Nutritional Evaluation of Processed Pigeon Pea and Roasted Pumpkin Seeds

NUTRITIONAL EVALUATION OF PROCESSED
PIGEON PEA AND ROASTED PUMPKIN SEEDS
BY
Rania Abbas Ahmed Abusham

B. Sc (Agric. Honours)
University of Khartoum – Faculty of Agriculture
A dissertation submitted to University of Khartoum in partial

fulfillment for the requirements of the degree of Master of
Science in Food Science and Technology
SUPERVISOR
Prof. Elfadil Elfadl Babiker
DEPARTMENT OF FOOD SCIENCE AND

TECHNOLOGY
FACULTY OF AGRICULTURE
UNIVERSITY OF KHARTOUM
May 2007
DEDICATION
To my family especially my parents
to my dear friends
to those whom I will never forget.
i
ACKNOWLEDGMENTS
I would like to express my deepest thanks to my supervisor Prof.
Elfadil Elfadl Babiker for supporting, advising and encouraging me
throughout the course of my study, and Thanks for his personal guideness
and fruitful criticism from which I benefit much.
My deep gratitude to all staff members of the Department of
Food Science and Technology, Faculty of Agriculture, University of
Khartoum, for their help.
My thanks to my family, friends and colleagues, who were ready
to render any assistance I ask for to complete this work.
ii
ABSTRACT
The effect of dehulling, cooking and cooking followed by dehulling of
pigeon pea seeds and the effect of roasting of pumpkin seeds on the
proximate composition, antinutritional factors, protein availability,
protein fractions and digestibility of major fractions were investigated.
Processing method of pigeon pea seeds were observed to reduce
moisture, ash, oil and fiber content. However, dehulling of pigeon pea
seeds increased protein content. Roasting of pumpkin seeds increased oil
and fiber content, on the other hand, decreased moisture, ash and protein
content. All processing methods applied were observed to reduce tannins
and polyphenols content of both pigeon pea and pumpkin seeds.
Dehulling of pigeon pea seeds significantly (p ≤ 0.05) increased phytic
acid content from 158.44mg/100g to 173.16mg/100g. However, other
treatments applied reduce phytic acid content for pigeon pea and pumpkin
seeds. All processing methods applied were observed to improve protein
digestibility for both pigeon pea and pumpkin. Cooking of pigeon pea
seeds followed by dehulling was observed to increase protein digestibility
from 61.25% to 91.25%, as a high level obtained compared to other
treatments. Dehulling of pigeon pea seeds significantly (p ≤ 0.05)
increased albumin and globulin fractions. Cooking and cooking followed
by dehulling significantly (p ≤ 0.05) reduced albumin and globulin
fractions. However, all processing methods had no significant (p ≤
0.05) effect on prolamin and glutelin fractions. On the other hand,
roasting of pumpkin seeds significantly (p ≤ 0.05) reduced Albumin,
globulin and prolamin fractions. Glutelin significantly (p ≤ 0.05)
increased as a result of roasting. Processing methods applied for both
pigeon pea and pumpkin seeds significantly (p ≤ 0.05) increased albumin
and globulin digestibility.
iii
‫ﺒﺴﻡ ﺍﷲ ﺍﻟﺭﺤﻤﻥ ﺍﻟﺭﺤﻴﻡ‬
‫ﺍﻟﺨﻼﺼﻪ‬
‫ﺘﻡ ﺩﺭﺍﺴﺔ ﺘﺄﺜﻴﺭ ﻋﻤﻠﻴﺔ ﺍﻟﺘﻘﺸﻴﺭ‪ ،‬ﺍﻟﻁﺒﺦ ﻭﺍﻟﻁﺒﺦ ﻤﺘﺒﻭﻉ ﺒﺎﻟﺘﻘﺸﻴﺭ ﻟﺒﺫﺭﺓ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﻭﺘﺄﺜﻴﺭﻋﻤﻠﻴﺔ‬
‫ﺍﻟﺘﺤﻤﻴﺹ ﻟﺒﺫﺭﺓ ﺍﻟﻘﺭﻉ ﻋﻠﻰ ﺍﻟﺘﺭﻜﻴﺏ ﺍﻟﺘﻘﺭﻴﺒﻰ‪ ،‬ﻤﻀﺎﺩﺍﺕ ﺍﻟﺘﻐﺫﻴﺔ‪ ،‬ﺍﻟﺒﺭﻭﺘﻴﻨﺎﺕ ﺍﻟﻤﺘﺎﺤﻪ‪ ،‬ﺘﺠﺫﺌـﺔ‬
‫ﺍﻟﺒﺭﻭﺘﻴﻥ ﻭﻤﻌﺎﻤل ﻫﻀﻡ ﺠﺯﺌﻰ ﺍﻟﺒﺭﻭﺘﻴﻥ ﺍﻷﺴﺎﺴـﻴﻥ )ﺍﻻﻟﺒﻴـﻭﻤﻴﻥ‪ ،‬ﺍﻟﻘﻠﻭﺒﻴـﻭﻟﻴﻥ(‪ .‬ﻟـﻭﺤﻅ ﺃﻥ‬
‫ﺍﻟﻌﻤﻠﻴﺎﺕ ﺍﻟﺘﻰ ﺃﺠﺭﻴﺕ ﻋﻠﻰ ﺒﺫﺭﺓ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﺃﺩﺕ ﺍﻟﻰ ﺍﻨﺨﻔﺎﺽ ﻓﻰ ﻤﺤﺘﻭﻯ ﻜل ﻤﻥ ﺍﻟﺭﻁﻭﺒﻪ‪،‬‬
‫ﺍﻟﺭﻤﺎﺩ‪ ،‬ﺍﻟﺯﻴﺕ ﻭﺍﻻﻟﻴﺎﻑ‪ ،‬ﺍﻻ ﺃﻥ ﺍﻟﺘﻘﺸﻴﺭ ﺃﺩﻯ ﺍﻟﻰ ﺯﻴﺎﺩﻩ ﻓﻰ ﻤﺤﺘﻭﻯ ﺍﻟﺒﺭﻭﺘﻴﻥ‪ .‬ﺘﺤﻤـﻴﺹ ﺒـﺫﺭﺓ‬
‫ﺍﻟﻘﺭﻉ ﺃﺩﻯ ﺍﻟﻰ ﺯﻴﺎﺩﻩ ﻓﻰ ﻤﺤﺘﻭﻯ ﻤﺴﺘﺨﻠﺹ ﺍﻻﻴﺜﺭ ﻭﺍﻻﻟﻴﺎﻑ ﻭﺍﻨﺨﻔﺎﺽ ﻓﻰ ﻤﺤﺘﻭﻯ ﺍﻟﺭﻁﻭﺒـﻪ‪،‬‬
‫ﺍﻟﺭﻤﺎﺩ ﻭﺍﻟﺒﺭﻭﺘﻴﻥ‪ .‬ﺃﻅﻬﺭﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﻤﻌﻅﻡ ﺍﻟﻤﻌﺎﻤﻼﺕ ﺍﻟﺘﻰ ﺃﺠﺭﻴﺕ ﺃﺩﺕ ﺍﻟﻰ ﺍﻨﺨﻔﺎﺽ ﻤﻌﻨﻭﻯ‬
‫ﻓﻰ ﻤﺤﺘﻭﻯ ﺍﻟﺘﺎﻨﻴﻨﺎﺕ ﻭﺤﻤﺽ ﺍﻟﻔﻴﺘﻴﻙ ﻭﺍﻟﺒﻭﻟﻰ ﻓﻴﻨﻭﻻﺕ ﻤﻘﺎﺭﻨﺔ ﺒﺎﻟﻌﻴﻨﻪ ﻏﻴﺭ ﺍﻟﻤﻌﺎﻤﻠﺔ ﻟﻜـل ﻤـﻥ‬
‫ﺒﺫﺭﺘﻰ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﻭﺍﻟﻘﺭﻉ‪ ،‬ﺍﻻ ﺃﻨﻪ ﻤﻥ ﺍﻟﻤﻼﺤﻅ ﺃﻥ ﻋﻤﻠﻴﺔ ﺍﻟﺘﻘﺸﻴﺭ ﻟﺒﺫﺭﺓ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴـﻰ ﺃﺩﺕ‬
‫ﺍﻟﻰ ﺫﻴﺎﺩﺓ ﻤﻌﻨﻭﻴـﻪ ﻓـﻰ ﺤﻤـﺽ ﺍﻟﻔﻴﺘـﻙ ﻤـﻥ ‪158.44‬ﻤﻠﺠـﻡ‪100/‬ﺠـﺭﺍﻡ ﺍﻟـﻰ ‪173.16‬‬
‫ﻤﻠﺠﻡ‪100/‬ﺠﺭﺍﻡ‪ .‬ﺃﻤﺎ ﺒﻘﻴﺔ ﺍﻟﻤﻌﺎﻤﻼﺕ ﺍﻟﺘﻰ ﺃﺠﺭﻴﺕ ﺃﺩﺕ ﺍﻟﻰ ﺍﻨﺨﻔﺎﺽ ﻓﻰ ﻤﺤﺘﻭﻯ ﺤﻤﺽ ﺍﻟﻔﻴﺘﻴﻙ‬
‫ﻟﻜل ﻤﻥ ﺒﺫﺭﺘﻰ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﻭﺍﻟﻘﺭﻉ‪ .‬ﺃﻅﻬﺭﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﺠﻤﻴﻊ ﺍﻟﻤﻌﺎﻤﻼﺕ ﺍﻟﺘﻰ ﺃﺠﺭﻴﺕ ﻋﻠﻰ‬
‫ﺒﺫﺭﺘﻰ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﻭﺍﻟﻘﺭﻉ ﺃﺩﺕ ﺍﻟﻰ ﺯﻴﺎﺩﻩ ﻓﻰ ﻤﻌﺎﻤل ﻫﻀﻡ ﺍﻟﺒﺭﻭﺘﻴﻥ‪ ،‬ﻜﻤﺎ ﻟﻭﺤﻅ ﺃﻥ ﺍﻟﻁـﺒﺦ‬
‫ﻤﺘﺒﻭﻉ ﺒﺎﻟﺘﻘﺸﻴﺭ ﺍﺩﻯ ﺍﻟﻰ ﺯﻴﺎﺩﻩ ﻜﺒﻴﺭﻩ ﻓﻰ ﻤﻌﺎﻤل ﻫﻀﻡ ﺍﻟﺒﺭﻭﺘﻴﻥ ﻟﺒﺫﺭﺓ ﺍﻟﻠﻭﺒـﺎ ﺍﻟﻌﺩﺴـﻰ ﻤـﻥ‬
‫‪ %61.25‬ﺍﻟﻰ ‪ %91.25‬ﻜﺄﻋﻠﻰ ﻨﺴﺒﻪ ﻟﻬﺎ ﻤﻘﺎﺭﻨﺔ ﺒﺒﻘﻴﺔ ﺍﻟﻤﻌﺎﻤﻼﺕ‪ .‬ﺘﻘﺸﻴﺭ ﺒﺫﺭﺓ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ‬
‫ﺃﺩﻯ ﺍﻟﻰ ﺯﻴﺎﺩﺓ ﻤﻌﻨﻭﻴﺔ ﻟﻸﻟﺒﻴﻭﻤﻴﻨﺎﺕ ﻭﺍﻟﻘﻠﻭﺒﻴﻭﻟﻴﻨﺎﺕ‪ ،‬ﺒﻴﻨﻤﺎ ﺍﻟﻁﺒﺦ ﻭﺍﻟﻁﺒﺦ ﻤﺘﺒﻭﻉ ﺒﺎﻟﺘﻘـﺸﻴﺭ ﺃﺩﻯ‬
‫ﺍﻟﻰ ﺍﻨﺨﻔﺎﺽ ﻤﻌﻨﻭﻯ ﻓﻰ ﻤﺤﺘﻭﺍﻫﻤﺎ‪ ،‬ﻜﻤﺎ ﺃﻨﻪ ﻟﻴﺱ ﻫﻨﺎﻟﻙ ﺍﻯ ﺘـﺄﺜﻴﺭ ﻤﻌﻨـﻭﻯ ﻋﻠـﻰ ﻤﺤﺘـﻭﻯ‬
‫ﺍﻟﺒﺭﻭﻻﻤﻴﻥ ﻭﺍﻟﻘﻠﻭﺘﻴﻠﻴﻥ‪ .‬ﻟﻭﺤﻅ ﻤﻥ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﻋﻤﻠﻴﺔ ﺘﺤﻤﻴﺹ ﺒﺫﺭﺓ ﺍﻟﻘﺭﻉ ﺃﺩﺕ ﺍﻟﻰ ﺍﻨﺨﻔـﺎﺽ‬
‫ﻤﻌﻨﻭﻯ ﻓﻰ ﺠﻤﻴﻊ ﻤﻜﻭﻨﺎﺕ ﺍﻟﺒﺭﻭﺘﻴﻥ ﻤﺎﻋﺩﺍ ﺍﻟﻘﻠﻭﺘﻠﻴﻥ ﺤﻴﺙ ﺃﻅﻬﺭ ﺯﻴﺎﺩﻩ ﻤﻌﻨﻭﻴﻪ ﻜﻨﺘﻴﺠﻪ ﻟﻠﻤﻌﺎﻤﻠﻪ‪.‬‬
‫ﺠﻤﻴﻊ ﺍﻟﻤﻌﺎﻤﻼﺕ ﺍﻟﺘﻰ ﺃﺠﺭﻴﺕ ﺃﺩﺕ ﺍﻟﻰ ﺯﻴﺎﺩﻩ ﻤﻌﻨﻭﻴﻪ ﻓﻰ ﻤﻌﺎﻤل ﻫﻀﻡ ﺍﻻﻟﺒﻴﻭﻤﻴﻥ ﻭﺍﻟﻘﻠﻭﺒﻴـﻭﻟﻴﻥ‬
‫ﻟﻜل ﻤﻥ ﺒﺫﺭﺘﻰ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﻭﺍﻟﻘﺭﻉ‪.‬‬

LIST OF CONTENTS
Page
DEDICATION................................................................................. …..i
ACKNOWLEDGEMENTS...................................................................ii
ABSTRACT .......................................................................................... iii
ARABIC ABSTRACT........................................................................... iv
LIST OF CONTENT............................................................................ v
LIST OF TABLES........................................................................... viii
LIST OF FIGURES............................................................................... ix
CHAPTER ONE: INTRODUCTION …………………………1

CHAPTER TWO: LITERATURE REVIEW ……………….. 4
2.1 Legumes as source of proteins ……………………………….….. 4
2.2 The importance of oil seeds as a source of protein ……………….. 5
2.3 Pumpkin seeds as a source of oil and protein ………………….. 5
2.4 Chemical composition of pigeon pea and pumpkin seeds …............6
2.4.1 Moisture Content ………………………………………………… 6
4.2 Crude protein content ………………………………………………. 7
2.4.3 Fat content ……………………………………………….8
2.4.4 Crude fiber content …………………………………………….9
2.4.5 Ash content …………………………………………………….9
2.4.6 Carbohydrate content …………………………………………….10
2.5 The nutritional value of seeds ………………………….. ………….10
2.6 Effect of processing on nutrients …………………………………...11
2.7 Antinutritional factors …………………………………………… 12
2.7.1 Phytic acid of pigeon pea and pumpkin seeds ……………………13
2.7.2 Tannins content of pigeon pea and pumpkin seeds… ...... ………14
2.7.3 Polyphenols of pigeon pea and pumpkin seeds…… ……………..15
2.8 Effect of processing on antinutritional factors …………………….15
2.9 In vitro protein digestibility (IVPD) of pigeon pea and
Pumpkin seeds ……………………………………………………16
2.10 Effect of processing on In vitro protein digestibility (IVPD) …..17
2.11 Protein quality of seeds ………………………………………….18
2.12 Protein fractions of pigeon pea and pumpkin seeds ………………19
2.12.1 Albumins and globulins ……………………………………. 19
2.12.2 Prolamin ………………………………………………………. 20
2.12.3 Glutelin …………………………………………………………21
2.13 Effect of processing on protein fractions ………………………….21
2.14 Digestibility of protein major fractions …………………………..21
‫اﻏﺎل‬
CHAPTER THREE: MATERIALS AND METHODS................... 24
3.1 Materials......................................................................................... 24
3-2 Methods ………………………………………………………… . 24
3.2.1 Preparation of samples …………………………………………... 24
3.2.2 Defatting of pigeon pea and pumpkin seeds ……………………..24
3.2.3 Analytical methods ………………………………………………25
3.2.3.1 Proximate analysis of samples …………………………………25
3.2.3.2. Determination of anti-nutritional factors ……………………. 25
3.2.3.2.1. Determination of tannin content …………………………… 25
3.2.3.2.2. Determination of phytic acid ………………………………. 26
3.2.3.2.3 Determination of polyphenols ………………………………. 27
3.2.3.3 In vitro protein digestibility determination ……………………27
3.2.4 Physiochemical properties ……………………………………….28
3.2.4.1 Protein fractionation due to solubility ……………………… 28
3.2.4.1.1 Determination of water-soluble proteins ……………………..28
3.2.4.1.2 Determination of salt soluble proteins ……………………….29
3.2.4.1.3 Determination of alcohol soluble proteins ………………..29
3.2.4.1.4 Determination of alkali-soluble proteins …………………… 30
3.2.4.1.5 Protein content of insoluble fraction ……………………….30
3.2.5 Statistical analysis ………………………………………………..30
CHAPTER FOUR: RESULTS AND DISCUSSION …………… .31

4.1 Effect of processing on the proximate composition of defatted
pigeon pea seeds …………………………………………………..31
4.2 Effect of roasting on the proximate composition of defatted
pumpkin seeds ………………………………………………32
4.3 Effect of processing on antinutritional factors of defatted pigeon pea
seeds ………………………………………………………………..34
4.4 Effect of roasting on antinutritional factors of defatted pumpkin
seeds ……………………………………………………………….35
4.5 Effect of processing on in vitro protein digestibility (IVPD) of
defatted pigeon pea seeds …………………………………………35
4.4 Effect of roasting on in vitro protein digestibility (IVPD) of defatted
pumpkin seeds …………………………………………………37
4.7 Effect of processing on protein fractions of defatted pigeon pea
seeds ……………………………………………………………...37
4.8 Effect of roasting on protein fractions of defatted pumpkin seeds …40
4.9 Effect of processing on digestibility of protein major fractions of
defatted pigeon pea seeds ……………………………………… 42
4.10 Effect of roasting on digestibility of protein major fractions of
defatted pumpkin seeds …………………………………………..42
CONCLOSION................................................................................ 45
RECOMMENDATIONS.............................................................. ..... 45
REFERENCES................................................................................... 46
LIST OF TABLES
Page
Table 1. The proximate composition of defatted pigeon pea and pumpkin
seeds …………………………………………………… ……….. 33
Table 2. Antinutritional factors of defatted pigeon pea and pumpkin seed…...36
Table 3. Protein fractions of defatted pigeon pea and pumpkin seeds ……….41
Table 4. Digestibility of defatted pigeon pea and pumpkin seeds protein major
Fractions ……………………………………………………………44
Viii
LIST OF FIGURES
Page
Fig 1. in vitro protein digestibility (IVPD) of processed
pigeon pea seeds …………………………………………. …38
Fig 2. in vitro protein digestibility (IVPD) of roasted
pumpkin seeds ……………………………………………… …..39
ix
NUTRITIONAL EVALUATION OF PROCESSED
PIGEON PEA AND ROASTED PUMPKIN SEEDS
BY
Rania Abbas Ahmed Abusham

B. Sc (Agric. Honours)
University of Khartoum – Faculty of Agriculture
A dissertation submitted to University of Khartoum in partial

fulfillment for the requirements of the degree of Master of
Science in Food Science and Technology
SUPERVISOR
Prof. Elfadil Elfadl Babiker
DEPARTMENT OF FOOD SCIENCE AND

TECHNOLOGY
FACULTY OF AGRICULTURE
UNIVERSITY OF KHARTOUM
May 2007
CHAPTER ONE
INTRODUCTION
Legumes are valuable agricultural crops because of their capacity to

provide a grain crop and also fix nitrogen to soils and improve the soil
value for further cropping Gladstones (1998). Legumes are of great
nutritional importance in Africa. It is needless to emphasize that legumes
can contribute greatly to the improvement of human and animal nutrition,
as well as fertility of the soil. Food legumes are important for a
nutritionally well balanced diet, specially for lower income people Salih
(1992). Pigeon pea (Cajanus cajan (L.) Mill sp.) is the fifth most
important pulse crop in the world. Pigeon pea is an important grain
legume commonly grown and consumed in tropical and subtropical
regions of the world. The plants grow to maturity in six months on high
ground with abundant rainfall. India accounts for over 80% of the worlds
supply of pigeon pea (ICRISAT 1986). Other countries where pigeon pea
is an important food legume are Keanya, Malawi, Uaganda, Tailand,
Indonesia, and Philppines. Pigeon pea provides valuable sources of
protein for people in developing countries, particularly among those law-
income groups who depend on vegetarian diets for economic and social
reasons. Morton (1976) listed many folk medical uses of pigeon pea. In
India and Java, the young leaves are applied to sores. Scorched seed,
added to coffee, are said to alleviate headache and vertigo. Fresh seeds
are said to help incontinence of urine in males, immature fruits are
believed of use in liver and kidney ailments. Green pods of pigeon pea
also harvested and the developing green seed are used as a vegetable in
India and some African, Latin American and south–east Asian countries
Ali (1996). Pigeon pea is a multipurpose crop that can provide food, fuel
wood and fodder in subsistence agriculture, it also supplies calories,
vitamins, and minerals. It entrances soil fertility and its deep rooting
1
pattern enables it to exploit available moisture at greater soil depth, its
multipurpose and uses make it attractive and economical to the resource-
poor farmer Ali (1996). On the other hand, Pumpkin belongs to the
family cucurbitaceous, most of which are tendril climbing herbaceous
annuals containing some extremely well known edible fruits such as
pumpkin, squash, cucumber, musk-melon, and cucurbita toxana. Little
information is available concerning pumpkin grown in Sudan and no
research has been carried as far as we know. There are no distinct
varieties or known species. There are many wild varieties that differ
according to the areas where they are grown. The varieties in Sudan
probably belong to Cucurbita moschata (Qara assally) as stated by
Ahmed (1985). Most of pumpkins consumed in Sudan are grown in
Kordofan (Western-Sudan) where it contributes up to 39.1% total
vegetables and fruits produced. Like other members of cucurbitaceous,
pumpkin fruits bear numerous seeds, where 50 to 70% of the full mature
fruit consists of rind and seed. Beside its use in production of edible oil,
pumpkin seeds where recommended in the literature as a protection
against colworm, tapeworm and interruption of pregnancy and extracted
oil has been put to medicinal use by physicians all over the world
Markovic and Bastric (1976). In Sudan pumpkin seeds are partially
utilized by direct consumption but large quantities are wasted. In western
Sudan some people use pumpkin seeds as anti-diarrheas agent after being
kept in youghourt for a period of time (personal communication). The
availability of pumpkin seeds and the high potential of the seed as oil
seed promote study of seed composition, oil characteristics and its
stability. Cirrilli (1971) reported that pumpkin scoured seeds were
employed for human feeding after previous salting and roasting
processes. In Sudan pumpkin seeds are considered as a by product. Little
amounts are eaten after being salted and roasted (tasali), or exported
2
together with water-melon seeds to Egypt. Recently increased attention
has been given to the utilization of agriculture of secondary products to
produce food, feed, fertilizer and a raw material in certain industries,
which help to maximize the available resources and at the same time to
minimize waste disposal problem. Such utilization could be done
economically only in the locations where such resources are available in
large quantity.
Objectives:
The objectives of the present study are
1- To study the nutritional value of treated (dehulling, cooking and
cooking followed by dehulling) pigeon pea seeds.
2- To study the nutritional value of roasted pumpkin seeds.
3
CHAPTER TWO
LITERATURE REVIEW
2.1 Legumes as source of proteins:

Plant protein play a significant role in human nutrition; they are specially
needed in developing countries Hassan (1994). Kakade and Lay (1974)
reported that the protein nutritive value may be defined as the ability of a
protein to provide a pattern of amino acids in proper concentrations
similar to body proteins depending upon factors such as protein
concentration, protein quality and protein digestibility. Ahmed and Nour
(1990) stated that leguminous seeds play small role in the Sudanese diet
until people recently started to consider them as part of their diet due to
the escalating prices of animal products. The protein content of essential
selected Sudanese leguminous seeds is high: faba bean (29%), cow pea
Spp. (24-26%), pigeon pea (22%) and soy bean (38%). George and
Delumen (1991) reported that legumes are the richest sources of proteins
among plant food but are deficient in the sulfur amino acids methionine
and cysteine. Johnson and Lay (1974) showed that cysteine is not an
essential amino acid but can partly replace dietary requirements for
methionine. Mosse (1990) stated that cereal grain and legumes or oil
seeds protein used for human food and for animal feed. It is thus,
important to have an accurate knowledge of the protein concentration of
grain. This becomes more indispensable in many areas: in human or
animal nutrition, in marketing grain according to protein content and in
the feed industry. In Sudan as in most tropical countries little work has
been carried out either on composition or cultivation of legume crops. It
is well known that the proteins in cereals and millets which form the
major component of poor mans diet are deficient in lysine and that they
4
can be supplemented by legumes which are rich not only in lysine but
also in threonine Venkat Rao et al.,(1964).
2.2 The importance of oil seeds as a source of protein:

Proteins are unquestionably, the most important of all known substances
in the organic and soft tissue of human body, though a liberal and
continuous supply is need in the food throughout life. Proteins are
essential food components and are a source of energy and amino acids.
Most of the plant proteins commercially available are obtained from
cereal, oil seeds, and pulses Dimler (1971). During the last decades, oil
seeds have attracted great interest as a potential source of protein for
human consumption Gheyasuddin et al., (1970). This interest is due to
serious protein deficiencies in some parts of the world and to increasing
costs of many food ingredients form animal sources Sosulski and Mc
Cleary (1972). Accordingly considerable research is oriented to explore
and focus on new protein sources. Since the production of animal protein
(meat, fish, egg, milk, etc) is more expensive. Holm and Breedon (1983)
reported that the use of vegetable proteins as additives or components in
foods have increased markedly in recent years
2.3 Pumpkin seeds as a source of oil and protein:

Edible oil extracted from pumpkin seeds has been highly appreciated and
considered very good for health Bastic et al., (1977). A few workers
found that the pumpkin seeds are good source of protein and minerals
such as iron and phosphorous. In many developing countries, the supply
of animal protein is inadequate to meet the protein needs of the rapidly
growing population. The seeds also are considered as a good source of
oil where Datta (1977) found that pumpkin seeds contain higher oil
content value (51.1%) within the family cucurbitaceous. The protein of
5
fluted pumpkin seed is markedly deficient in the sulphur–containing
amino acids (methionine and cystine), the oil could be useful for cooking
purposes, and the high degree of unsaturation might enable it to be used
as a drying oil for paints and varnishes. It is a good source of minerals
required in human nutrition. The composition of the fatty acids and amino
acids and other aspects of the seed biochemistry have not been studied in
detail, although oil and protein contents of 29 and 30%, respectively,
have been reported by Asiegbu (1987).
2.4 Chemical composition of pigeon pea and pumpkin seeds:

Grain legumes provide food of high nutritive value to both humans and
their domestic animals. Quantitatively grain legumes are second to
cereals as a source of calories and protein El-Haradallou (1979). Most
dry pulses contain 20-25% protein which is about 2-3 times as much as
the protein content of cereal grains. The protein quality of legume grains
can be affected by factors such as processing and storage. The legume
grains contain anti-nutritional factors which can be destroyed by cooking
El-Hardallou (1979). There is very little data available in the literature
reviewed regarding the composition of the obtained pumpkin seeds as
well as the composition, chemical and physical characteristic of their oil
(Markovic and Bastic, 1976; El–Gharbawi, 1977; kamel, 1982).
2.4.1 Moisture Content:

Sanjeev et al., (1991) obtained a variation between 10.00 and 11.00%
moisture for six varieties of pigeon pea. El-Hardallou et al., (1980)
reported about 6.1% moisture content of legumes grown in Sudan. Papiti
(1970) reported about 5.0% moisture content of grain legumes grown in
Sudan. Ali (1996) obtained a variation between 4.50 and 6.25% moisture
content in four lines of pigeon pea grown in Sudan. The moisture content
6
of naked pumpkin seed (Cucurbita pepo) 6.03%, while, the seed cake of
naked pumpkin seed contained 8% moisture content El– Gharabawi
(1977,1978). Kamel (1982) reported 5.1% moisture content of pumpkin
from Canada (Cucurbita mixta). Ahmed (1985) found 3.90%, 4.50%
moisture content for white cultivar pumpkin seed and yellow cultivar
pumpkin seed flour 5.47%, while, Hamed (2006) obtained 6.10%
moisture content of processed pumpkin seed flour.
2.4.2 Crude protein content:

Singh and Eggum (1984) obtained a variation between 17.9 and 24.3%
crude protein of 43 commonly cultivated varieties of pigeon pea.
ICRISAT (1987) found a range in protein content from 18.2 to 19.80%
for six varieties of pigeon pea. Teixeira et al., (1985) found about 23.1%
protein content in pigeon pea cultivar kaki. Aletor and Aladetimi (1989)
obtained 21.7% protein content of cowpea varieties. About the same
value of 19.3% was observed by Al-Hardallou et al., (1980). Papiti
(1970) obtained 21.94%protein content of grain legumes grown in Sudan.
Ali (1996) obtained a variation between 21.24 and 23.23% crude protein
in four lines of pigeon pea grown in Sudan. Singh et al., (1990). Found
that the protein content of the high-protein genotypes of pigeon pea was
higher on average by nearly 20%. Pumpkin seed was considered as a
good source of protein as stated by Cirrilli (1971), who found 41.85%
protein content of Italy variety. El. Gharabawi (1977, 1978) obtained
29.94% protein of naked pumpkin seed (Cucurbita pepo), while, the seed
cake of naked pumpkin seed contained 55% protein. According to the
work carried out by Kim et al., (1978) the protein content of dehulled
seeds produced in Korea 25.5%. Kamel (1982) reported 34% protein
content of pumpkin from Canada (Cucurbita mixta). 59.8% protein of
pumpkin seed cake was reported by Zdunczyk et al., (1999), while, raw
7
defatted pumpkin seed without testa contained 69.7% protein Longe et
al., (1983). Yoon et al., (1983) reported that pumpkin and melon seeds
contain about 31% crude protein. Ahmed (1985) found 24.31% crude
protein for white cultivar pumpkin seed, and 34.13% crude protein for
yellow cultivar pumpkin seed. Hamed (2006) obtained 65.05%, 60.17%
protein for raw and processed pumpkin seed flour, respectively.
Sharama et al., (1986) reported that after oil extraction of pumpkin seed
flour the content of protein over 70% of dry mater.
2.4.3 Fat content:

Sanjeev et al., (1991) obtained a variation between 1.19 and 1.37% fat for
six varieties of pigeon pea. About the same value 2.86% was observed by
Aletor and Aladetimi (1989). While El-Hardallou et al., (1980) found
2.0% fat content of legumes grown in Sudan. Papiti (1970) found 1.75
fat content of grain legumes grown in Sudan. Ali (1996) obtained a
variation between 1.39 and 1.72% fat content in four lines of pigeon pea
grown in Sudan. Pumpkin seed was considered as a good source of oil as
stated by Cirrilli (1971) who found the oil content of Italy variety
45.35%. On the other hand, the oil content of naked pumpkin seed
(Cucrbita Pepo) was 38.65% El-Gharabawi (1977). According to the
work carried out by Kim et al., (1978) the oil content of dehulled seeds
produced in Korea were 46.5%. Kamel (1982) reported that the oil
content of seed kernel of pumpkin from Canada (Cucurbita mixta) on dry
basis was 41%, while, raw defatted pumpkin seed without testa contained
6.5% ether extract. Markovic and Bastic (1976) concluded that naked
pumpkin seeds contain 42–49% oil, while the normal seeds contain 30–
40% oil. Bhatia et al., (1977) reported a value of 40.6% total lipid for
pumpkin seed. Ahmed (1985) reported 41.56%, 36.28% oil content of
white and yellow cultivar pumpkin seed, respectively. Hamed (2006)
8
found 1.37% oil content of raw, and 1.43% oil content of processed
pumpkin seed flour.
2.4.4 Crude fiber content:

El Hardallou et al., (1980) found about 6.4% fiber content of legumes
grown in Sudan. About the same value 6.10% was observed by Aletor
and Aladetimi (1989), while, Ali (1996) obtained a variation between
8.59 and 11.63% crude fiber in four lines of pigeon pea grown in Sudan.
Cirrilli (1971) found 1.95% fiber content of Italy varity. The fiber
content of naked pumpkin seed (Cucurbita pepo) was 5.02%, while, the
seed cake of naked pumpkin seed contained 7.42% crud fiber EL-
Gharabawi (1977,1978). Kamed (1982) reported 3.74% fiber on seed
kernel of pumpkin from Canada (Cucurbita mixta). Zdunczyk et al.,
(1999) reported 3.17% crude fiber of pumpkin seed cake, while, raw
defatted pumpkin seed without testa contained 8.2% crude fiber Long et
al., (1983). Ahmed (1985) found 16.39% and 16.22% crud fiber content
for white and yellow cultivar pumpkin seed oil, respectively. Hamed
(2006) reported 2.98% fiber of raw pumpkin seed flour, 3.75% fiber of
processed pumpkin seed flour.
2.4.5 Ash content:

El-Hardallou et al., (1980) obtained 3.6% ash content of legumes grown
in Sudan. About the same value obtained by Aletor and Aladetimi
(1989). Papiti (1970) obtained 8.8% ash content of grain legumes grown
in Sudan. Ali (1996) obtained a variation between 4.67 and 4.95% ash
content in four lines of pigeon pea grown in Sudan. 4.7% ash content
was reported by Crilli (1971). On the other hand, the ash content of
naked pumpkin seed (Cucurbita pepo) was 5.47%, while the seed cake of
naked pumpkin seed contained 8.63% ash EL-Gharabawi (1977, 1978).
9
Kamel (1982) reported that the ash content of pumpkin from Canada
(Cucurbita mixta) was 4.13%. Zdunczyk et al., (1999) found 9.09% ash
content, while, raw defatted pumpkin seed without testa contained 9.3%
total ash Long et al., (1983). Ahmed (1985) reported 3.85% and 5.32%
ash content of white and yellow cultivar pumpkin seed oil, respectively.
While, Hamed (2006) found 9.04% ash content of raw sample and 8.78%
ash content of processed pumpkin seed flour.
2.4.6 Carbohydrate content:

Papiti (1970) obtained 55% carbohydrate. Sanjeev et al., (1991)
obtained a variation between 65.88 and 67.32% carbohydrate in six
varieties of pigeon pea, while El Hardallou et al., (1980) found 62.0%
carbohydrate. Ali (1996) obtained a variation between 54.64 and 58.21%
carbohydrate content in four lines of pigeon pea grown in Sudan. Kamel
(1982) reported 7.5% carbohydrate of seed kernel of pumpkin from
Canada (Cucurbita mixta). Hamed (2006) obtained 15.63% and 18.68%
carbohydrate of raw and processed pumpkin seed flour, respectively.
2.5 The nutritional value of seeds:

It is recognized that food grain legumes, such common bean, lentils and
kidney beans, represent the main supplementary protein source in cereal
and starchy food-based consumed by large sectors of the population
living in developing countries. Although, the nutritional value of these
legumes is of great importance, their intake is unfortunately lower than
that what is desirable. Furthermore, food grain legumes should be free of
antiphysiological substances, have high nutrient bioavailability and be
easily processed into edible, acceptable products (Bressani, 1989;
Bressiani, 1993). The nutritional value of grain legumes, not always fully
understood and accepted by consumers, is divided here into two large
10
groups: positive and negative factors. The positive factors include high
protein and lysine content, which allows legumes to serve as excellent
protein supplements to cereal grains (Bressani, 1989; Bressiani, 1993).
The health-related value of beans includes their positive effect on blood
cholesterol and glucose levels (Walker, 1982; Leeds, 1982), possibly
through the dietary fiber present in beans. The negative factors fall into
two groups. Antinutritional factors such as enzyme inhibitors, flatulence
factors, polyphenols, tannin and phytic acid. The negative nutritional
factors include protein and carbohydrate digestibility, sulfur amino acid
deficiency (Bressani, 1989; Bressiani, 1993).
2.6 Effect of processing on nutrients:

The heating process has four desirable effects ,first, it partially sterilizes
the food by killing microorganisms; second, it increases the availability
of nutrients by breaking down the plant cellulose cell walls that cannot be
broken down by the enzymes in the human intestinal tract Bradbury et al.,
(1984); third, it solubilises starch and makes it more digestible; fourth, it
denatures proteins, converting insoluble collagen in meat into soluble
gelatin and inactivating protolytic enzyme inhibitors that are potential
anti-nutritional factors present in many plant food Bradbury and
Holloway (1988). Several heat treatments (such as roasting) were
employed in pumpkin seeds preparations before they were consumed.
Cirrilli (1971) reported that pumpkin scoured seeds were employed for
human feeding after previous salting and roasting processes. The
apparent insignificant changes in the proximate composition (DM) with
the exception of ether extract and moisture content show the processed
were efficient in terms of nutrient retention Akinyele (1989). Giami et
al., (2001) reported that there were not significant differences on crude
protein, ash and fat content of raw and heat processed (roasted and
11
boiled) samples of breadfruit seed. Heat treatment for specific optimum
periods improved protein quality of cowpeas and other legumes by
improving protein digestibility, heating also inactivated anti-enzyme
factors (such as trypsin and amylase inhibitors), hemagglutinins,
polyphenols, and cyanogenic glycosides, which can influence protein
utilization. Heat treatment can also produce acceptable flavor and colors
with food legumes. However, extended cooking at high temperature and
pressures lowered nutritional quality of legumes Uzogara et al., (1992).
Clement et al., (1988) reported that cooking increases the IVPD from
71.8 to 83.5% in chickpea and also increases globulin and albumin
fractions. Giami and Wackuku (1997) reported that the processing
include roasting of some pulses consumed in Nigeria significantly
improved in vitro protein digestibility.
2.7 Antinutritional factors:

Anti-nutritional factors such as the haemagglutinin, saponins, tannin,
antivitamins, and phytic acid, which interferes with mineral, element
absorption and utilization and react with proteins to form complex
products which have inhibitory effect on peptic digestion. Of the various
antinutritional factors that are found in grain legumes, trypsin and
chymotrypsin inhibitors, amylase inhibitors, polyphenols (commonly
referred to as tannins), oligosaccharides, and phytic acid are important in
pigeon pea Singh (1988). In comparison with soy bean, pea, and
common bean, pigeon pea offers fewer antinutritional factor problems.
Pigeon pea contains considerably higher levels of protease inhibitors than
the other commonly consumed Indian grain legumes, but much lower
levels than those of soy bean Sumathi and patabhiraman (1975). Pigeon
pea contains considerable amounts of polyphenolic chymotrypsin, and
amylase. These are higher in pigeon pea cultivars with dark seed-coat
12
colour Singh (1984). Phytolectins are toxic factors that interact with
glycoprotein on the surface of phytolectins which are highly sensitive to
heat treatment and hence may be of little nutritional significance. Pigeon
pea contains traces of glycosides but not at toxic levels (Singh, 1988).
Researches (Bressani et al., 1984; Bressani, 1985; Bressani, 1989;
Bressani, 1993; Bressani and Sosa, 1990) found that traditional recipes,
such as cooking, soaking, dehulling and germination help not only in
making beans more acceptable to consumers but also significantly
reducing any antinutritional elements in the food grain legumes. The most
common way to process food legumes is to subject them to thermal
processing. Although the main goal of thermal processing is to render the
grain soft, its effects go beyond the changes in physical structure and
texture (Bressani et al., 1984; Bressani, 1985; Bressani, 1989; Bressani,
1993; Bressani and Sosa, 1990). Although heat treatment will effectively
eliminate most of these undesirable substances, the application of other
process such as soaking, cooking, steeping, decorticating, or germination
have also been effective in reducing anti-nutritional factors Fadul (1998).
2.7.1 Phytic acid of pigeon pea and pumpkin seeds:

Early studies showed that foods rich in phytate or sodium phytate added
extrinsically to diets were the main causes of decreased overall calcium,
iron, and zinc balance in rats and human (Reinhold et al., 1976; Oberleas,
1983). Phytates have been found in cereal grains and legumes up to a
level of approximately 5% by weight (de Boland et al., 1975). Although
phytase is present in mature seeds, it appears to have little effect on
phytate in dry or dormant seeds. Generally, phytic acid is thought to be
abundant in the outer layer of monocotolydones seeds. While high levels
of phytic acid were found in globoids of dicotyledonous seeds. In cereals
grains, it is distributed on both the bran and germ, except for corn in the
13
germ. The amount of phytic acid varies from 0.5 to 6% in cereal,
legumes and oil seeds accounts from 60 to 90% of their total phosphorus
content (Fox and Toa, 1989). In pearl millet phytic acid content
represents more than 70% of the total phosphrus in the grain Chauhan et
al., (1986). Mahajan and Chauhan (1990) found that pearl millet contains
835 mg/100g. A value of 990mg/100g was reported by Khetarpaul and
Chauhan (1989), while Kumar and Chauhan (1993) gave a value of 825.7
mg/100g. The molecule inositol hexaphosphate and salt ions of it are
commonly referred to as phytat. These molecules tend to form insoluble
complexes with protein and calcium and /or zinc ions, which make them
less available for absorption and utilization Petterson (2000). Giami et al.,
(2004) reported that the level of phytic acid in raw seed (1.19mg/g) was
lower than the levels found in some commonly consumed pulses in
Nigeria. Phytic acid content of pumpkin seed kernel flour was 2.37
g/100g as reported by El-Adawy and Taha (2001).
2.7.2 Tannins content of pigeon pea and pumpkin seeds:

Tannins are the group of polyphenolic compounds that bind to proteins
and either inhibit their activity in the case of digestive enzymes or
complex with proteins. Tannins can also form cross linkages between
protein and other macromolecules and render them unavailable for
digestion Griffith (1991). Furthermore tannins can bind other endogenous
proteins in the intestinal tract such as digestive enzymes (lipase,
protolytic enzyme, amylases) and inhibit them. Van Wyle et al. (1973)
reported that in general, tannin content appeared in inverse relation to
protein digestibility the relation was by no means constant or uniform, for
example, where the tannin content appeared as 0.95%, digestibility
ranged from 24 to 62%. Considerably, in legumes higher levels of
tannins are generally found in some varieties of soybeans, field peas and
14
faba beans Petterson (2000). ICRISAT (1981) obtained a variation
between 0.003 and 0.015 % tannins of pigeon pea. While, Singh and
Eggum (1984) found a variation between 0 and 0.2 % tannins. El-Adawy
and Taha (2001) found that the tannin content of pumpkin seed kernel
was 0.17 g/100g.
2.7.3 Polyphenols of pigeon pea and pumpkin seeds:

Polyphenols constitute one of the most numerous and ubiquitous groups
of plant metabolites and are integral part of human and animal diet.
Ranging from simple phenolic molecules to highly polymerized
compounds. The occurrence of this complex group of substances in plant
foods is extremely variable. Polyphenolis traditionally have been
considered anti-nutrient by nutritionists, because of the adverse effect of
tannins, one type of polyphenolis, on protein digestibility. However,
recent interest in food phenolics has increased greatly, owing to their
antioxidant capacity (free radical scavenging and metal chelating
activities) and their possible beneficial implications in human health
Bravo (1998). Polyphenolic content in cereals usually less than 1% of
dry matter except for some sorghum cultivars which can have as much as
10 % (Bravo, 1998). Glycosylvitexin, glucosylorientin and viterin are
the phenolic compounds concentrated in the bran of pearl millet and
influences the colour of the millet according to acidic or alkaline pH
Nugdallah (2003). These compounds as suggested by Birzer and
Klopfenstein (1988) is the goitrogen in pearl millet in addition to the
thioamide and other compounds derived from flavonoids.
2.8 Effect of processing on antinutritional factors:

Cooking of seeds has a little effect on phytate levels of cereals and
legumes and about 17 to 31 % reduction was in cereals and 16.8 to 17%
15
in legumes, Maga (1982). Lyer et al., (1980) observed that soaking of
beans such as Phaseolus vulgaris followed by cooking significantly
reduced phytic acid. Alonos et al., (2000) reported that dehulling of faba
and kidney beans increased the level of phytic acid. Carnovale et al.,
(1988) showed that dehulling concentrated the protein and phytic acid
content. Dehulling of beans significantly elevated the percentage of
phytic acid compared the whole beans Deshpande et al., (1982). Zia-ur-
Rehman (2005) found phytic acid content in food legumes ranged from
970 to 1440 mg/100g, and it reduced by 28.0-516%, as a result of
cooking. Bressani and Elias (1980) observaed that about 30 to 40 % of
tannin could be removed from Phaseolus vulgaris by cooking. Moreover
Bressani et al., (1991) found that cooking of Phaseolus vulgaris reduced
tannic acid by 6.2%. Since tannins are mainly located in the testa, its
physical removal (dehulling) significantly decrease tannin concentration
as reported by Alonso et al., (2000). The infrared, autoclaving and
boiling-water treatments destroyed most of the trypsin inhibitor and
haemagglutinating activities, and reduced the level of tannins Kadam et
al., (1987). Tannins content in five food legumes ranged from 770 to
1100mg/100g, and it was reduced by 33.1-45.7% after cooking Zia-ur-
Rehman (2005).
2.9 In vitro protein digestibility (IVPD) of pigeon pea and pumpkin

seeds:
The protein quality of food or feed depend on its amino acid composition
and digestibility. The protein digestibility primarily determines the
availability of its amino acids Hahn et al., (1981). One of the main
factors affecting the nutritive value of legume proteins is their limited
susceptibility to hydrolysis by digestive enzymes. This proteolyic
resistance has been attributed to the structural characteristic as well as to
16
the presence of anti-nutritional compounds such as trypsin inhibitors,
polyphenols, and phytic acid Clemente et al., (2000). Kumar and Singh
(1984) reported that the condensed tannin present in sorghum might have
inhibited the activities of proteolytic enzymes like pepsin digestibility in
high concentration of tannin. Low digestibility is a major nutritional
problem in high-tannin sorghum cultivars. The digestive of protein
appears to depend on phytic acid in the diet. Carnovale et al., (1988)
reported that a reduction of 6.8, 5.7 and 8.7%, respectively, in
digestibility of whole flour, protein concentrates, and protein isolates of
faba bean samples following the addition of 10mg of phytic acid.
Sangronis and Machado (2005) found 76.8% in vitro protein digestibility
in Phaseolus vulgaris. Singh and Euggm (1984) found a variation
between 57.9 and 64.1% in vitro protein digestibility of pigeon pea, while
Sanjeev et al., (1991) obtained a variation between 32.10 and 38.10%
protein digestibility in six varieties of pigeon pea. Ali (1996) obtained a
variation between 62.40 and 69.64% protein digestibility in four lines of
pigeon pea growth in Sudan. The protein digestibility of the raw food
legumes ranged from 33.8 to 37.6%. In vitro protein digestibility of
pumpkin seed Kernel flour was 90% and for watermelon seed kernel
flour was 87.9% reported by El-Adawy and Taha (2001).
2.10 Effect of processing on In vitro protein digestibility (IVPD):

Processing can improve the digestibility of legume protein by the
destruction of trypsin inhibitor and the opening of protein structure
through denaturation El-faki et al. (1984). Many researchers (Bressani
et al., 1982; Bressani, 1984; Bressani et al., 1988; Bressani, 1993)
observed that dehulling followd by cooking increased protein quality and
digestibility. Dehulling was found to improve protein quality and protein
digestibility due to the removal the seed coat tannin, Berssani et al.,
17
(1984), which may contribute to decrease protein digestibility. True
protein digestibility was significantly increased by cooking in both
whole–seed and dhal samples Singh et al., (1990). However, cooking in
boiling water resulted in improvement in protein of the food legumes by
86.9 and 93.3% Zia-ur-Rehman (2005). Cooking significantly increased
protein digestibility for the control and all treated samples. Elsheikh et
al., (2000). Giami and wachuku (1996) reported that the processing
(roasting, boiling, dehulling and soaking) significantly (p ≤ 0.01)
improved in vitro protein digestibility. Removal of tannin through
chemical or physical treatment was found to improve in vitro protein
digestibility and weight gain, but removal or lowering the content of
tannin through genetic means is an important goal in both cereal and
legumes FAO (1981).
2.11 Protein quality of seeds:

The protein quality of pigeon pea is primarily expressed in terms of its
protein content, the levels of amino acids and protein digestibility Singh
and Eggum (1984). In most food crops, genetic variability for protein
content is considered an important factor for improving protein quality by
selection and breeding. However, environment and agronomic practices
influence the protein quality of pigeon pea to a considerable extent, and
this should be noted while breeding for protein quality. The sulphur
amino acids, methionine and cysteine, are the most limiting amino acids
of legumes, and very low values for these amino acids were reported in
pigeon pea Eggum and Beames (1983). In legumes negative relationships
are usually found between protein percentage and methionine content per
unit of protein Bliss and Hall (1977). However, there was no strong
relationship between methionine (g/100g protein) and protein (%) in
pigeon pea, indicating that both protein and methionine could be
18
improved Singh and Eggum (1984). Seed protein fractions play an
important role in determining the over all amino acid composition of seed
proteins. Further, the globulin proteins are deficient in sulphur amino
acids. Although present in a small proportion, albumin fractions, are a
very rich source of methionine and cystein. Glutelin fraction is also a
better source of sulphur amino acids than globulin, and hence may be
nutritionally desirable.
2.12 Protein fractions of pigeon pea and pumpkin seeds:

Fractions (1,2) include water soluble and salt-soluble proteins which
constitute over 84% of total protein, extracted with distilled water and
sodium chloride respectively. Fractions (3,4) include the ethanol-soluble
protein fractions and the alkali-soluble protein fractions which constitute
0.97% and 11.0%, respectively, (Bollini and chrispeels,1978; Youle and
Huang,1978; Deshpande and Nielsen,1987).
2.12.1 Albumins and globulins:

Albumins and globulins are very heterogeneous and differed in
polypeptide composition. Extraction with water and then with NaCl to
solubilize what is termed albumins and globulins respectively. The
globulin constituted 82.6% of the total seed proteins and the albumins
were richest (3.76 g/100g protein) in methionine, Dhankher et al., (1990).
Water-and salt-soluble proteins together accounted for 63 to 83%of the
total nitrogen of chickpeas, cowpeas and dry beans Deshpande et al.,
(1987). The general deficiency of lysine in most cereals e.g sorghum and
corn is essentially the consequence of their low content of albumins and
globulins FAO (1981). In terms of composition the albumins are richer
in the sulpur amino acids and other essential amino acids than the
globulins in pigeon pea Singh and Jambunathan (1982). Globulins are
19
storage proteins used during germination, and form discrete bodies bound
to the cell membranes. They are most deficient in sulphur– amino acids
in pigeon pea Singh and Jambunathan (1982). Nugdallah and El Tinay
(1997) reported that albumins and globulins ranged from 4.0 to 12% and
66 to 80% respectively. Singh et al., (1981) found 70.4% albumin and
globulin together in decorticated seed of pigeon pea. Ali (1996) obtained
a variation between 63.54 and 66.41% Albumin fraction, 5.48 and 6.38%
globulin fractions in four lines of pigeon pea growth in Sudan. Albumin
and globulin fractions were found to be the major seed proteins of fluted
pumpkin seeds, constituting about 58.6% of the total protein of
ungerminated (raw) seeds, also, albumin and globulin protein fractions
were found to be the major seed proteins of African bread fruit seeds,
constituting about 67.8% of the total protein of the raw seed Giami et al.,
(2004).
2.12.2 Prolamin:
Prolamin can be defined as a portion material extracted at room
temperature by aqueous alcohol; free of reductant and salt, from a corn
meal deprived of lipids and salt soluble protein. Prolamin content ranged
from 3.1 to 6.9% of total seed protein in chickpea seeds Dhawan et al.,
(1991). Prolamin content was 2.6% in cowpea seeds Dhankher et al.,
(1990). The ethanol soluble protein fraction was 0.97% in cowpea seeds
Deshpande and Nielsen (1987). Nugdallah and El Tinay (1997) reported
that prolamins ranged from 1.4 and 4.0% for cowpea. Singh and
Jambunathan (1982) found 3.00% prolamin in whole seed of pigeon pea.
While, Singh et al., (1981) obtained 3.1% prolamin fractions in
decorticated seed of pigeon pea. Ali (1996) obtained a variation between
1.95 and 2.68% prolamin fractions in four lines of pigeon pea growth in
20
Sudan. The prolamin fraction was observed to decreased by 54.6% as
a result of germination.
2.12.3 Glutelin:
Glutelin contains higher levels of suphur–amino acids than globulins in
pigeon pea Singh and Jambunathan (1982). Singh et al., (1981) reported
17.4% glutelin in whole seed of pigeon pea. About 78% of the pigeon
pea seed proteins were salt soluble, out of which 61% were globulins
which were further separated into three fractions Gopalakrishna et al.
(1977). Singh and Jambunathan (1982) obtained about 70.1% water–and
salt–soluble protein fractions together in whole seeds of pigeon pea. Ali
(1996) obtained a variation between 10.75 and 14.25% glutelin fractions
in four lines of pigeon pea growth in Sudan. While, Singh et al., (1981)
obtained 19.6% glutelin fractions in decorticated seed of pigeon pea. The
glutelin fraction was observed to increase by 57.0% as a result of
germination.
2.13 Effect of processing on protein fractions:

The effect of cooking on protein quality, in terms of amino acids and
bioavailability of legume proteins, is important. A slight reduction in
lysine content was observed as a result of cooking Singh et al., (1990).
Protein quality of pigeon pea has been reported to be greatly influenced
by storage and processing practices Singh and Eggum (1984).
2.14 Digestibility of protein major fractions:

Legumes are major crops in many tropical countries and serve as
important sources of carbohydrates and proteins to poor populations of
these regions Singh and Rachie (1985). Most of the literature on the
degradation of legumes proteins concerns the major storage protein
21
(globulins), which constitutes 50 to 70% of total seed proteins (Khan et
al., 1980; Shewry, 1995). This protein fraction is free of antinutritional
factors, such as proteinase inhibitors and lectins, and this is why it is
often used in studies designed to answer questions related to the
nutritional value of proteins. Most studies designed to answer question
related to nutritional quality of legume proteins have focused on proteins
in the globulin fractions are present in the seeds in high amounts and is in
fractions of antinutritional factors Shewry (1995). Bean (Phaseolus
vulgaris) proteins have a low nutrional value due to reduced digestibility
and low methionine content and bioavailability Bressani (1993).
phaseolin (or G1 globulin) is the major seed storage protein and
represent, along with albumins, near 80% of total bean proteins.
Phaseeolin is very poorly digested when in its native state, but effectively
hydrolysed after heat treatment (Liener and Thopson, 1980; Deshpande
and Nielsen, 1987). The in vitro digestibility of albumin proteins is low
(26-32%) and after heating it is reduced even more, to 13-18%,
independently of bean variety (Marquez and Lajolo, 1981; Genoveses and
Lajolo, 1996). Methionine content is similar for the two fractions;
although the in vitro bioavailability, after heat treatment, is high for
phaseolin but very low for albumins, as expected from digestibility values
Genovese and Lajolo (1996). On the other hand, Coelho and Sgarbieri
(1995), working with albumins separated into acid- and alkali-extracted
fractions, did not observed any reductions of digestibility of provoked by
heating, and a degree of in vitro hydrolysis similar to that of phaseolin
was attained. The in vitro digestibility of bean protein fraction was low
when in the native state and was differently affected by denaturation. For
phaseolin, the main reserve protein, heating caused a significant increase
of susceptibility to hydrolysis, whereas heat had no apparent effect on
digestibility of glutelins and albumins. For the PIL (protease inhibitor-
22
lectin rich) fraction, which was shown to have a composition similar to
total albumins, there was a decrease of digestibility, probably associated
to disulfide bond formation upon heating Genovese and Lajolo (1998).
Anibaba and Osagie (1997) found a continuous degradation of prolamin
and glutelin (storage proteins) with a concomitant rise in albumin and
globulin (enzyme proteins) resulting in a synchronous rise in free amino
nitrogen (FAN) during malting of sorghum. On the other hand, all the
major proteins in farafara increased at the peak of malting without a
synchronous increase in FAN. Genovese and Lajolo (1998) obtained 28.8
and 19.4% for native and heated albumins after pepsin and pancreatin
digestion, respectively, also obtained 72.9 and 45.3% of 5% TCA soluble
albumins, after pepsin-pancreatin digestion, respectively, of Phaseolus
vulgaris.
23
CHAPTER THREE
MATERIAL AND METHOD
3-1 Materials:
A cultivar of pigeon pea seeds (Cajanus cajan ) was obtained from the
Department of the Agricultural Research Corporation, Madani, also seeds
of pumpkin cultivar was obtained from local market, Khartoum North,
were employed for all determinations. All chemicals used in this study
were of reagent grade.
3-2 Methods:-
3.2.1 Preparation of samples:
The pigeon pea seeds were cleaned and freed from foreign material.
Then divided into two groups one group used as raw, the other group was
cooked in distilled water for 1/2 hr, the water decanted and the seeds
dried, then each one divided into two groups one milled with coat and the
other dehulled then milled. The Pumpkin seeds were cleaned and freed
from foreign materials. The seeds were divided into two groups. One
group was roasted and the other used as a raw. The seed hulls were
removed manually milled in electric miller and then defatted and dried.
3.2.2 Defatting of pigeon pea and pumpkin seeds:

Cold extraction method was used to extract oil. In this method, each of
seeds flour was placed in a conical flask and mixed with hexane (1:10),
the mixture was shaken for 16 hours in a mechanical shaker and then
filtered. The filtrate was washed again with another volume of hexane to
remove traces of the oil. The mixture was then centrifuged to separate
the solvent. The ether extract free flour was dried in open air at room
temperature. The dried flour was then ground to pass through 0.4 mm
mesh and stored at 0ºC for further analysis.
24
3.2.3 Analytical methods:
3.2.3.1 Proximate analysis of samples:
The determination of moisture, crude fiber, crude protein, fat and ash
contents were carried out on the both raw and processed samples flour
according to the A.O.A.C (1984) methods.
3.2.3.2. Determination of anti-nutritional factors:

3.2.3.2.1. Determination of tannin content:
Quantitative estimation of tannin for each sample was carried out using
modified vanillin-HCl in methanol method as described by Price et al.,
(1978). About 0.2g of the ground sample was placed in a 100 ml conical
flask. Ten milliliters of 1% HCl in methanol (v/v) were added, shaken for
20 min, and centrifuged at 2500 rpm for 5 min. One milliliter of the
supernatant was pippetted into a test tube and 5 ml of vanillin–HCl
reagent were added. The optical density was read using
Spectrophotometer (JENWAY 6305 UV/V) at 500 nm after 20 minutes
incubation at 30ºC. A standard curve was prepared expressing the results
as catechin equivalent, i.e. amount of catechin (mg per ml) which gives a
colour intensity equivalent to that given by tannins after correcting for
blank.
Calculations:
Tannin concentration was expressed as catechin equivalent (C.E) as
follows:
C × 10 × 100
C.E % =
200
25
Where:
C = Concentration corresponding to the optical density.
10= volume of extract in ml.
200= sample weight in mg.
3.2.3.2.2. Determination of phytic acid:

Phytic acid content was determined according to the method described by
Wheeler and Ferrel (1971). Two grams of dried sample were weighed in
125 ml conical flask. The sample was extracted with 50 ml of 3%
tricholoro acetic acid (TCA) for 3hr with mechanical shaking. The
supernatant was centrifuged at 3000 rpm for 5 min. 10 ml aliquot of
supernatant was transferred to a 40 ml tube and 4 ml of FeClз (FeClз
+3
solution containing 2mg F ion/ml 3% TCA) were then added to aliquot.
The tube was heated in a boiling water bath for 45min. One or two drops
of 3% sodium sulfate dissolved in 3% TCA were added. The tube was
cooled and centrifuged for 10-15 min and the clear supernatant was
decanted. The precipitate was washed twice by dispersing well in 25ml
3% TCA, heated for 10-15 min. In a boiling water bath and then
centrifuged again. Washing was repeated once more with distilled water,
the washed precipitate was dispersed in few milliliters of distilled water
enriched with 3ml of 1.5 N NaOH, and the volume completed to
approximately 30ml with distilled water. Heated in boiling water bath for
30min. and hot filtered using Whatman NO.2. The precipitate was
washed with 60-70 ml hot water, and the washings were decanted, the
precipitate from the filter paper was dissolved in 40ml hot 3.2 N HNOз
and placed in a 100 ml volumetric flask. The paper was washed with hot
distilled water and the washing was collected in the same flask then
completed to volume. A 0.5 ml of aliquot was taken from the above
solution and transferred into 10 ml volumetric flask. Then 2 ml of 1.5 N
26
KSCN (potassium thiocyanate) were added and completed to volume by
distilled water then immediately (within one min.) read at 480nm using
(JENWAY 6305 UV/V is spectrophotometer). A standard curve of
different Fe (NOз) з concentrations was plotted to calculate the ferric ion
concentration. The phytate phosphorus was calculated from the iron
concentration assuming 4:6 irons to phosphorus molar ratio.
3.2.3.2.3 Determination of polyphenols:

polyghenols content was determined according to the method described
by Price and Butler (1977). Ground sample (60mg) was extracted with 3
ml absolute methanol in a test tube, by constant shaking for one minute,
then poured into a filter paper. The tube was quickly rinsed with an
additional 3 ml of methanol and the contents poured at once into the filter
paper. The filtrate was diluted to 50 ml with distilled water, mixed with 3
ml 0.1 M Fe Cl3 in 0.1 N Hcl for 3 minutes, followed by the timed
addition of 3 ml 0.008 M K3 Fe (CN)6. The absorption was read after 10
minutes at 720 nm on aspectrophotometer (corning, 259). In all cases,
tannic acid was used as a reference standared.
3.2.3.3 In vitro protein digestibility determination:

In vitro protein digestibility of raw and processed sample was measured
according to the method of Saunders et al. (1973). Exact 250 mg seed
flour was suspended in ml of 0.1 N .HCL containing 1.5 mg pepsin
(1:10,000) in a 100 ml conical flask. The mixture was incubated at 37 ºC
for 3 hours only. The mixture was then neutralized with 0.5 N NaOH and
treated with 4 mg panereatin (Grade VI porcine ) in 7.5 ml of 0.2 M
phosphate buffer (PH 8.0), containing 0.005 M sodium azide. The
mixture solution was incubated at 37 ºC for 24 hours. Ten milliliters of
10% trichloro acetic acid (TCA) were added to the mixture to stop the
27
reaction the mixture was then centrifuged at 5000 rpm for 5minutes. Five
milliliters aliquots from the supernatant were pipetted analyzed for
nitrogen content.
Calculation:
Protein digestibility % = N in supernatant – enzyme N x 100
N in sample
3.2.4 Physiochemical properties:

3.2.4.1 Protein fractionation due to solubility:
The Mendel–Osborne (1914) technique for protein fractionation was used
in this study.
3.2.4.1.1 Determination of water–soluble proteins:

A sample of 2 grams was taken from defatted seed flour for fractionation
of total protein. To this amount of the flours, 3 volumes of 30 ml distilled
water was added and the mixture was shaken for 30 minutes using a
mechanical shaker, then centrifuged at 3000 rpm for 20 minutes to
separate the insoluble part from the liquor. About 10ml of the solvent
were taken for protein estimation according to the micro-kjeldahl method.
The following formula was used for calculating percentage of albumin:
Total albumin = T x 14x D.F x 6.25 x N

Wt x 1000
Where:
N: Normality of acid (0.02)
T: Titer reading.
D.F: Dilution factor.
28
Wt: Weight of sample.
14: Each ml HCl is equivalent to 14 mg nitrogen.
Albumin % = Total albumin x 100

Total protein
3.2.4.1.2 Determination of salt soluble proteins:

The insoluble part obtained after extraction of albumin was re extracted
with 3 volumes of 30 ml NaCL (I M) for 30 minutes with continuous
shaking. The mixture was then centrifuged at 3000 rpm for 20 minutes to
separate insoluble part. The extracted liquor was collected in containers.
About 10 of the liquor were taken for estimation of soluble protein by the
micro-kjeldahl method:
Total Globulin = T x 14x D.F x 6.25 x N

Wt x 1000
Globulin % = Total globulin x 100

Total protein
3.2.4.1.3 Determination of alcohol soluble proteins:

The insoluble part obtained after the extraction of prolamin was re-
extracted with 3 volumes of 30 ml 70 % ethanol to the insoluble part with
continuous shaking for 30 minutes in a mechanical shaker. The peptized
liquor was collected in containers. About 10 ml of the liquor were taken
for protein determination by the micro-kjeldahl method. The percentage
of alcohol soluble proteins was calculated from total proteins as follows:
Total Prolamin = T x 14x D.F x 6.25 x N
Wt x 1000
29
Prolamin % = Total prolamin x 100
Total protein
3.2.4.1.4 Determination of alkali-soluble proteins:

The insoluble part obtained after the extraction of prolamin was re-
extracted with 3 volumes of 30 ml NaOH (0.2%) for 30 minutes with
continuous shaking. The insoluble part was separated by centrifugation
at 3000 rpm for 20 minutes. The peptized liquor was collected and 10 ml
taken for nitrogen determination
Total Glutelin = T x 14 x D.F x 6.25 x N

Wt x 1000
Glutelin % = Total glutelin x 100
Total protein
3.2.4.1.5 Protein content of insoluble fraction:

The remaining insoluble part of the sample was dried and 0.2 grams was
digested in 3.5 ml sulfuric acid and used for estimation of insoluble
nitrogen.
3.2.5 Statistical analysis:

Data generated was subjected to Statistical Package for Social Sciences
(SPSS). Means (±SD) were tested using one factor analysis of variance
(ANOVA), and then separated using Duncans Multiple Range Test
(DMRT) according to Mead and Gurnow (1983).
30
CHAPTER FOUR
RESULT AND DISCUSSTION
4.1 Effect of processing on the proximate composition of defatted

pigeon pea seeds:
The proximate composition of both treated and untreated samples are
illustrated in Table 1. The moisture content of untreated sample was
found to be 10.80%, treatments used had no significant (p ≤ 0.05) effect
on moisture content which reduced to 9.08% after dehulling, 9.00% after
cooking and 8.90% after cooking followed by dehulling. The protein
content of defatted untreated sample was found to be 22.57%, treatments
used had no significant (p ≤ 0.05) effect on protein content which
increased to 23.41% as a result of dehulling, and decreased to 20.12%
and 21.94% as a result of cooking, cooking followed by dehulling,
respectively. The increase of protein content after dehulling may by due
to removal of seed coat. Ash content of untreated sample was found to be
4.65%, dehulling and cooking had no significant (p ≤ 0.05) effect on ash
content which were reduced it to 4.14% and 3.91%, respectively.
However, cooking followed by dehulling significantly (p ≤ 0.05)
decreased ash content to 2.91%. Oil content of untreated sample was
found to be 2.78%. Treatments applied had no significant (p ≤ 0.05)
effect on oil content which were reduced it to 2.60% after dehulling,
2.03% after cooking and 1.75% after cooking followed by dehulling.
Fiber content of untreated sample was found to be 10.11%. Treatments
applied had no significant (p ≤ 0.05) effect on fiber content which was
reduced to 9.12% after dehulling, to 8.86% after cooking and to 8.30%
after cooking followed by dehulling. Reduction in protein, ash, oil and
fiber content after cooking and cooking followed by dehulling may be
due to leaching in water. Carbohydrate content of untreated sample was
31
found to be 46.60%, dehulling and cooking had no significant (p ≤ 0.05)
effect on carbohydrate content, while, cooking followed by dehulling
significantly (p ≤ 0.05) increased carbohydrate to 58.21%.
4.2 Effect of roasting on the proximate composition of defatted

pumpkin seeds:
The proximate composition of both treated and untreated samples are
illustrated in Table 1. The moisture content of untreated sample was
found to be 5.71%, roasting of pumpkin seed significantly (p ≤ 0.05)
decreased moisture content to 4.50 %. The protein content of untreated
sample was found to be 77.79%, roasting significantly (p ≤ 0.05) reduced
the protein content to 73.02%. The value for protein obtained in this
study was higher than the values obtained for the full fat pumpkin seeds
34% and 25.5% reported by Kamel (1982) and Kim et al., (1978),
respectively, which possibly attributed to defatting process. Moreover,
the result agreed with Sharama et al., (1986) who reported that defatting
of pumpkin seed flour increased the content of the protein over 70% on
dry weight base. The reduction in protein content after roasting may be
due to the effect of defatting and may be due to coagulation of protein.
Bradbury and Halloway (1988) reported that heating process denatures
the protein. The ash content of untreated sample was found to be 8.30%,
roasting of pumpkin seed significantly (p ≤ 0.05) decreased ash content to
6.07%. Oil content of untreated sample was found to be 1.54%, roasting
of pumpkin seed significantly (p ≤ 0.05) increased oil content to 2.78%.
Fiber content of untreated sample was found to be 2.61%, roasting of
pumpkin seed significantly (p ≤ 0.05) increased fiber content to 4.05%.
Carbohydrate content of untreated sample was found to be 5.28%,
32
Table 1: The proximate composition (%) of defatted pigeon pea and pumpkin seeds
Treatments Moisture Crude Crude ash Oil Crude fiber Carbohydrate

protein
(A) Pigeon pea seeds:
Untreated 10.80(±3.11)a 22.57(±3.59)a 4.65(±0.16)a 2.78(±2.72)a 10.11(±2.15)a 46.60(±7.77)b
Dehulled 9.08(±3.11)a 23.41(±0.30)a 4.14(±0.40)ab 2.60(±0.35)a 9.12(±0.15)a 51.67(±0.00)ab
Cooked 9.00(±0.00)a 20.12(±1.87)a 3.91(±0.80)ab 2.03(±0.04)a 8.86(±0.09)a 53.60(±1.53)ab
Cooked dehulled 8.90(±0.49)a 21.94(±0.28)a 2.91(±0.84)b 1.75(±1.41)a 8.30(±0.35)a 58.21(±0.98)a
(B) Pumpkin seeds:
Unroasted 5.71(±0.43)a 77.79(±1.75)a 8.30(±0.36)a 1.54(±0.64)b 2.61(±0.53)b 5.28(±3.99)b
Roasted 4 .50(±0.71)b 73.02(±2.95)b 6.07(±0.21)b 2.78(±0.31)a 4.05(±0.42)a 8.39(±3.70)a
Values are means (±SD). * Mean values having different superscript letters in columns differ significantly (P ≤ 0.05).
33
roasting of the seed significantly (p ≤ 0.05) increased carbohydrate
content to 8.39%.
4.3 Effect of processing on antinutritional factors of defatted pigeon

pea seeds:
The antinutritional factors (phytic acid, tannins and polyphenols) of both
treated and untreated pigeon pea seeds are illustreated in Table 2. Phytic
acid content of untreated sample was found to be 158.44mg/100g,
dehulling of the seeds significantly (p ≤ 0.05) affect phytic acid content
and increased it to 173.16mg/100g. However, cooking was significantly
(p ≤ 0.05) reduced the phytic acid content to 132.96mg/100g, while,
cooking followed by dehulling reduce it to 136.03mg/100g. The
increment in phytic acid content due to removing the coat. Tannins
content of untreated sample was found to be 238 mg/100g. Treatments
applied reduced tannin content. Dehulling was found to reduce it to 205
mg/100g, while, cooking reduce it to 150 mg/100g, cooking followed by
dehulling significantly (p ≤ 0.05) affect tannin content and reduced it to
70 mg/100g. Polyphenols content of untreated sample was found to be
256.005mg/100g. All treatments applied were significantly (p ≤ 0.05)
reduced polyphenols content and was found to be 107.625 after dehulling,
to 74.750 after cooking and to 61.235mg/100g after cooking followed by
dehulling. The effect of heat treatments on the antinutrational factors
agreed with Hassan (2004), Nugdallah (2003), Zia-ur-Rehman (2005) and
others. Fadul (1998) reported that the application of processes such as
cooking have been effective in reducing the antinutrational factors.
34
4.4 Effect of roasting on antinutritional factors of defatted pumpkin
seeds:
The antinutritional factors (tannin, phytic acid and polyphenols) of both
treated and untreated pumpkin seeds are illustrated in Table 2. Phytic
acid content of untreated sample was found to be 216.07mg/100g,
roasting had no significant (p ≤ 0.05) effect on phytic acid content which
reduced it to 208.41mg/100g. Tannin content of untreated sample was
found to be 698 mg/100g, roasting significantly (p ≤ 0.05) reduced
tannins content to 142 mg/100g. Polyphenols content of untreated
sample was found to be 77.455mg/100g, roasting significantly (p ≤ 0.05)
reduced polyphenols content to 55.840mg/100g. The reduction in tannin,
polyphenols and phytic acid content of roasted sample possibly result
from the effect of the heat treatment. Fadul (1998) reported that the
application of processes such as cooking have been effective in reducing
the antinutritional factors.
4.5 Effect of processing on in vitro protein digestibility (IVPD) of

defatted pigeon pea seeds:
The in vitro protein digestibility of both treated and untreated pigeon pea
seeds are illustreated in Figure 1. The IVPD of untreated sample was
found to be 61.25%. Treatments applied increased the IVPD to 72.56%
after dehulling, to 84.40% after cooking and to 91.25% after cooking
followed by dehulling. The increment in IVPD after dehulling may be
attributed to the removal of tannin and fiber which are mainly located in
the seed coat (Alonso et al., 2000; Bressani et al., 1984). The increment
in IVPD for the cooked sample may be due to reduction of antinutritional
factors as a result of cooking. This observation was agreed with that
reported by Bressani et al., (1984), who found that dehulling, dehulling
followed by cooking increased protein quality and digestibility.
35
Table 2: Antinutritional factors (mg/100g) of defatted pigeon pea
and pumpkin seeds
Treatments Phytic acid Tannins Polyphenols
Untreated 158.44(±6.23)b 238(±0.05)a 256.005(±15.61)a
Dehulled 173.16(±1.60)a 205(±0.01)a 107.625(±3.19)b
Cooked 132.96(±2.87)c 150(±0.00)ab 74.750(±1.27)c
Cooked dehulled 136.03(±4.36)c 70(±0.03)b 61.235(±1.28)c
(B) Pumpkin seeds:
Unroasted 216.07(±0.00)a 698(±0.00)a 77.455(±3.19)a
Roasted 208.41(±7.18)a 142(±0.01)b 55.840(±0.00)b
* Values are means (±SD).

* Mean values having different superscript letters in columns differ
significantly (P ≤ 0.05).
36
4.4 Effect of roasting on in vitro protein digestibility (IVPD) of
defatted pumpkin seeds:
The in vitro protein digestibility of both treated and untreated pumpkin
seeds are illustreated in Figure 2. The IVPD of untreated sample was
found to be 42.80%, roasting process improved the IVPD and was found
to be 64.38%. The results obtained after roasting was agreed with Hamed
(2006), Hassan (2004) and also agreed with those reported by Giami and
Wackuku (1996) who found that the processing (include roasting) of
some pulses significantly improved the IVPD. The increment in IVPD
of roasted sample may be attributed to reduction of antinutritional factors
such as enzymes inhibitors, this observation was agreed with that reported
by Bradbury and Holloway (1988).
4.7 Effect of processing on protein fractions of defatted pigeon pea

seeds:
The protein fractions (albumins, globulins, prolamins, glutelins and
insoluble proteins) according to solubility were shown in Table 3.
Albumin fraction (which was the major fraction) of untreated sample was
found to be 42.130%, dehulling of the seeds had significantly (p ≤ 0.05)
affected albumin content and increased it to 44.795%, cooking
significantly (p ≤ 0.05) reduced albumin content to 38.765%. However,
cooking followed by dehulling significantly (p ≤ 0.05) reduced it to
40.875%. Reduction of albumin after cooking may be due to
denaturation followed by coagulation of protein by heat treatment, this
result agree with that obtained by Nugdallah (2003) who reported that the
effect of cooking was observed for protein fractions of cowpea. Globulin
fraction (which was the second major fraction) of untreated samples was
found to be 22.623%, dehulling of the seeds significantly (p ≤ 0.05)
affected globulin content and increased it to 25.953%, cooking and
37
100
90
80
In vitro protein digestibility (%)
70
60
50
40
30
20
10
0
Untreated seeds Dehulled seeds Cooked seeds Cooked dehulled
seeds
.
Figure 1. In vitro protein digestibility of processed pigeon pea seeds.
38
70
60
In vitro protein digestibility (%)
50
40
30
20
10
0
Untreated seeds Roasted seeds
Figure 2. In vitro protein digestibility of untreated and roasted pumpkin

seeds.
39
cooking followed by dehulling significantly (p ≤ 0.05) reduced globulin
content to 13.355% and 17.210%, respectiviliy. The results obtained
after processing agree with that obtained by Nugdallah (2003) who
observed an effect of cooking on protein fractions of cowpea. Prolamin
fraction of untreated sample was found to be 3.401%, all treatments used
had no significant (p ≤ 0.05) effect on prolamin fraction which reduced it
to 2.260% after the seeds were cooked and 2.325% after the seeds were
cooked and dehulled. However, dehulling alone increased it to 3.652%.
Glutelin fraction of untreated sample was found to be 15.921%, some
treatments applied had a significant (p ≤ 0.05) effect on glutelin fraction
which was increased to 16.420% after dehulling, and decreased to
10.511% and 12.115% after cooking and cooking followed by dehulling,
respectively. Residue of untreated sample was found to be 13.101%. All
treatments applied significantly (p ≤ 0.05) increased residue to 17.470%,
30.107% and 33.515% after being dehulled, cooked and cooked dehulled,
respectively.
4.8 Effect of roasting on protein fractions of defatted pumpkin seeds:

As shown in Table 3, albumin fraction of untreated sample was found to
be 19.495%, roasting significantly (p ≤ 0.05) reduced albumin to
16.745%. Globulin fraction of untreated sample was found to be
60.521%, roasting significantly (p ≤ 0.05) decreased globulin to 48.490%.
Prolamin fraction was found to be 2.675%, roasting significantly (p ≤
0.05) decreased it to 1.973%. Glutelin fraction of untreated sample was
found to be 3.815%, roasting significantly (p ≤ 0.05) increased it to
5.516%. Residue was found to be 21.440%, roasting had no significant
(p ≤ 0.05) effect on it which was slightly increased to 26.722%. The
result obtained for albumin after roasting disagree with that obtained by
Giami (2004) who reported that albumin content increased during
40
Table 3: Protein fractions (%) of defatted pigeon pea and
pumpkin seeds
Albumin Globulin Prolamin Glutelin Residue

Treatments

42.130(±0.51)b 22.623(±0.48)b 3.401(±0.61)a 15.921(±4.43)ab 13.101(±0.51)d
Untreated
Dehulled 44.795(±1.24)a 25.953(±0.54)a 3.652(±0.74)a 16.420(±0.61)a 17.470(±0.00)c
Cooked 38.765(±0.56)c 13.355(±1.38)d 2.260(±0.86)a 10.511(±0.00)b 30.107(±0.56)b
Cooked 40.875(±0.49)bc 17.210(±1.34)c 2.325(±0.54)a 12.115(±1.10)ab 33.515(±0.70)a

dehulled
(B) Pumpkin seeds:

a a a
Unroasted 19.495(±1.51) 60.521(±11.94) 2.675(±0.25) 3.815(±0.42)b 21.440(±6.29)a
Roasted 16.745(±0.22)b 48.490(±1.70)b 1.973(±0.64)b 5.516(±0.87)a 26.722(±2.96)a

41
fermentation, this difference may be due to the difference between
treatments.
4.9 Effect of processing on digestibility of protein major fractions of

defatted pigeon pea seeds:
The digestibility of major protein fractions of treated and untreated
samples are shown in Table 4. Albumin digestibility of untreated sample
was found to be 69.3755%, all treatments had significantly (p ≤ 0.05)
affect albumin digestibility and it was increased to 82.145% after
dehulling, 99.40% after cooking and 99.96% after cooking followed by
dehulling. Globulin digestibility of untreated sample was found to be
45.085%, dehulling had no significant (p ≤ 0.05) effect on globulin
digestibility and was slightly increased to 47.500%, other treatments
significantly (p ≤ 0.05) affect globulin digestibility and increased to
59.165% after cooking and to 62.590% after cooking followed by
dehulling. The results obtained in this study agree with that obtained by
Coelho and Sgarbieri (1995) who did not observe any reduction in
digestibility of beans after heating, also agree with Genovese and Ljolo
(1997) who reported that the in vitro protein digestibility of bean protein
fraction was low when it was in the native state and was greatly affected
by denaturation, also Anibaba and Osagia (1997) found a continuous rise
in albumin and globulin as result of malting of sorghum.
4.10 Effect of roasting on digestibility of protein major fractions of

defatted pumpkin seeds:
The digestibility of major protein fractions of treated and untreated
sample is shown in Table 4. Digestibility of albumin fraction of
untreated sample was found to be 43.750%, roasting had significantly
(p ≤ 0.05) affect albumin content and increased it to 63.975%. Globulin
42
digestibility of untreated sample was found to be 63.975%, roasting
significantly (p ≤ 0.05) affect globulin digestibility and increased it to
83.960%. The result obtained in this study agree with those of bean seeds
obtained by Colho and Sgarbieri (1995) and Genovese and Lajolo (1997).
43
Table 4: Digestibility (%) of defatted pigeon pea and pumpkin
seeds protein major fractions.
Treatments Albumin Globulin
(A) Pigeon seeds:
Untreated 69.3755(±6.19)c 45.085(±3.42 )b
Dehulled 82.145(±2.52)b 47.500(±0.00)b
Cooked 99.400(±0.13)a 59.165(±8.25)a
Cooked 99.960(±0.04)a 62.590(±11.65)a

dehulled
(B) Pumpkin seeds:
Unroasted 43.750(±8.84)b 63.975(±7.28)b
Roasted 63.975(±7.28)a 83.960(±2.06)a

44
CONCOLUSIONS:
* The processing methods applied in the present investigation resulted
in reduction of antinutritional factors (phytic acid, tannin and
polyphenols).
* Quantitative reduction in antinutritional factors was found to be
accompanied by improvement in protein availability as evidenced by
in vitro protein digestibility.
* Albumin plus globulin fractions of pigeon pea decreased
significantly (P ≤ 0.05) after cooking of the seeds. The treatments
applied had no significant (P ≤ 0.05) effect on prolamin and glutelin
of pigeon pea fractions. On the other hand, the results indicated that
roasting significantly (P ≤ 0.05) decreased albumin, globulin,
prolamin fractions and significantly (P ≤ 0.05) increased glutelin
fraction.
* The results indicated that the digestibility of albumin and globulin
fractions increased significantly (P ≤ 0.05) for all treated pigeon pea
and pumpkin samples.
* Pumpkin seeds compared to pigeon pea was found to be very rich in
protein content. However pigeon pea protein was found to be
available.
RECOMMENDATION:
Due to high digestibility of protein and protein major fractions after
processing of pigeon pea and pumpkin seeds, pigeon pea seed flour
can be used as food in general, as weaning foods to improve the
nutritional quality. Pumpkin seed flour can be used as protein
supplement in some foods. More research should be conducted to
cause more reduction in antinutritional factors and improvement in
IVPD of pigeon pea and pumpkin flour.
45
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Nutritional Evaluation of Processed Pigeon Pea and Roasted Pumpkin Seeds

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NUTRITIONAL EVALUATION OF PROCESSED

PIGEON PEA AND ROASTED PUMPKIN SEEDS

Rania Abbas Ahmed Abusham

A dissertation submitted to University of Khartoum in partial

DEPARTMENT OF FOOD SCIENCE AND

To my family especially my parents

to those whom I will never forget.

I would like to express my deepest thanks to my supervisor Prof.

Elfadil Elfadl Babiker for supporting, advising and encouraging me

and fruitful criticism from which I benefit much.

My deep gratitude to all staff members of the Department of

Food Science and Technology, Faculty of Agriculture, University of

Khartoum, for their help.

My thanks to my family, friends and colleagues, who were ready

to render any assistance I ask for to complete this work.

‫ﻟﻜل ﻤﻥ ﺒﺫﺭﺘﻰ ﺍﻟﻠﻭﺒﺎ ﺍﻟﻌﺩﺴﻰ ﻭﺍﻟﻘﺭﻉ‪.‬‬

CHAPTER ONE: INTRODUCTION …………………………1

CHAPTER FOUR: RESULTS AND DISCUSSION …………… .31

Rania Abbas Ahmed Abusham

A dissertation submitted to University of Khartoum in partial

DEPARTMENT OF FOOD SCIENCE AND

Legumes are valuable agricultural crops because of their capacity to

2.1 Legumes as source of proteins:

2.2 The importance of oil seeds as a source of protein:

2.3 Pumpkin seeds as a source of oil and protein:

2.4 Chemical composition of pigeon pea and pumpkin seeds:

2.4.1 Moisture Content:

2.4.2 Crude protein content:

2.4.3 Fat content:

2.4.4 Crude fiber content:

2.4.5 Ash content:

2.4.6 Carbohydrate content:

2.5 The nutritional value of seeds:

2.6 Effect of processing on nutrients:

2.7 Antinutritional factors:

2.7.1 Phytic acid of pigeon pea and pumpkin seeds:

2.7.2 Tannins content of pigeon pea and pumpkin seeds:

2.7.3 Polyphenols of pigeon pea and pumpkin seeds:

2.8 Effect of processing on antinutritional factors:

2.9 In vitro protein digestibility (IVPD) of pigeon pea and pumpkin

2.10 Effect of processing on In vitro protein digestibility (IVPD):

2.11 Protein quality of seeds:

2.12 Protein fractions of pigeon pea and pumpkin seeds:

2.12.1 Albumins and globulins:

2.13 Effect of processing on protein fractions:

2.14 Digestibility of protein major fractions:

3.2.2 Defatting of pigeon pea and pumpkin seeds:

3.2.3.2. Determination of anti-nutritional factors:

3.2.3.2.2. Determination of phytic acid:

3.2.3.2.3 Determination of polyphenols:

3.2.3.3 In vitro protein digestibility determination:

3.2.4 Physiochemical properties:

3.2.4.1.1 Determination of water–soluble proteins:

Total albumin = T x 14x D.F x 6.25 x N

Albumin % = Total albumin x 100

3.2.4.1.2 Determination of salt soluble proteins:

Total Globulin = T x 14x D.F x 6.25 x N

Globulin % = Total globulin x 100

3.2.4.1.3 Determination of alcohol soluble proteins:

3.2.4.1.4 Determination of alkali-soluble proteins:

Total Glutelin = T x 14 x D.F x 6.25 x N

3.2.4.1.5 Protein content of insoluble fraction:

3.2.5 Statistical analysis:

4.1 Effect of processing on the proximate composition of defatted