5th ed., Vol. 3, pp. 161-193, Wiley, New York.
Rusch, D.T. (1981). Emulsifiers: uses in cereal and bakery foods, Ger. Foods World,
26(3), 1ll-ll5.
Schmidt, J.C., Orthoefer, F. (1985). Modified lecithins, in Lecithins (eds. B.F. Szuhaj
and G.List), Chap.lO, pp. 203-213, Am. Oil Chern. Soc., Champaign, IL.
Schuster, G., Adams, W.F. (1984). Emulsifiers as additives in bread and fine baked
products, in Advances in Cereal Science and Technology (ed. Y. Pomeranz), Chap.
4, pp. 139-242, Am. Assoc. Cer, Chern. Inc., St. Paul, MN.
Stauffer, C.E. (1996). Emulsifiers for the food industry, in Bailey's Industrial Oil and
Fat Products, 5th ed., Vol. 3, pp. 483-516, Wiley, New York.
Tenney, R.J. (1978). Dough conditioners/bread softeners, Baker's Digest, 52 (4), 24.
Tsen, C.C., et al. (1973). High protein cookies I. Effect of soy fortification and surfac-
tants, Baker's Digest, 47(4); 34-39.
Weiss, T.J. (1983). Food Oils and Their Uses, AVI, Westport, CT.
Wooten, J.C., et al. (1967). The role of emulsifiers in the incorporation of air into layer
cake batter systems, Ger. Chem., 44, 333.
NINE
Emulsifiers in Confectionery
Mark Weyland
9.1 Introduction
The use of emulsifiers in both chocolate confectionery and sugar confectionery
has been established for many years. They are useful functional additives
without which many candy products would be impossible to make. In this
chapter, an outline of the range of emulsifier functionalities will be presented,
but since it is a very diverse field, it is not possible to cover every application
or use. A bibliography is provided for further study, which is recommended.
Emulsifiers act in multiphase confectionery systems in two main ways. The
first is as an emulsifying agent to enable two distinct phases to be combined in
a stable quasi-homogeneous state for an indefinite length of time. The two or
more phases involved in most food products may be selected from a list that in-
cludes oil, water, protein, carbohydrates, and air. These phases have a natural
tendency to repel each other and form discrete layers within a food product. In
most food products this tendency to separate into phases is undesirable and
must be controlled by a suitable blend of processing techniques and carefully
selected emulsifying agents. Furthermore, even if the food product is satisfac-
tory at the time of production, it must still withstand the rigors of distribution
and storage on the shelf, such that at the point of consumption by the con-
235
236 Food Emulsifiers and Their Applications
sumer the product has an acceptable taste, appearance, and texture. These
qualities in modern food products are often critically dependent on the type
and level of emulsifiers used in the product.
The second function of an emulsifier is often to act as a surfactant. In these
cases the role of the emulsifier is to modify the behavior of the continuous
phase of a food product so as to bring about a specific effect or benefit. The
most common example of this in confectionery is the use of lecithin in choco-
late to reduce the viscosity of the product and improve the ease of handling
and processability.
Many of the classes of emulsifiers described in this book have also found
their way into confectionery products. These include lecithin and modified
lecithins such as YN and phosphated monoglycerides, glycerol monostearate,
polyglycerol esters including polyglycerol polyricinoleate (PGPR), sorbitan es-
ters, polysorbates, lactic acid and tartaric acid derivatives of monoglycerides,
acetylated monoglycerides, sucrose esters, and propylene glycol monoesters.
All these compounds have a common feature that makes them suitable as
emulsifying agents: they are ambiphilic, possessing both lipophilic and hy-
drophilic properties. The nature of this property is often expressed as hy-
drophilic/lipophilic balance or HLB. The HLB number is an indication of the
properties of an emulsifier and is usually on a scale of 0 to 20. An emulsifier
with a low HLB will tend to be more oil-like and will therefore have a greater
affinity for the oil phase of a confectionery product. Lecithin, for example, has
an HLB of 4 and has a great affinity for the oil phase in chocolate. Polysorbate
60, by contrast, has an HLB of 15 and is quite soluble in water and therefore
has an affinity for the syrup phase in toffees and caramels.
It is often the case in food products, and in confectionery too, that a combi-
nation of two emulsifiers in a recipe or formula containing two distinct phases
will result in the longer lasting and more uniform product. In these cases, com-
binations oflow- and high-HLB emulsifiers give the best results.
In the remainder of this chapter, a number of the more common confec-
tionery types using emulsifiers are described, along with a review of the avail-
able knowledge relating to the most optimal emulsifier types and their benefits.
9.2.1 Lecithin
Chocolate and compound coatings are dispersions of solid particles in a con-
tinuous fat phase. The solid particles are composed of sugar granules, milk
solids, and cocoa solids. In chocolate the fat is cocoa butter and comes directly
from the crushing of cocoa nibs, but in compound coatings the fat comes from
vegetable oils added to the formula to give the same fat content as chocolate
overall. Moisture is also present in the coating, introduced indirectly via the
sugar or other solid ingredients. It is the presence of these solid particles and
moisture that causes chocolate and compound coatings to deviate from true
Newtonian viscosity behavior. The solid particles tend to abrade each other,
and the resultant internal friction causes the viscosity of the material to vary
according to the applied shear stress.
Viscosity is very important when considering how chocolate and compound
coatings are used, because they always have to flow to either fill a mold without
defects or air bubbles or cover a candy piece with a thin, even coat. These
properties are controlled almost completely by the rheological behavior of the
coating, which in turn is dependent on the nature of the continuous liquid
phase, that is, the fat and fat-soluble ingredients. Considering the flow proper-
ties of molten chocolate, the two most important parameters are plastic viscos-
ity and yield value. "Plastic viscosity" is defined as the force required to keep
liquid chocolate flowing once it has started moving, whereas "yield value" is
the force required to start the mass of liquid chocolate moving. Both plastic
viscosity and yield value can be decreased by the use of specific surfactants,
and this enables the chocolate manufacturer to have greater control of cocoa
butter levels. Plastic viscosity and yield value are often combined in a value
called "apparent viscosity," but it is important to understand that chocolates
with equal apparent viscosities can have different yield values and different
plastic viscosities.
238 Food Emulsifiers and Their Applications
Coatings can always be made more fluid for better control by adding more
cocoa butter or vegetable fat to the mix, but since these are the more costly in-
gredients in coatings, this is often an unattractive solution. Better by far is to
add a surfactant like lecithin to reduce coating viscosity. It is possible to add
0.5% lecithin to a coating to have the same viscosity reduction effect as the
addition of 5% cocoa butter or vegetable fat (Minifie, 1980). Lecithin allows
coating users to operate efficiently at much lower fat contents than would oth-
erwise be the case (Figure 9-1).
Lecithin is commercially extracted from the soybean by solvent extraction
and precipitation. It is a light brown fluid that contains approximately 65%
acetone-insoluble phosphatides and 35% soybean oil. An oil-free product is
also available that is in the form of granules. The chemical composition of soy
lecithin is as follows (Minifie, 1980):
39
38
L.
Q) 37 1\
~
::I
.0 36
\
co
""
0
(.)
0 35
~
(.)
'#- 34
-
..........
33
['... ~
32
0.1 0.2 0.3 0.4 0.5 0.6 0.7
% lecithin (soya)
Figure 9.1 Effect of addition of lecithin on the fat content in dark enrobing chocolate.
Curve represents chocolate formulas of equal viscosity. (Minifie, 1980.)
Emulsifiers in Confectionery 239
0
I
CH 2-o-c-R
lipophile
I ?!
CH-0-C-R
0 hydrophilic
I II
CH 2-o-P-O-cH 2-CH 2-N-(CH 3 ) 3
I II
OH OH
100
[',_
1'--- I I
90 ~ r--__
~Cocoa/Fat
70 c--·- --
!\
\\
--: 60
"'utJ
~ \ -·· - -
..
50 t---- -
\l
\
'
>.
-
0
u 40 Chocolate
I .........
>"' I'-.
30
20
10 "" I
1'--.:_ugor I Fat
0 .1 0 .2 0 .3 0.4 0.5 0 .6 0 .7
"I• Lecithin (Soya)
process in the fat phase. Levels of over l% in typical 30% fat coatings should
not be used.
a neutral flavor and can be used at higher dosage levels than natural lecithin
without the negative impact on viscosity (Bonekamp-Nasser, 1992; Klienert,
1976; Nakanishi, 1971). See Figure 9.4.
YN is also claimed to reduce the thickening of chocolate and compound
coatings due to moisture and overheating (Bradford, 1976). A comparison be-
tween lecithin at 0.3%, YN at 0.3%, and cocoa butter added at 5% to choco-
late gave the same similar overall viscosity readings (see Figure 9.5), but a
calculation of the Casson yield values showed that YN produced significantly
lower values than with the other systems. The viscosity reducing effect of YN
is reportedly less in milk-free coatings than with milk coatings (Klienert,
1976; Hogenbirk, 1989). Milk coatings have generally higher viscosities than
milk-free coatings due to the effect of milk solids/fat/emulsifier interactions.
150
~ 1 RP.M.
-
140
130 ~\ ---·YN
Soya lecithin
-
120 ~\ RPM. : Rate of shear
-
\\ r--------
\\
110 --
100 --
....
\"''
""
~ 90 --- - --
g
tl ao ' --
0"' ..............
?- 70 ''
.....
r--
~ ..........
... ... ...
--.. _
"'u0 60
>"' 50
'-..5 R.P.M. ...
"0
u 40
'~
~
0 '<"..... r::--..........
... -r--
-
0
L. 30
..........
.... --
cD
20
--- 1-----
20 R.P.M. ~
-------
10 ~--
0
0.1 0.2 0.3 04 0.5
0 /o Lee ith 1 n
-
700
c:;- 5% cocoa butter
E
..s?.
Ul
600
Q)
c: 0.3% lecithin
> 500
~ --+-
Ul
Ul 0.3% YN
....
Q)
+'
Ul
400 __.._
....
ctl
Q) 300
..c:
(/)
200
0 10 20 30 40 50
Shear rate (s-1)
Figure 9.5 Viscosity plot comparing lecithin, YN, and cocoa butter. (Bradford, 1976.)
where R= H or a fatty acyl group derived from poly condensed ricinoleic acid
and n is the degree of polymerization of glycerol.
Emulsifiers in Confectionery 243
A number of studies have been published that compare the effects of PGPR
with lecithin and YN. Most conclude that PGPR, when added to chocolate or
compound coatings at 0.5% or less, can reduce the coating yield value to al-
most zero (Application Notes Admul WOL, Quest International; Bamford,
1970). The practical benefit of such a feature is that in a chocolate bar molding
operation, PGPR addition would allow the chocolate to flow easily into even
complicated mold shapes without entrapping air bubbles and also to flow
around inclusions. Furthermore, the opportunity exists to reduce the fat con-
tent of the chocolate as well as the coat of chocolate formulations.
A typical comparison of lecithin and PGPR additions to a milk and to a
dark chocolate using a Rotovisco viscometer is given in Tables 9.1 and 9.2
(Application Notes Admul WOL, Quest International).
The values in Tables 9.1 and 9.2 indicate that in milk chocolate it is possi-
ble to reduce the yield value to almost zero. The presence of lecithin in the
emulsifier mix allows the plastic viscosity to be minimized, and this blend pro-
Table 9.1 Casson plastic viscosities and yield values of a milk chocolate when
cocoa butter, lecithin, and PGPR are added
Table 9.2 Casson plastic viscosities and yield values of a dark choco-
late when cocoa butter, lecithin and PGPR are added
vides the optimum solution. In dark or semisweet chocolate the effect of PGPR
on plastic viscosity is small, while it can reduce yield values to very low values
at 0.5% addition. The mechanism by which PGPR affects yield value is not
understood but its practical benefits are widely reported.
PGPR is also claimed to be advantageously used in ice cream coatings since
it allows low apparent viscosities in the presence of low levels of moisture
(Bamford, 1970). Also claimed is PGPR's beneficial effect on fat-phase crystal-
lization leading to easier tempering, improved texture, and longer shelflife of
coatings (Application Notes Admul WOL, Quest International.). The author has
found that the viscosity-reducing properties of PGPR does lead to significantly
reduced viscosity at temper and a level of temper, as measured by a temperme-
ter, which is easier to maintain over long periods in an enrober without signifi-
cant recirculation of chocolate via melt-out and retempering circuits.
PGPR's most recognized benefit remains that of fat reduction, and manufac-
turers claim that a blend of 0.5% lecithin and 0.2% PGPR allows cocoa butter
reductions of approximately 8%.
ter-based chocolate is to ensure that the cocoa butter crystallizes in the cor-
rect crystal form or polymorph. Cocoa butter has several different polymorphic
forms that have melting points ranging from 17 to 35°C. The forms are repre-
sented by the greek letters y, a, pt, and~· As the polymorphic form increases
in stability it also increases in melting point. To make chocolate in the familiar
glossy, fast-melting form with good snap, it is necessary to crystallize the cocoa
butter in the highest melting polymorph, which is ~· This form of cocoa butter
is also needed to ensure good contraction in molded products and the long
bloom-free shelf life expected for good quality chocolate goods. Bloom is the
phenomenon in which liquid fat is forced to the surface of a product and spon-
taneously crystallizes there as a grayish layer that looks to the uninformed con-
sumer as if the product has gone moldy. The liquid fat is forced out to the
surface in this way by the following mechanisms:
sociated with bloom in solid chocolate that has been well tempered. Work done
by Garti et al. (1986) has indicated that STS is particularly effective at block-
ing this V to VI transformation and, hence, preventing bloom even after exten-
sive temperature cycling between 20 and 30°C.
Other emulsifiers studied by Garti included sorbitan monostearate and
Polysorbate 60, but these were only half as effective as STS. STS is a high-
melting-point emulsifier (approximately 35°C} whose structure is more closely
related to cocoa butter triglycerides than to most other emulsifier types. It is
speculated that it is due to this similarity that STS co-crystallizes with cocoa
butter from the melt and, due to its rigid structure, binds the lattice in form V.
Other more liquid or less triglyceride-like emulsifiers tend to depress the melt
point of crystallized cocoa butter, increasing liquidity and promoting form IV
to V transformations in preference.
This is presumably why STS is a more effective antibloom agent in solid
chocolate than in enrobed chocolate items, where soft center oils can soften
cocoa butter crystals via migration and lead to type IV-V bloom occurring (see
type 3 bloom previously mentioned). Krog (1987), however, claims that STS
locks fats in the less stable Wform and prevents the transformation to ~
Berger (1990) also claims that STS performs well as a bloom inhibitor or gloss
enhancer in palm kernel oil-based compound coatings used to enrobe cakes
by stabilizing the Wform of the vegetable fat, a situation also observed by the
author in several practical cases using lauric coating fats but with much less
reliability when using domestic fats such as soybean or cottonseed-based coat-
ing fats. Such products tend to have longer bloom-free shelf lives in many
cases so that the need for antibloom additives is not so imperative.
The use of STS in food in the United States is limited to chocolate and com-
pound coatings for which there is a petition pending for acceptance by FDA as
a food additive. This uncertain status has not deterred United States food com-
panies from enjoying the benefits that STS can bring to coating appearance.
STS is, however, more widely accepted as an additive in EC countries.
giving them a lighter texture for filling applications. Herzing (1982) describes
in detail the types of polyglycerol esters needed to optimize the glossy proper-
ties of lauric and nonlauric compound coatings: triglycerol monostearate, oc-
taglycerol monostearate, and octaglycerol monooleate. These emulsifiers are
added to the coating fat at up to 6% by weight.
Polyglycerol esters have also been claimed to speed up the setting time of
chocolate panning coatings when used at levels of 0.4 to 0.6% (Player, 1986).
Hogenbirk (1989) has found advantages of viscosity reduction to some degree
with examples of mono- and diglycerides, diacetyl tartaric acid esters of mono-
glycerides (DATEM), acetylated monoglycerides, and propylene glycol mo-
noesters. Musser (1980) has published results showing the benefits of adding
up to 1.5% DATEM to chocolate and compound coatings to modify viscosity
properties and to improve the rate of crystallization of coating fat phases. In a
series of experiments, Musser found that the addition of DATEM to fully
lecithinated milk and dark chocolates, and dark sweet coatings, could further
reduce the viscosity of the coatings as measured by a Brookfield viscometer. At
the same time Musser found that the DATEM was acting as a seeding agent in
chocolate systems, improving the speed of crystallization and resulting in a
finer grain and better gloss in molded bars. Musser's conclusions relative to the
effect of DATEM on viscosity is supported by the author's own published study
on chocolate viscosity and emulsifiers (Weyland, 1994).
References
Anon. (199la). Confectionery Production, 57(2), 136-7, 140.
- - - (l991b). Confectionery Production, 57(6), 451-2.
Bamford, H.F., eta!. (1970). Rev. Int. Choc. (RIC), 25, 6.
Berger, K.G. (1990). World Conference on Oleochemicals into the 21st century, AOCS,
Champaign, IL, pp. 288-91.
Bonekamp-Nasser, A. (1992). Confectionery Production, 58(1), 66, 68.
Bradford, L. (l976).Int. Flavors Food Addit., 7(4), 177-9.
Dave, ].C., eta!. (1991). U.S. Patent 5,135,761, March 28.
Dziezak, J.D. (1988). Food Techno!., 42(10), 171-86.
Garti, N., eta!. (1986). ]. Assoc. Off. Chem. Soc., 63(2) 230-236.
Gregory, D.H. (1982). Confectionery Production, 48(10), 437-9.
Harris, T.L. (1968). Surface Active Lipids in Foods, Monograph No. 32, Society of
Chemical Industry, England.
Herzing, A.G., eta!. (1982). U.S. Patent 4,464,411, November 5.
Hogenbirk, G. (1989). Confectionery Production, 55(1), 82-3.
Jeffrey, M.S. (1991). Manufacturing Confectioner, 71(6), 76-82.
Klienert, J. (1976). Rheol. Texture Food Qual., pp. 445-73, AVI, Westport, CT.
Krog, N. (1977). ]. Assoc. Off. Chem. Soc., 54(3), 124--31.
Lang, M. (1974). Confectionery Manufacturing and Marketing, 11(2), 3-5, 13.
Lees, R. (1975). Confectionery Production, 41(6), 296,298, 304.
254 Food Emulsifiers and Their Applications
10.1 Introduction
The invention of margarine in the 1860s is attributed to the French chemist
Hippolyte Mege Mouries, in response to a competitive challenge initiated by
the French Government under Napoleon III for a less expensive and less per-
ishable substitute for butter.
Mege had already undertaken research in this field at the Imperial farms,
where he had observed that although underfed or hungry cows lost weight, they
still produced milk, though of lower than normal yield, and that the milk con-
tained fat. Thus, he deduced that milk fat was derived from normal body fat,
i.e., tallow. However, since tallow itself does not possess the melting property
of milk fat, he construed that through some metabolic process, a fractionation
of the fat occurred, the lower melting fractions of which were transported to the
mammary glands of the udder whereby, through the enzymatic action of
pepsin, it became transformed to butter fat and dispersed as an emulsion in
the milk plasma, while the harder fractions were utilized by the animal as a
source of energy (Andersen and Williams, 1954).
Having arrived at these conclusions, Mege set out to imitate this natural
process by carefully rendering fresh tallow at body heat (about 45°C) using ar-
255
256 Food Emulsifiers and Their Applications
tificial gastric juices to facilitate separation of tissue from pure fat and then,
following crystallization at 25 to 30°C, extracting under pressure about 60% of
a soft semifluid fraction-oleomargarine-and about 40% of a hard white
fat-oleostearine. The softer fraction had a melting point similar to butter fat,
a pleasant flavor not unlike melted butter fat, a pale yellow color, and could
easily be plasticized, and although different in fatty acid composition from but-
ter fat, it offered a useful basis for the production of a substitute. He assumed,
therefore, that this new butter fat substitute consisted of glycerides of margaric
and oleic acids (the former then considered to be a C17 homolog of the satu-
rated fatty acids, although now known to be a eutectic mixture of palmitic and
stearic acids) that solidified in crystals having a pearly luster. The name mar-
garine was derived from the Greek word "margarites," meaning pearl and thus
is pronounced with a hard "g" according to its derivation.
Mege's process was to take a proportion of the soft fat with appropriate
quantities of milk and water, into which a small amount of udder extract was
stirred. Following agitation, a stable emulsion similar to thick butter cream
formed that, on further churning, thickened to resemble butter. While this
process may now sound rather rudimentary, it nonetheless comprised all the
elements of margarine production as we know them today.
A similar development has been seen in Europe (Table 10.2), where in re-
cent years (and also forecast to continue) the trend is for a reduction in yellow-
fat consumption generally, with small reductions in both butter and margarine
in the order of 4 to 5% and an increase in spreads of 17 to 19% by 1997.
be similar to butter. However, with the new developments during more recent
years involving different components and varying combinations of vegetable
oils, animal fats, and milk fat, a range of new descriptions have come into be-
ing such as low-fat spreads, low-calorie spreads, and yellow-fat spreads. In
1993, Moran listed the varieties then available, as shown in Table 10.3.
Approx. fat
content(%)
A. Milk fats l. Butter Milk-fat content not less than 80% but less than 90%.
Products derived exclusively from milk Max. water 16%; max. dry nonfat milk materials 2%
and/or certain milk products for which fat 2. Three-quarter-fat butter Milk-fat content not less than 60% but not more than 62%
is the essential constituent of value 3. Half-fat butter Milk-fat content not less than 39% but not more than 41%
4. Dairy spread xo/o Products with milk-fat content:
• Less than 39%
• More than 41 %; less than 60%
• More than 62%; less than 80%
B. Fats l. Margarine Vegetable and/or animal fats not less than 80%; less than 90%
Products derived from solid and/or liquid 2. Three-quarter-fat Not less than 60%; less than 62%
vegetable and/or animal fats suitable margarine
for human consumption, with a milk-fat 3. Half-fat margarine Not less than 39%; more than 41%
content more than 3% of the fat content 4. Fat spreads xo/o Products with fat content:
• Less than 39%
• More than 41 %; less than 60%
• More than 62%; less than 80%
C. Fats composed of plant and/or l. Blend Product from a mixture of vegetable and/or animal fats
animal products with fat content of not less than 80% but not more than 90%
Products derived from solid and/or liquid 2. Three-quarter-fat blend Not less than 60%; not more than 62%
vegetable and/or animal fats suitable for 3. Half-fat blend Not less than 30%; more than 41%
human consumption, with a milk-fat 4. Blended spread xo/o Products with fat content:
content of between 10 and 80% but not more than • Less than 39%
• More than 41 %; less than 60%
• More than 62%; less than 80%
Whether the listed descriptions will gain greater prominence other than as
the required secondary descriptions is doubtful since consumers, presumably,
will prefer to favor the brand names that have been applied to the broad range
of products now available.
Fat spreads falling outside these standards, for instance those with fat con-
tents below 10% and concentrated products with fat contents of 90% or more,
will not be classed as spreadable fats.
EC legislation does not specify vitamin fortification, which is left for deci-
sion at the national level. However, in the UK the requirement will remain as
previously specified in the Margarine Regulations (Sl 1867) 1967. That is,
each 100 g of margarine shall contain 800 to 1000 mg of vitamin A and 7.05 to
8.82 mg of vitamin D. This does not apply to other spreads, so that vitamin for-
tification in these cases is a matter for individual decision by the manufactur-
ers and/or retailers.
Legislation in the United States and Canada is currently less detailed than
in Europe, but both require a minimum of 80% fat. In the United States, FDA
§156.110 describes margarine (or oleomargarine) as "food in plastic form or
liquid emulsion" containing not less than 80% fat and describes a method for
fat determination. It specifies vitamin A fortification to be not less than 15,000
I.U. per pound but leaves vitamin D as an optional ingredient while stating not
less than 1500 I.U. per pound. While spreads are not specified in the legisla-
tion, low-fat spreads were stated in 1990 as the fastest growing category, with
spreads from 20 to 72% fat on the market (Borwanker and Buliga, 1989).
The National Association of Margarine Manufacturers of the United States
describes spreads as having fat contents ranging from 18 to 79%. Details of
market shares in the United States are shown in Table 10.5.
Stick margarine 65 44
Soft margarine 19 l3
Liquid margarine 1 2
Low-fat vegetable oil spreads 15 41
Total retail market 100 100
where R 1, R2 , and R3 are the fatty acid residues, which may be the same but,
invariably, are different according to the source of the fat. Examples of the
variations in fatty acid compositions of different oils and fats are shown in
Table. 10.6.
Modification of the fat by, for instance, hydrogenation, whereby some of the
unsaturated links (double bonds) become saturated (for instance oleic C18:1 to
stearic CIS) offer the opportunity of considerable variation in the melting
points of fats, especially the liquid vegetable oils. The exact physical character
of a fat depends upon that of the constituent fatty acids that can vary widely in
melting point, as illustrated in Table 10.7.
""' Low-
Beef Ground- Palm erucic Sun-
Fatty acid tallow Lard Coconut nut Palm kernel rapeseed Soya flower
C-10 & lower; - - - 13-17
capric, etc.
C12lauric 0.2 0.2 45.9-50.3 - tr 43.6-51.4 - tr
C14 myristic 3-4 1.6 16.8-19.0 tr 0.8-1.3 15.3 17.2 tr tr tr
C14:1 myristoleic 0.5-1 tr - - tr
C16 palmitic 26-28 25-30 7.7-9.7 9.2-13.9 43.1-46.3 7.2-10.0 3.4-6 9.9-12.2 5.6-7.4
C16:1 palmitoleic 2-3 2.5-3.5 - tr tr - 0.2-0.6 tr tr
C18 stearic 23-27 15-25 2.3-3.2 2.2-4.4 4.0-5.5 1.9-3.0 l.l-2.5 3.6-5.4 3.0-6.3
C18:1 oleic 30-35 35-45 5.4-7.4 36.6-65.3 36.6-65.3 11.9-18.5 52-65.7 17.7-25.5 14.0-34.0
C18:2 linoleic 1-1.5 7-12 1.3-2.1 15.6-40.7 9.4-11.9 1.4-3.3 16.9-24.8 50.5-56.8 55.5-73.9
C18:3 linolenic 0-1 0.5-1 tr tr 0.1-0.4 tr 6.5-14.1 55-95 tr
C20 & higher 4-8 1-4 tr 2-8 [xx]1 [xx]1 1-6 [xx]l.5 [xx]l.5
Iodine valuet 45-57 59-70 7.5-10 84-105 50-54 16-19 109-126 125-136 85
tiodine value is a measure of the degree of unsaturation of the fat.
Margarines and Spreads 265
Coconut 3 20
Coconut/palm 1:1 4 15
Coconut/palm, interesterified 5 18
Palm-hardened 5 17
Palm-hardened/palm 2:1 8 13
Lard 14 lO
Lard/palm 1:1 15 10
Palm 27 lO
Sheafat 45 10
mediate, and the most stable ~ form the highest values. Transition from one form
to another is from a ~ W~ ~ and is not reversible without remelting the fat.
The polymorphism of crystalline fats may cause problems with the consistency
of margarine and spreads. During production, the fats initially crystallize in the a
form and normally will rapidly transform to the Wform. This is the desirable form
for spread production since the small needle-shaped Wcrystals (about l!lm long)
impart good plasticity. Should the crystals transform from Wform to the much
larger p form (> 20 Jlm) they will give the spread a grainy consistency known as
"sandiness," as can be seen in Figure 10.1 (Madsen and Als, 1968).
266 Food Emulsifiers and Their Applications
(a) (b)
Figure 10.1 Fat crystals in (a) normal margarine with good consistency and (b) a mar-
garine with large ~crystals causing a "sandy" texture. (Magnification 200x.) (Courtesy
of Danisco Ingredients, Denmark.)
Concurrently, the specific area of the crystal surface will decrease, allowing
the liquid oil to penetrate to the surface of the margarine, which may then lead
to oiling out, especially if the margarine comes under pressure. Some veg-
etable oils such as partially hardened sunflower or low erucic rapeseed are
particularly prone to form ~ crystals and thus can cause sandiness in spreads.
In this case, sorbitan tristearate at 0.3% of the fat has been found to inhibit the
transition from the Wto the ~ form
The crystal-modifying effect of a range of emulsifiers-sorbitan esters,
ethoxylated sorbitan esters, ethoxylated fatty alcohols, citric acid esters of
monoglycerides (Citrem), diacetyl tartaric acid esters of monoglycerides
(DATEM), sucrose monostearate, sodium stearoyl lactylate, and polyglycerol
esters- has been investigated on the polymorphism of tristearin (glyceryl tris-
tearate) (Garti et al., 1982).
In these trials it was found that sorbitan monostearate and Citrem (Cl6/Cl8
fatty acids) were the most effective in preventing the recrystallisation (from a
to ~ form) of tristearin.
It should be noted that when used in emulsions, surface-active materials will
Margarines and Spreads 26 7
be adsorbed at the oil/water interface and that only lipophilic emulsifiers with
high solubility in the oil phase can perform as crystal inhibitors in emulsions.
Varying the level of saturated fat affects the degree of crystallinity of the oil
phase (Borwanker and Buliga, 1989). Spreads with 40% fat were made using
oil phases of varying crystallinity by blending either liquid soyabean oil or
stick margarine oil with soft margarine oil. The textures of the spreads made
with these blends were visibly very different, increasing in softness in the
same direction as the oil itself. As a consequence of increased firmness, the
stability of the resulting spread was expected to increase with increasing crys-
talinity in the oil phase. A centrifugation procedure was used to measure the
stability of the emulsions (Figure 10.2). The higher amount of water and/or oil
(especially water) released corresponds to a less stable emulsion.
10.7 Emulsifiers
The main function of emulsifiers in processed foods is to reduce the interfacial
tension between the phases of an emulsion-usually oil and water. In such
two-phase systems, one phase is dispersed as large droplets within the other.
60
+-'Cil
c
Cll co
u Cll
....
"0
Ul
-
50
O Water
.s-.:
"-~
Cll Cll
30
>.0 -a...___
..~I...
==o 20
..c.._
co Cll ~
+-'+J
CJ)co 10 ~
3:
0
4.5 6.3 8.6 10.4 13.0
Figure 10.2 Effect of the degree of crystallinity on the emulsion stability of 40% fat
spreads as measured by ultracentrifugation. (From Borovanker and Buliga, 1989.)
268 Food Emulsifiers and Their Applications
They are either oil-in-water (0/W) emulsions, where the continuous phase is
water (such as in milk or ice cream) or water-in-oil (W/0), where the continu-
ous phase is oil (as in butter and margarine).
The stability of two-liquid phase emulsions is kinetic stability, i.e., the system
is not thermodynamically stable (Friberg et al., 1990). Thermodynamically sta-
ble emulsions would spontaneously reform following separation by, for instance,
centrifugation, while experience shows that an emulsion that has separated re-
mains so unless mixed by some external action. In reality, the two separated
phases are in the most stable state to which all emulsions will tend. Thus, a sta-
ble emulsion is one in which this inevitable trend has been retarded so that it is
not noticeable during the normal life of the product--even if it be several years.
Margarine is a water-in-oil emulsion in principle only because, in reality, it is
a dispersion of water droplets in a semisolid fat phase, containing fat crystals
and liquid oil (Krog et al., 1983). Preparation of the emulsion requires consider-
able energy to reduce the droplet size of the disperse phase, thereby creating an
increase in the surface area between the two immiscible phases.
The initial water-in-oil emulsion is prepared in mixing tanks with vertical or
horizontal stirrers to ensure emulsification but without incorporating too much air.
It is rather coarse in water-droplet size and fairly unstable if not kept agitated.
In modern continuous production, the emulsion exists for only a brief pe-
riod before passing into the chilling unit where final emulsification and crys-
tallization of the fat phase take place. Thus, the emulsion need not be very
stable against coalescence as the water droplets become fixed within the semi-
solid phase. However, the droplet size is important when considering flavor re-
lease and microbiological spoilage. Thus, emulsifiers are used to lower the
interfacial tension between the oil and water phases, which will generally re-
sult in a smaller water-droplet size. Distribution of the water droplets in the
range 2 to 4 11m will give better stability against deterioration such as mold
growth. However, it is still desirable for some of the water droplets to be larger
in size-10 to 20 11m-to give a better flavor release in the mouth.
For these purposes, therefore, lipophilic emulsifiers such as mono-/diglyc-
erides of long-chain fatty acids (C16-C18) are used at levels of 0.1 to 0.3%,
often in combination with 0.05 to 0.1% refined soya lecithin.
Comparisons of the water-droplet distribution in margarine emulsion and in
finished margarine are shown in Figure 10.3. Droplet size in the emulsion is
further reduced during the cooling and kneading processes in the tube chiller.
The water droplets in finished margarine are stabilized by adsorbed fat
Margarines and Spreads 269
(a) (b)
Figure 10.3 Water-droplet size distribution in (a) margarine emulsion and (b) the fin-
ished margarine. (Magnification 200x.) (Courtesy of Danisco Ingredients, Denmark. )
Figure 10.4 Freeze fracture electron micrograph showing the microstructure of mar-
garine. Water droplets (w) are covered with fat crystals. (Bar: l!lm.) (Courtesy of Dr. W.
Buchheim, Keil.)
10.8 Processing
Typical modern processing involves the separate processing of the oil and wa-
ter phases. The oils and fats are kept in storage tanks at temperatures just ade-
quate to maintain them at the required fluidity. Preferably, they should be
bottom fed to avoid splashing and aeration. It is essential that the plant, i.e.,
tanks, pipes, and pumps, be entirely free from copper or copper alloys to avoid
the high risk of oxidation.
The final oil blend together with oil-soluble components such as emulsi-
fiers, colors, flavors, and vitamins, is prepared separately from the water phase
Margarines and Spreads 271
that may contain milk components such as whey powder, skim milk powder, or
sodium caseinate and also salt, gelatin, or thickener, water-phase flavors where
appropriate, and preservatives such as potassium sorbate for low-fat products.
Modern methods involve the feeding of the phases by computer-controlled
load cells, which ensures fast throughput and constant composition.
A second emulsion tank is used for maintaining quantities of finished emul-
sion for feeding the cooling and kneading equipment. Cooling and kneading
can be carried out by either of two different methods: (i) tube chiller or (ii)
chilling drum-complector.
In the former, cooling and kneading take place in a closed system and in a
single process. In the latter, the cooling and kneading are carried out sepa-
rately-cooling on the chilling drum and kneading in the complector. The ad-
vantage of the chilling drum-complector method is that it allows the product to
rest between cooling and kneading, which is important with formulas based on
slower crystallizing fats, for instance, puff pastry margarine. By comparison,
the tube chiller is relatively more compact considering throughput, is easy to
operate, and reduces the possibility of spoilage.
Most types of margarine can be made very satisfactorily using the tube
chiller method, although in some cases extra working units (pinning machines)
may prove advantageous. However, the chilling drum-complector has been the
preferred method for puff pastry margarine, although tube chillers are now
· widely used, and is perfectly satisfactory for normal table margarine and for
cake margarine. The suitability of both methods is summarized in Table 10.9.
Chilling
drum-
Type Tube chiller complector
The comparative merits of the two methods were investigated using four differ-
ent fat combinations (see Table 10.10) in puff pastry margarines that were then
evaluated by various parameters including a "finger" judgement (see Table
10.11) of plasticity after one week (Madsen, TPlOl).
Table 10.10 Dilatation, iodine value, and melting points of fat blends used for
comparison of puff pastry margarines
tThe figures indicate storage and rolling temperature of the puff pastry margarine.
Source: Madsen (fPlOl).
Similar results were found when the same products were judged after 3
weeks' storage.
This rather critical evaluation did not fully take into account the differ-
ences that might result from the use of varying techniques possible with tube
chillers, which was investigated separately.
Table 10.12 Plasticity judgment by means of the finger method in puff pastry
margarines made on tube chiller with varying production techniques
The important issue relates to the postcrystallization of the fats, which was
further investigated (Madsen, 1981 ). A good plastic puff pastry margarine can
be bent without breaking and the plasticity evaluated by repeated handwork-
ing to check stability, firmness, and greasiness (Figure 10.5).
In the case of cake margarine, the most important points are that the fat
blend is soft and easy to incorporate into the batter and has good creaming
properties. This, therefore, suggests the use of lauric fats (coconut, palm ker-
nel) that crystallize quickly and, thereby, facilitate creaming.
274 Food Emulsifiers and Their Applications
of the fat blend, the type and level of emulsifier, and the addition of thickeners
such as gelatin, sodium alginate, pectin, and carrageenan to the aqueous phase.
Low levels of whey protein may be used to improve flavor release with the further
advantage that the pH of the aqueous phase can be lowered to improve keeping
properties because, unlike casein, whey proteins do not precipitate at low pH.
The speed of processing, i.e., the rate of throughput and the emulsion tempera-
ture, are also important factors in the stability of the spread.
Examples of the effect of various types of emulsifiers on the stability of low-
fat spreads with fat contents of 40% and 20% are shown in Tables 10.13 and
10.14, respectively.
Table10.13 Effect of types and blends of emulsifiers on the stability of a 40% fat
spread
IV = Iodine value.
Formulation of spread:
Water phase 56.4% water
0.5% whey powder
pH 4.5 1.5% gelatin
1.5% salt
0.1% potassium sorbate
Fat phase 39.2-39.4% fat blend
0.{H).8% emulsifier (as above)
4 ppm beta-carotene
IV = Iodine value.
Formulation of spread:
The trial batches in both cases were made on a three-tube Perfector pilot
plant using a fat blend composed of
Both examples indicate the potential instability of these emulsions and the
variation in the stabilizing effects of different emulsifier blends even when used
at low dosage levels. These results suggest that emulsifier structure at the inter-
face plays a critical role in retarding water-droplet flocculation and coalescence.
While the evaluation of spreads by measuring water separation provides
quantifiable results, a quicker and more practical evaluation of the stability of
Margarines and Spreads 277
However, in this case it is not possible to lower the pH of the aqueous phase
without precipitating the caseinate, which will reduce its emulsifying proper-
ties, and therefore, the keeping properties of the spread will be limited.
However, a satisfactory low-fat butter spread can be produced from dairy
cream, using a distilled monoglyceride with high iodine value (80 to 105) in
the cream and a thickener such as sodium alginate. In this case, phase inver-
sion from 0/W to W/0 can be achieved in the tube chiller, using normal or
slightly reduced cooling and operating at 40 to 50% of the usual capacity. To
obtain a satisfactory working effect, a high rotor speed in the tube chiller cool-
ing cylinder would be preferable (Madsen, 1989).
Factors that may affect the efficiency of phase inversion in low-fat emul-
sions are listed in Table 10.15.
278 Food Emulsifiers and Their Applications
Most of these factors concern the increase in rate of collision of oil droplets
upon which the rate of coalescence is dependent (Moran, 1993). The emulsi-
fier(s) likely provide steric hindrance to keep droplets separated.
Against this, however, one major drawback is that unless products are pre-
pared at comparatively low pH, then ultra-high-temperature processing and
possibly an aseptic filling procedure must be followed if shelf lives comparable
to conventional spreads are required.
Oil-in-water spreads remain a relatively unexplored area of the spreadable
fats market, possibly due to the problem of microbiological deterioration, but
may offer a potential for future expansion in markets where adequate levels of
preservatives are permitted. In contrast to W/0 emulsions, these systems
would utilize high-HLB emulsifiers to stabilize the water-continuous emulsion.
In markets such as the UK that have traditionally used solid fats like lard or
cooking fat or, more recently, liquid vegetable oils, there is less interest in us-
ing liquid margarine.
A typical composition has a fat phase of about 82% based on soya or sun-
flower oil, in which emulsifiers perform two functions. First, an emulsifier such
as citric acid ester of monoglyceride (Citrem) at, say, 0.4% plus 0.2% soya
lecithin produces a stable water dispersion with limited spattering during fry-
ing; and second, a selected mono- and diglyceride blend prevents oiling out on
storage and thus gives a homogenous product with low viscosity. If the mar-
garine is not intended for frying, the Citrem can be replaced with 0.2% dis-
tilled monoglyceride. The water phase may contain proportions of skimmed
milk powder and salt as well as a preservative such as potassium sorbate.
Flavoring may be added to both phases.
The aqueous phase should be adjusted to 4.5 to 6.5 pH and pasteurized at
80°C. The fat phase should be tempered to about 60°C and the emulsifiers
melted into a small amount of the liquid oil to a temperature of 65 to 70°C,
which is then stirred into the main part of the fat phase. The water phase is
added to the fat phase under continuous agitation before cooling through the
tube chiller with an outlet temperature of approximately 5 to 7°C.
The emulsion should be rested for 2 hours to allow proper fat crystal forma-
tion and then stirred vigorously for 15 minutes before topping off for packaging.
10.12 Summary
Standard margarines containing 80% fat are, under normal conditions, rela-
tively stable, requiring minimal quantities of lecithin and/or mono- and diglyc-
erides over and above the quantities of milk proteins usually present.
However, where the margarine is required for special purposes, i.e., cake-
making or frying and particularly in products with reduced fat levels, emulsi-
fiers specific to the unique functional requirements of the application should
be selected (see Chapter 8 for applications of emulsifiers in baking products).
References
Andersen, A.J.C., Williams, P.N. (1954). Margarine, 2d ed., Pergamon, Oxford.
Borwanker, R.P., Buliga, G.S (1989). Emulsion properties of margarines and low fat
spreads, in Proc. Food Emulsions and Foams-Theory and Practice (eds. P.G. Wan
eta!.), San Francisco, pp. 44-52.
280 Food Emulsifiers and Their Applications
Flack, E. (1992). The role of emulsifiers in reduced fat and fat free foods, in Food
Technology International Europe (ed. A. Turner), Sterling Publications, London, pp.
179-181.
Friberg, S.E. et al. (1990). Emulsion stability, in Food Emulsions (eds. K. Larrson and
S.E. Friberg), Marcel Dekker, New York, pp. 1-40.
Garti, N., et al. (1982).]. Am. Oil Chem. Soc., 59, 181.
Krog, N., et al. (1988). Applications in the food industry, in Encyclopaedia of Emulsion
Technology, Vol. 2: Applications of Emulsions (ed. P. Becher), Marcel Dekker, New
York, p. 321.
Madsen, J. Puff Pastry Margarine: A Comparison of the Chilling Drum and Tube Chiller
Methods of Production, TP101, Grinds ted, Denmark.
- - - (1981). Post-Crystallisation in Puff Pastry Margarine, at 11th Scandinavian
Symposium on Lipids, Bergen, Norway.
- - - (1983). Product Formulation and Processing of Margarine and Yellow Fat
Spreads, at Margarine and Yellow Fat Seminar, Coventry, England.
- - - (1989). Low Calorie Spread and Melange Production in Europe, at World
Conference on Edible Oils and Fats Processing, Maastricht, Holland.
- - - , Als, G. (1968). Sandiness in Table Margarine and the Influence of Various
Blends of Triglycerides and Emulsifiers Thereon, at /Xth Int. Soc. of Fat Research
Congress, Rotterdam.
Moran, D.P.J. (1993). Reduced calorie spreads, in Porim Technology, No. 15., Feb '93,
Palm Oil Research Institute of Malaysia.
Schwitzer, M.K. (1956). Margarine and Other Food Fats, Leonard Hill, London.
ELEVEN
Emulsifier Trends
for the Future
Gerard L. Hasenhuettl
part of the physical structural or primary functionality of food. For example, the
development of high liquid-oil margarines will require an emulsifier that can sta-
bilize a W/0 emulsion where the continuous phase is liquid. Adjunct technolo-
gies, such as the development of molecular networks to thicken the oil phase can
make a significant contribution to providing structure to the processed food.
Solid fat is implicated in such functions as aeration of whipped toppings and for-
mation of evenly dispersed air cells in cakes. Replacement with liquid oil places
more of a functional burden on the emulsifier system.
Health authorities have recommended that total dietary fat should not ex-
ceed 30% of calories. Some consensus is also building that some forms of can-
cer may correlate to total caloric consumption. Since fat represents 9 cal/gram,
cutting fat consumption will also reduce total calories (if the same weight of
food is consumed). If consumers respond to these recommendations, pressures
for availability of reduced-fat and fat-free foods will continue or even increase.
Elimination of fats from products normally containing high levels creates enor-
mous challenges to maintain texture and flavor. Flack (1992) has speculated
that emulsifiers can play an important role in reclaiming some of these quali-
ties. Structured emulsifier phases have been patented as the basis for forma-
tion of a pseudo-fat phase (Heertje, et al., 1994; Kleinherenhrink et al., 1995).
This trend toward replacement of fat functionality with emulsifiers will un-
doubtedly continue.
very efficient surfactants for application such as enhanced oil recovery and oil-
spill control. A logical extension of these technologies would be to develop
food-grade organisms that produce natural surfactants. However, extensive
evaluation would still be essential to ensure the safety of these products when
consumed in processed foods.
Interfacial
(hydrophobic)
attraction , . - - / H e a d-group (hydrophilic) repulsion
~
Volume v ·-._ \
Area a 0
the functional role of a mesophase is defined, rational design of head and tail
groups can he accomplished and the emulsifier synthesized by techniques de-
scribed in Chapter 2.
Emulsifiers may he viewed as one class of functional food ingredients. As
Chapters 4 and 5 have pointed out, these ingredients may interact with carbo-
hydrates and proteins to produce a number of technical effects. The area of in-
gredient interactions is the subject of a recent book by Gaonkar (1995). New
experimental techniques such as scanning tunneling microscopy and interfa-
cial rheology have the potential to provide greater understanding of these in-
teractions. Emulsifiers can then be designed to form more selective interactive
structures that may result in improved or entirely new functionalities.
References
Arcos, J.A., Otero, C. (1996). ]. Am. Oil Chem. Soc., 73:673-82.
Flack, E. (1992). Food Techrwlogy International-Europe, 79-81.
Gaonkar, A. (ed.) (1995). Ingredient Interactions: Effects on Food Quality, Marcel
Dekker, New York.
Heertje, 1., et al. (1994). European Patent 0 558 523 Bl, July 13.
lsraelachvili, J.N. (1985). Intermolecular and Surface Forces: With Applications to
Colloidal and Biological Systems, Academic, New York, pp. 246--62.
Kleinherenbrink, F.A., et al. (1995). International Patent Application WO 95/35035,
December 28.
Larsson, K. (1994). Lipids-Molecular Organization. Physical Functions and Technical
Applications, The Oily Press, Ayr, Scotland, pp. 156--62.
Shah, D.O., et al. (1994). ]. Am. Oil Chem. Soc., 71:1405-9.
Vulfsen, E.N. (1993). Trends Food Sci. & Techno!., 4:209.
Index
Index
c PGPR, 242-244
polysorbate 60, 24 7
cake batters, 212, 230 sorbitan:
calcium stearoyl-2-lactylate, 20, 57 tritearate, STS, 246--24 7
calcium hydroxide, 17 monostearate, 24 7
capillary melting point, 53 synthetic lecithin, 240
Index 291
separations of phospholipids, 58 J
homogenization, 166
jellies, 251
hydrated distilled monoglycerides, 227
ACI values, 252
hydrodynamic interactions, 166
hydrophile/lipophile balance (HLB),
6-8,151,163-164,169,
220,236 K
hydrophilic, 5, 11, 100
kosher, 5, 13
hydrated lime, 17
Krafft temperature, 148
hydration:
force, emulsion droplets, 155
repulsion, monopalmitin, 155
L
lactic acid, 236
analyses of, 49
ice cream, 95, 174-183 water-in-soluble combined,
aging of, 178 (WICLA), 22
composition of, 175 esters of:
coalesence stability of, 176 glycerol, 22
cryo scanning of, 177 monoglycerides, 187, 248
emulsifiers, 175, 178 propylene glycol, 22
fat globles in, 179 lactalbumin, 102, 123, 125, 130, 185
fat crystallization in, 180-182 lactoglobulin, 102, 104, 106, 109,
interfacial viscosity, 177 113-114,117, 118,
manufacture of, 175 120-121,122,124,125,
orthokinetic stability, 177 126,130,133,134,185
inclusion complexes, starch, 68 lactylated esters, 18-23, 41
interesterification process, 17 hydroxyl group reaction, 19
interfacial tension, role, 165-166 fatty acids of, 19
in bakery products, 217 metal salts of, 19, 20
iodine preparation of, 19-23
dilation value, margarine, 272 lactylated monoglycerides, 20-22,
melting point, fat blends, 272 23,44,227,230
value, 45--46 preparation of, 20-22
binding of starches, 77 lamellar liquid-crystalline phase,
ionic surfactants, 100 emulsifiers of, 153
isotherm adsorption, 100 lamellar phase, monoglycerides, 154
isotherm surface tension, 115 Langmuir-Blodgett technique, 121
IUPAC, 56 layer adsorption, 167, 169, 171
296 Index
y
v yeast-raised products, 225
van der Waals interaction, 98 crumb-softening, 225--227
vesicle bilayers, 131 dough conditioning, 225
viscosity, 8, 54,177,237,241-244 , dough strenghthening, 225, 226
248 emulsifier functionality during
apparent, 237 processing, 227
interfacial, 177 yield value, 237
plastic, 237, 243, 244 yoghurt, 202