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Home , Phaseolus vulgaris, Prosopis, Vicia faba

NUTRITIONAL EVALUATION OF COOKED FABA ( faba L.) AND WHITE BEAN ( vulgaris L.)

By

Sarah Ahmed Elmustafa Abusin B.Sc (Agric).Honours – (2003) University of Khartoum

SUPERVISOR: Prof. Elfadil Elfadl Babiker

A dissertation submitted to the University of Khartoum in partial fulfillment of requirements of the degree of Master of Science in Food Science and Technology

DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY FACULTY OF UNVERSITY OF KHORTOUM

May (2007)

DEDICATION

To my family with affection and gratitude

Sarah

i

ACKNOWLEDGEMENT

Thanks for Allah the Gracious, the great for helping me going through and

finishing this work. My profuse thanks are extended to my supervisor Prof.

Elfadil Elfadl Babiker for supporting advising and encouraging me

throughout the course of this study and thanks for his personal giddiness and fruitful criticism from which I benefit much. Thanks are also extended to Dr.

Gammaa A. Osman for his continuous interest and useful suggestions

Lastly, but not least, sincere appreciation and much thanks are offered to my

mother, father, sister and brothers who fully financed my research, and deep

thanks to friends and colleagues of M.sc.

ii ABSTRACT

Two faba bean cultivars (BB7 and Hudieba-93) and two white bean cultivars (Giza3 and RO21) were used in this study The were cooked. For both cooked and uncooked, the proximate composition, anti nutritional factors and digestibility of the total and protein fractions were determined. The results obtained shows that the moisture content varied between the cultivars and was found to be ranged from 3.4% to 4.07% , content from 0.48% to 1.60% ,crude fiber from 6.5% to 11.9%, ash content from 3.37%to 4.90%, protein content from 19.83% to 31.83% and from 52.96%to 67.43% for both faba bean and white bean cultivars. Cooking of the cultivars seeds caused significant (P ≤0.05) change in the proximate composition, some parameter increased while others decreased. content of uncooked faba bean cultivars was found to be 0.07 and 1.12% for BB7 and Hudieba93, respectively. Phytate was 139.09 and 183.6 mg/100g for the cultivars, respectively while polyphenols were 321.77 and 331.16 mg/100g for the cultivars, respectively. Cooking of faba bean significantly reduced the anti nutritional factors (tannin, phytate and polyphenol). The in vitro protein digestibility of faba bean and white bean cultivars was greatly improved after cooking with maximum value of 89.66% obtained for faba bean BB7 and minimum value of 48.10% obtained for white bean cultivar RO21. Cooking of faba and white bean cultivars was found to improve the metabolism of albumin and glutelin.

iii ﺨﻼﺼﺔ ﺍﻷﻁﺭﻭﺤﻪ

ﺘﻤﺕ ﺩﺭﺍﺴﺔ ﺃﺜﺭ ﺍﻟﻁﺒﺦ ﺍﻟﻤﻨﺯﻟﻲ ﻋﻠﻰ ﻋﻴﻨﺘﻴﻥ ﻤﻥ ﺍﻟﻔﻭل ﺍﻟﻤﺼﺭﻱ ﻫﻤﺎ BB7 ﻭ ﺤﺩﻴﺒﻪ 93 ﻭﻋﻴﻨﺘﻴﻥ

ﻤﻥ ﺍﻟﻔﺎﺼﻭﻟﻴﺎﺀ ﺍﻟﺒﻴﻀﺎﺀ ﻫﻤﺎ ﺠﻴﺯﺍ 3 ﻭ RO21. ﻓﻲ ﺍﻟﻌﻴﻨﺎﺕ ﺍﻟﻤﻁﺒﻭﺨﻪ ﻭﻏﻴﺭ ﺍﻟﻤﻁﺒﻭﺨﻪ ﺍﺠـﺭﻱ

ﺍﻟﺘﺤﻠﻴل ﺍﻟﺘﻘﺭﻴﺒ ﻲ، ﺘﺤﺩﻴﺩ ﻨﺴﺒﺔ ﻤﻀﺎﺩﺍﺕ ﺍﻟﺘﻐﺫﻴﻪ ﻭﺘﻘﺩﻴﺭ ﻨﺴﺒﺔ ﺍﻟﻬـﻀﻡ ﺍﻟﻤﻌﻤﻠـﻲ ﻟﻠﺒـﺭﻭﺘﻴﻥ ﺍﻟﻜﻠـﻲ

ﻭﻷﺠﺯﺍﺀ ﺍﻟﺒﺭﻭﺘﻴﻥ . ﺍﻅﻬﺭﺍﻟﺘﺤﻠﻴل ﺍﻟﺘﻘﺭﻴﺒﻲ ﻟﻸﺭﺒﻊ ﻋﻴﻨﺎﺕ ﺍﻥ ﻨﺴﺒﺔ ﺍﻟﺭﻁﻭﺒﻪ ﻤﺨﺘﻠﻔﻪ ﺒﻴﻥ ﺍﻟﻌﻴﻨـﺎﺕ ﻭ

ﺘﺘﺭﺍﻭﺡ ﺒﻴﻥ 4.07-3.4%، ﺍﻟﺩﻫﻭﻥ ﺒﻴﻥ 0.48-1.60%، ﻨﺴﺒﺔ ﺍﻷﻟﻴﺎﻑ ﺍﻟﺨﺎﻡ ﺒﻴﻥ 6.57 %11.96-

ﺍﻟﺭﻤﺎﺩ ﺒﻴﻥ 4.90-3.37%، ﺍﻟﺒـﺭﻭﺘﻴﻥ ﺒـﻴﻥ 31.83-19.83%، ﺍﻟﻨـﺸﻭﻴﺎﺕ %52.96-67.43.

ﺃﻅﻬﺭﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﺍﻟﻁﺒﺦ ﻴﺅﺜﺭ ﺘﺄﺜﻴﺭ ﻤﻌﻨﻭﻱ ﻋﻠﻰ ﺍﻟﺘﺭﻜﻴﺏ ﺍﻟﻜﻴﻤﻴﺎﺌﻲ ﻟﻠﻔﻭل ﺍﻟﻤﺼﺭﻱ ﻭﺍﻟﻔﺎﺼﻭﻟﻴﺎﺀ .

ﻭﺠﺩ ﺍﻥ ﻤﺤﺘﻭﻯ ﺍﻟﺘﺎﻨﻴﻥ ﻓﻲ ﻋﻴﻨﺎﺕ ﺍﻟﻔـﻭل ﺍﻟﻤـﺼﺭﻱ ﻏﻴـﺭ ﺍﻟﻤﻁﺒﻭﺨـﻪ ﺘﺘـﺭﺍﻭﺡ ﺒـﻴﻥ 1120

ﻤﻠﺠﻡ/100ﺠﻡ ﻭ 70ﻤﻠﺠﻡ/100ﺠﻡ ﻟﻠﻌﻴﻨﺎﺕ BB7 ﻭ H93 ﻋﻠﻰ ﺍﻟﺘﻭﺍﻟﻲ . ﺩﻟﹼـﺕ ﺍﻟﻨﺘـﺎﺌﺞ ﻋﻠـﻰ ﺍﻥ

ﺤﻤﺽ ﺍﻟﻔﺎﻴﺘﻴﻙ ﻴﺘﺭﺍﻭﺡ ﺒﻴﻥ 139.09 ﻭ 183.65 ﻤﻠﺠﻡ/100ﺠﻡ ﻟﻠﻌﻴﻨـﺎﺕ BB7 ﻭ H93 ﻋﻠـﻰ

ﺍﻟﺘﻭﺍﻟﻲ. ﻭﻜﺎﻥ ﻤﺤﺘﻭﻯ ﺍﻟﺒﻭﻟﻴﻔﻴﻨﻭل ﺒﻴﻥ 231.77 ﻭ 331.16 ﻤﻠﺠـﻡ /100ﺠـﻡ ﻟﻠﻌﻴﻨـﺎﺕ BB7 ﻭ

H93 ﻋﻠﻰ ﺍﻟﺘﻭﺍﻟﻲ . ﻟﻭﺤﻅ ﺍﻥ ﺍﻟﻁﺒﺦ ﻟﻌﻴﻨﺎﺕ ﺍﻟﻔﻭل ﺍﻟﻤﺼﺭﻱ ﺍﺩﻯ ﺍﻟﻰ ﺍﻨﺨﻔﺎ ﺽ ﻤﻌﻨﻭﻱ ﻓﻲ ﻤﺤﺘﻭﻯ

ﻤﻀﺎﺩﺍﺕ ﺍﻟﺘﻐﺫﻴﻪ (ﺘﺎﻨﻴﻥ,ﺤﻤﺽ ﺍﻟﻔﺎﻴﺘﻴﻙ ﻭﺍﻟﺒﻭﻟﻴﻔﻴﻨﻭل ). ﻭﺠﺩ ﺍﻥ ﺍﻟﻁﺒﺦ ﺍﺩﻯ ﺍﻟﻰ ﺘﺤﺴﻴﻥ ﻨﺴﺒﺔ ﺍﻟﻬﻀﻡ

ﺍﻟﻤﻌﻤﻠﻲ ﻟﻌﻴﻨﺎﺕ ﺍﻟﻔﻭل ﺍﻟﻤﺼﺭﻱ ﻭﺍﻟﻔﺎﺼﻭﻟﻴﺎﺀ ﺍﻟﺒﻴﻀﺎﺀ، ﻜﺎﻨﺕ ﺃﻋﻠﻰ ﻨﺴﺒﺔ ﻟﻠﻬﻀﻡ ﺍﻟﻤﻌﻤﻠـﻲ ﻟﻌﻴﻨـﺔ

ﺍﻟﻔﻭل ﺍﻟﻤﺼﺭﻱ ﺍﻟﺼﻨﻑBB7 89.66% .ﻭﺃﺩﻨﻰ ﻨﺴﺒﺔ ﻫﻀﻡ ﻤﻌﻤﻠـﻲ ﻟﻠﺒـﺭﻭﺘﻴﻥ 48.10% ﻓـﻲ

ﺍﻟﻔﺎﺼﻭﻟﻴﺎ ﺍﻟﺒﻴﻀﺎﺀ ﺍﻟﺼﻨﻑ RO21. ﺍﻟﻁﺒﺦ ﻟﻠﻌﻴﻨﺎﺕ ﺍﺩﻯ ﺍﻟﻰ ﺘﺤﺴﻴﻥ ﻤﻌﺎﻤـل ﻫـﻀﻡ ﺍﻻﻟﺒﻴـﻭﻤﻴﻥ

ﻭﺍﻟﻘﻠﻭﺘﻴﻠﻴﻥ.

iv

LIST OF CONTENT

Content Pages DEDICATION i ACKNOWLEDGEMENTS ii ABSTRACT iii ABSTRACT v CHAPTER ONE 1 INTRODUCTION 1 CHAPTER TWO 4 LITERATURE REVIW 4 2.1 nutritive value of faba bean and white bean 4 2.2 Chemical composition of faba bean and white bean 6 2.2.1 Proximate analysis 6 2.2.1.1 Moisture content 6 2.2.1.2 Ash content: 6 2.2.1.3 Fat content 7 2.2.1.4 Protein content 8 2.2.1.5 Fiber content 8 2.2.1.6 Carbohydrate content 9 2.3. Protien fractionation 9 2.4. Anti-nutritional factors 12 2.4.1 content 13

v 2.4.2 Phytate content 14 2.4.3 Polyphenol content 15 2.5 In vitro protein digestibility (IVPD) 15

Content Pages 2.6. digestibility of the major fractions 16 CHAPTER THREE 17 MATERIALS AND METHODS 17 3.1 Materials 17 3.2 Processing 17 3.2.1 Soaking 17 3.2.2 Cooking 17 3.3 Methods of analysis 17 3.3.1 Proximate analysis 17 3.3.1.1 Moisture content 17 3.3.1.2 Ash content: 18 3.3.1.3 Fat content 18 3.3.1.4 Crude fiber 19 3.3.1.5 Crude protein 19 3.3.1.6 20 3.3.2 Protein fractionation due to solubility 21 3.3.2.1 Determination of -soluble protien 21

3.3.2.2 Determination of soluble 21 3.3.2.3 Determination of alcohol soluble proteins 22 3.3.2.4. Determination of alkali-soluble proteins 22 3.3.2.5 Protein content of insoluble part 23

vi 3.3.3 Determination of tannin content 23 3.3.4 Determination of phytic acid content 24

Content Pages 3.3.5 Total polyphenols 25 3.3.6 In vitro protein digestibility 25 CHAPTER FOUR 27 RESULTS AND DISCUSSION 27 4.1 Chemical composition 27 4.1.1 Moisture content 27 4.1.2 Fat content 27 4.1.3 Fiber content 28 4.1.4 Protein content 28 4.1.5 Ash content 29 4.1.6 Carbohydrate content 30 4.2 Protein fractions 30 4.2.1 Albumin 30 4.2.2 Globulin 32 4.2.3 Prolamin 32 4.2.4 Glotelin 33 4.2.5 Insoluble protein 34 4.3 Ant nutritional factors 34 4.3.1 Tannin content 34 4.3.2 Phytic acid content 36 4.3.3 Polyphenol content 36

vii 4.4 In vitro protein digestibility 37 4.5 Digestibility of the major fractions 39

Content Pages 4.6 Conclusions 41 4.7 Recommendations 41 REFERENCES 42

viii

LIST OF TABLES

Table Pages 1. Proximate composition (%) of cooked and uncooked faba bean () and white bean () cultivars. 31 2 Protein fractions content (%) of cooked and uncooked faba bean (vicia faba) and white bean (Phaseolus vulgaris) cultivars. 35 3.Tannin, phytic acid and polyphenol content (mg/100g) of cooked and uncooked faba bean (vicia faba) and white bean (Phaseolus vulgaris) cultivars. 38 4. Albumin and glutelin digestibility of cooked and uncooked faba bean (vicia faba) and white bean (Phaseolus vulgaris) cultivars. 40

ix CHAPTER ONE INTRODUCTION

1.1 Introduction Protein-calorie malnutrition (PCM) is a major nutritional syndrome affecting more than 170 million pre-school children and nursing mothers in developing Afro-Asian countries. The present trend in population growth indicates that the protein gap may continue to increase in the future unless well-planed measures are taken to tackle the situation. Provision of adequate proteins of animal origin is difficult and expensive. An alternative for improving nutritional status of the people is to supplement the diet with proteins. Attention, therefore, has been directed to the nutritional evaluation of proteins from plant species (Iqbal et al., 2006). (poor man's meat) play an important role in human since they are rich sources of protein, calories, certain minerals and (Deshpande, 1992). In Afro-Asian diets, legumes are the major contributors of protein and calories for economic and cultural reasons. Food legumes are crops of the family leguminosae also called fabacae. They are mainly grown for their edible seeds and thus are also named legumes. Tropical developing countries are facing an increasing demand for protein-rich food due to teeming population, cereal-based diet and scarcity of fertile land (Sadik, 1991; Weaver, 1994). Legumes are an expensive source of proteins with desirable characteristics such as abundance of carbohydrates, ability to lower the serum cholesterol, high fiber, low fat (except oilseeds), high concentration of poly unsaturated fatty acids and long shelf life. In addition to B complex vitamins, minerals and fiber, legumes are also major source of proteins and calories (Rockland and Nishi, 1979). A wide range of processing techniques could improve the protein and

1 digestibility of legumes and therefore their utilization (Alonso et al., 1998; Conan and Carre, 1989; Frias et al., 199; Gujska and Khan, 1991; Vanderpoel, 1990; Wang et al., 1997). However, it is known that certain treatments such as heat processing, could produce, in some conditions, physicochemical changes in proteins, starch and in the other components of seeds affecting their final nutritional properties (Della Valle, Quillien and Gueguen, 1994; Jeunink and Chefiel, 1979). In general, legumes are source of complex carbohydrates, protein and , having significant amounts of vitamins and minerals and high energetic value (Morrow, 1991; Nielsen, 1991; Tharanathan and Mahadevamma, 2003). Protein contents in legume range from 17 to 40% contrasting with 7-13% of cereal and being equal to the protein contents of meats (18-25%), (Genovese and Lajolo, 2001). Field (Vicia faba) contribute substantially to both human and animal nutrition and is considered as protein source. Faba bean is an important cash crop in the Sudan. The crop contributes to human nutrition and it is the for many people in the Sudan, because of its high protein content as well as other essential . The high lysine content has encouraged the use of faba bean as protein supplement for cereal. Fertilizer application significantly increases the protein content of faba bean (Babiker et al., 1995).

Faba bean is the most important legume crop in Sudan; it is consumed for and supper in many parts of the country, especially in the urban areas. Faba bean like other beans is good source of calories, protein, carbohydrate and fiber (El Tinay et al., 1989). Common bean (Phaseolus vulgaris L.) is a traditional food in human diet, low in fat and rich in proteins, vitamins, complex carbohydrates and minerals. Consumption of dry beans has been linked to reduce risk of diabetes, obesity (Geil and

2 Anderson, 1994), heart disease (Anderson et al., 1984) and colon cancer. Legumes consumed throughout the world, are excellent source of proteins (20 – 25%) and carbohydrate (50 – 60%) and fairly good source of minerals and vitamins (Aykroyd and Doughty, 1977). However their wide acceptability is adversely affected by the presence of tannins, saponius and other anti-nutritional substances (Heniges, Weaver and Nelson, 1991; Morrow, 1991; Stanley, 1992).

1.2 Objective

The objective of this study was to investigate the effect of traditional domestic treatment on faba bean and common bean chemical composition, in vitro protein digestibility, protein fractions and digestibility.

3 CHAPTER TWO LITERATURE REVIEW

2.1 Nutritive value of faba bean and white bean: Plant protein play significant roles in human nutrition particularly in developing countries where the average protein intake is less than required. Food legumes form an important part of the human diet, providing a high proportion of proteins, , carbohydrates, dietary fibers, B-group vitamins (Thiamin, riboflavin, niacin), and minerals, worldwide most grown legumes are , peanut, beans, , and (Kadam and Salunkhe, 1989). In many developing countries they are the main source for human and animal nutrition. In the developed countries, legumes have an increasing use in dietetic formulations in the treatment and prevention of diabetes, cardiovascular diseases, cancer of colon, and lowering of blood cholesterol levels (Bran, et al., 1990), which indicate their possible therapeutic value in humans.

Dry grain legumes, however, contain several anti-nutritional factors, such as α-galactosides, trypsin and chymotrypsin inhibitors, phytates and (Vidal-Valverde et al,. 1992).

Some simple and inexpensive processing technique, such as soaking and cooking are highly efficient for the reduction of these anti-nutritional factors and for improving legume organoleptic quality.

Soaking can reduce the level of total , α-galactosides, minerals, phytic acid and proteolytic enzyme inhibitors (Frias et al., 2000; Vidal- Valverde et al., 2002), which can be partly or totally solubilised and eliminated with the discarded soaking solution. Cooking is probably the oldest treatment for making legumes edible. Usually it includes a previous soaking of the seeds and subsequent cooking in water until they become completely soft. Addition of to the

4 soaking and/or cooking media can produce a reduction of the cooking time (Lu et al., 1984; Van Buren, 1986). In general cooking produce denaturalization of proteins and their diffusion to the liquid phase (Haytowitz and Matthews, 1983). Inactivation of heat sensitive factors, such as trypsin inhibitors (Frias et al., 2000), decrease of phytic acid (Lyer et al., 1989; Khalil and Mansour, 1995; Vidal-Valverde et al., 1994) and α-galactoside contents (El Adawy, 2002). Faba beans are widely used in the Mediterranean region as source of protein in both human and animal nutrition (Larralde, 1982). The nutritional value of field bean has been traditionally attributed to its high protein content, which ranges from25 to 35%, despite the imbalance in sulphur amino acids (Santidrian et al., 1981). Most of these proteins are globulins (60%), albumins (20%), glutelins (15%) and prolamins (Cubero and Moreno, 1983). It is also a good source of sugars, minerals and vitamins. Thus, the chemical analysis of this legume reveals a 50 – 60% content of carbohydrate, which mainly constituted by starch, which the proportion of lipids is relatively low at about 1 – 2.5% with oleic and linoleic acid representing about 75% of fats (Mataix and Salide, 1985).

Dry legumes are a rich and inexpensive source of protein and calories for large part of the world's population, mainly in developing countries. The dry bean (Phaseolus vulgaris) has an important place among the legumes of major production and consumption in Africa, , America and (Bourges, 1987; Reyes, Moreno and Paredes-Lopez, 1993; Sathe et al., 1982). Storage protein of common bean, phaseolus sp., consists of 11.5 – 31% albumins and 46 – 81% glublins. Phaseolin, a 7S globulin is the major storage protein of P. vulgaris, accounting for over 50% of the total proteins. It is an oligomeric protein, consisting of three

5 polypeptide subunits, α-, β-, and δ-phaseolin (Remero et al., 1975; Hall et al, 1977; Bollini and Vitale, 1981).

2.2 Chemical composition of faba bean and white bean: 2.2.1 Proximate analysis: Proximate composition provides a good impression of relative nutritive value and allows basis of comparison between different species, plant parts and cultivation conditions.

2.2.1.1 Moisture content:

The determination of moisture content of foodstuff is very important for both commercial and scientific applications. It is so difficult to determine the moisture content accurately (Food Standard Committee, 1979). Ali et al.,(1982) revealed that moisture content of six Vicia faba cultivars grown at Aliab in Northern Sudan ranged from 8.25 to 8.84. However El Tinay et al., (1993) found that moisture content of 15 genotypes of Vicia faba ranged from 5 to 7.1%. El Sheikh et al.,(1999) reported moisture content was 6.34%. El Sayed (1994) found that the moisture content of Faba bean seeds was 4.3% and a value of 6% was reported by El Tinay et al.,(1989). Ahmed (1997) found that faba bean contains 5.44% moisture. Papiti (1970) found that moisture content of faba bean was 9.0%, while 7.6% moisture was reported by Abdul Rahim (2004). Moisture content of white bean was 4.44% as reported by Diaz et al.,(2002). Lopez et al.,(1986) found that moisture content in white bean was 9.0%, and a value of 5.29% was reported by Costa et al.,(2006).

2.2.1.2 Ash content: The ash content of the foodstuff is the inorganic residue that remains after burning the organic matter. Faba bean contains about 3.2-3.6 % ash as reported by El Sheikh et al., (1999). Ahmed (1997) reported that ash in

6 faba bean was 2.81%. Abdul Rahim (2004) found a range from 3.07 to 3.6%. Ali et al., (1982) found a range varying from 2.65 to 3.03% for six Vicia faba cultivars grown at Aliab in Northern Sudan. Analysis of 15 genotypes of vicia faba showed that the total ash ranged from 2.7 to 7.4% (El Tinay et al., 1993). Duke (1981) claimed that ash content of Vicia faba in some cultivars reached 3.6%, and ranged from 3.45 to 4.21% as reported by Siddig (1999). White beans contains range of 6.99%-7.11% ash as reported by Diaz et al., (2002). Costa et al., (2006) found that ash content was 3.8% for raw white bean beans and 4.0% for cooked one, and ranged from 6.0% to 7.4% in raw white bean as reported by Papiti (1970). A⁄ Rahman et al., (2005) found that ash content was 4.5% for white bean.

2.2.1.3 Fat content: Fats (lipids) are heterogeneous compounds, which are classified according to their solubility in organic solvents as chloroform, ethyl ether, petroleum ether or benzene. The range of fat content in faba bean was 1.1% to 2.2% as reported by El Tinay et al., ( 1989), and 0.7% as observed by Ali et al., (1982) Fat content in faba bean was 1.08% as found by El Sheikh et al., (1999). Papiti (1970) found that fat content was 1.40% in faba bean, and 1.61 as reported by Siddig (1999), Welch and Griffiths (1983) reported that fat content was 1.2% .El Sheikh et al., (1999) reported 1.50% fat content in faba bean. Ahmed (1997) found a range from 0.91% to 1.70%, and Vetter (2003) found 3.36% fat for faba bean. White beans contain 2.49% fat for raw white beans and 2.52% for cooked white beans as reported by Costa et al., (2006). Lopez et al., (1986) found that fat content was 2.5% for raw white bean.

7 2.2.1.4 Protein content:

Protein content of foodstuff can easily be estimated by determining the content, and multiplying that by 6.25 (since each 100 grams contain 16 mg nitrogen). Protein content of faba beans ranged from 28.8 to 30.1% was reported by El Tinay et al., (1989). El Sheikh et al., (1999) and El Sayed (1994) obtained 31.8 to 39% and 28.0 to 37.8% respectively for crude protein of faba beans. Papiti (1970) found that Protein content was 33.69. Siddig (1999) reported 28.4% protein content. A⁄ Rahman et al., (2005) reported 29.5% and 30.2% protein content for two faba bean cultivars . Welsch and Griffiths (1983) found a range of 21.9 - 29.1% for two faba bean cultivar. El Sheikh et al., (1999) reported 32.5% protein for faba bean, and 27.5% as reported by Vetter (2003). Protein content ranged from 26.25 to 30.63% as reported by Ahmed (1997). El Fiel et al., (2002) reported 33.4% protein content for faba bean, and 30.89 to 35.14% has been reported by Abdul Rahim ( 2004). Protein content of white bean was 238 g/kg as reported by Alonso et al., (2000). A/ Rahman et al., (2005) found that protein content ranged from 21.3 to 26.8%. White bean protein ranged from 20 to 23.1% as reported by Lopez et al.,(1986). Costa et al.,(2006) found protein content was 20.9% for raw beans and 22.1% for cooked one. Diaz et al., (2002) found range from 17.4 to 19.9% and Rehman and Shah, (2005) found protein content was 25.3% for raw beans and 25.1% for cooked beans.

2.2.1.5 Fiber content:

There are two terms of fiber known, one is dietary fiber which defined as the non starch polysaccharides and lignin that are not digested or absorbed in the human small intestine, other is crude fiber which represent the insoluble organic residue that remains after boiling a

8 defatted sample successfully with dilute sulphuric acid followed by dilute sodium hydroxide and ignited. It consists of cellulose and hemicellulose (Asp, 1987) . Fiber content for faba bean was 5.17 - 8.08% as reported by Abdulrahim (2004) and ranged from from 8.79 to 9.03% as reported by Elsheikh et al., (1999) and vary from 5.7 to 7.4% as reported by A/Rahaman et al., (2005). In white bean fiber content ranged from 2.81 to 3.27% was reported by Onwuliri and Obu (2002) and a value of 8.55% for raw seeds and 6.25% for cooked seeds was shown by Costa et al., (2006).

2.2.1.6 Carbohydrates:

Carbohydrates are group of food stuff which contains carbon, hydrogen and oxygen in their chemical composition. Carbohydrates of faba beans ranged from 42.7 to 48.3% were reported by El Sheikh et al., (1999) and 52.3 to 54.8% were reported by El Tinay et al., (1989).El Sheikh et al., (1999 found that carbohydrate content of 46.8% for faba bean, while 54.2% was reported by A/Rahman et al., (2005), Siddig (1999) found 50.8% carbohydrate in faba bean, and range from 46.80 to 48.12% was reported by Abdul Rahim (2004). Carbohydrate in white bean ranged from 56 to 62% was reported by Papiti (1970). Diaz et al., (2002) found that carbohydrate content ranged from 56 to 64% in white bean. Costa et al., (2006) observed that carbohydrate was 54.3% for raw white beans and 59.9% for cooked white bean. A⁄ Rahman et al., (2005) found carbohydrate ranged from 57% to 62% in cooked white bean.

2.2.2 Protein fractionations: The major portion of protein in beans was in the form of globulins, followed by glotelin and lesser amount of albumins and prolamin (Nikokoyris and Kandylis, 1997). Albumins present in the seed, which

9 are utilized in the germination process. This fraction is richer in methionine and cysteine than the globulin, at last in peas and faba bean (Bailey and Boulter, 1972). El Fiel et al., (2002) found that the major protein fraction of faba bean was the globulin that ranged from 69.5 to 78.1%. Globulins are storage proteins used during germination and form discrete bodies bound to cell membranes (Bailey and Bulter, 1972). The albumin fraction of faba bean ranged from 1.4 to 3.8%. The prolamin fraction of faba bean ranged from 2.1 to 4.1%. The G1-glutelin fraction of faba bean ranged from 0.9 to 2.2% El Fiel et al., (2000) and found that the G2-glutelin fraction of faba bean ranged from 1.9 to 6.1%, G3-glutelin fraction of faba bean ranged from 8.9 to 14.4% and the insoluble protein (residue) of faba bean ranged from 1.8 to 3.4%. The insoluble protein (residue) consists mainly of proteins from previously defined groups, becoming insoluble due to interactions with lipids carbohydrates or polyphenols via oxidation (Landry and Moureaux, 1981). Indouraine et al.,(1993) found that in the sodium phosphate butter (SPB) protein fraction was the highest (83.2% of recovered protein) followed by salt protein fraction 13.75, 2-mercaptoethanol (2-ME) 1.5%, ethanol (0.8%) and sodium dodecyle sulfate (SDS) solution (0.8%) protein fractions. Cowpea protein was fractionated, on the basis of solubility into albumin (71.4%), globulin (11.1%), prolamin (2.20%) and glutelin (11.0%) was reported by Ragab et al., (2004). Wheat bran proteins were fractionated on the basis of solubility into albumin (23.5%), globulin (15.5%), prolamin (8.5%) and glutelin (25.5%) (Idris et al., 2003). Protein from cowpea seeds can be recovered in six solubility fractions. Fraction I contained salt soluble protein globulin was 65.7% and 79.7%. Fraction II contained water soluble protein albumins was 4.0% and 12.3%. Fraction III contained alcohol soluble protein prolamin was 1.4% and 4.0%. Fraction IV contained G1-glutelins was 0.9% and

10 3.0%. Fraction V contained G2-glutelins was 1.4% and 2.9%. Fraction VI contains G3-glutelin was 0.9% and 14.0% and insoluble protein was 0.5% and 3.0% in two cowpea cultivars (Nugdallah and El Tinay, 1997). Osborne (1924) reported that globulins are the major storage proteins of legumes and they require appreciable salt concentration for solubilization and they account for 50 – 70% of the total seed proteins. Alkali soluble protein fraction was 11.0% in legumes (Deshpande and Nielson, 1987). The glutelin fraction in cowpea seeds was 6.4% of the total seed proteins Dhankher et al.,(1990). In six chickpeas cultivars analyzed for their protein fractions glutelins content ranged from 19.38 to 24.4% (Dhankher et al., 1990). Nugdalla (2003) reported that protein fraction of raw seed of cowpea ranged from 87.5 to 89.8% globulin, 3.6 to 4.0% albumin, 4.3 to

4.5% prolamin, 2.3% G1-glutelin, 2.4 – 2.5% G2-glutelin, 4.5 to 4.8% G3- glutelin and 1.2% residue. About 78% of pigeon 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 while Singh et al.,(1981) found 70.4% albumin and globulin together in decorticated seed of pigeon pea. Singh and Jambunathan (1982) found 3.0% prolamin in whole seed of pigeon pea, also they reported 17.4% glutelin in whole seed of pigeon pea, while Singh et al. (1981) obtained 3.1, 19.6% prolamin and glutelin fractions in decorticated seed of pigeon pea respectively. Nimir (1996) found that albumin which was the predominant fraction for all the pigeon pea lines, to be ranged from 63.54 to 66.41%, globulin fraction ranged from 5.48 to 6.38%, prolamin fraction ranged from 1.95 to 2.68%, glutelin fraction ranged from 10.75 to 14.25% and insoluble residue ranged from 9.5 to 9.58%. Faba bean protein recovery was found to be 79.9% by using tap water as a solvent

11 (McCurdy and Knipfel, 1990), while distilled water extracted 70.7 of the total nitrogen from dried bean seeds (Evans and Kerr, 1963). Abdallah (1997) reported that cowpea albumin was about 80.7%, globulin 6.8%, glutelin 42.12% and prolamin 5.2%. El Khalifa and El Tinay (1994) fractionated the protein of two Sudanese sorghum cultivars, using the Mendle-Osborne procedure. They reported that albumins were 11.5% and 10.0%, globulin 8.2% and 4.7%, prolamin 60.2% and 67.9% and glutelin 10.2% and 9.4% for the low and the high tannin cultivars respectively. Nugdallah (2003) found the cooked cowpea seeds fractions were as follows: globulin 15.7%, albumin 2.2%, prolamin 4.5%, G1-glutelin

2.7%, G2-glutelin 5.1%, G3-glutelin 69.3% and residue 5.5%.

2.2.3 Anti-nutritional factors: Many compounds from food legumes have been shown to cause physiological and biochemical effects, such as pancreas enlargement and growth inhibition in various species of animals (Grant, 1989). The presence of anti-nutritional factors such as enzyme inhibitors, hemagglutinin, factors, polyphenols, tannin and phytic acid, inhibit the proteoytic activity of the digestive enzymes such as pepsin and trypsin as well as the availability of minerals (Deshpande and Cheryan, 1984). Polyphenols can form complexes with metal cations through carboxylic and hydroxylic groups and thus interfere with the intestinal absorption of minerals (Valencia et al., 1999). Phytate content in legumes has been involved in reducing bioavailability of minerals and inhibiting the activity of several enzymes (Deshpande and Cheryan, 1984). Phytate is widely distributed in , especially in seeds, with high concentration in mature legumes, cereal grains and oilseeds (Reddy et al., 1982, Donald, 1983). Moreover, the digestibility of legume protein and legume starch is limited by the presence of anti-nutrients (Lajolo et al.,

12 1991) and the utilization of pulses in both human and animals nutrition is restricted by the presence of the aforementioned factors.

2.2.3.1 Tannins: Tannins are complex phenolic compounds with molecular weight in the range 3000 – 20000 Da, classified either as hydrolysable or condensed, based on their structural types and their reactivity towards hydrolytic agents, particularly acids (Haslam,1987). They are a group of compounds composed of 5 – 7 aromatic rings with 12 – 16 phenolic groups in amolecule. Condensed tannins are dimmers, oligomers and polyemrs of flavan-3-ols which upon acidic hydrolysis, produce anthocyanidins. Tannins are one of the compounds that influence the sensoric quality of food. Tannin content of faba beans ranges from 0.20% to 0.46% for uncooked seeds and ranges from 0.10% to 0.25% for cooked seeds as reported by El sheikh et al.,(1999). Alonso et al., (2000) reported 1.95 g e.q. cat Kt-1DM for raw seeds. Elsheikh and Elzidany (1997) reported 0.024% tannin for raw faba bean. Abdulrahim (2004) reported a range of 0.04 – 0.08% for raw and value of 0.02% for cooked faba bean. El Sheikh et al.,(2000) reported that cooking of faba bean seeds reduced tannin content. Tannin content of beans ranges from 5.37 to 28.79 mg/g dry matter for raw seeds and ranges from 3.55 to 21.01 mg/g of dry matter for cooked beans as reported by Shimelis and Rakshit (2007). Brampama and Simard (2003) reported 14.99 mg equivalent/g, and range from 0.22 to 0.30 g/100g was found by Onwuliri and Obu (2002). Alonso et al., (2000) reported 3.59 g e.q. cat Kt-1DM) in white bean. Nugdallah (2003) reported a range of 0.48 – 0.50 g/100g on dry weight for raw seeds and 0.2 – 0.3 g/100g on dry weight for cooked cowpea.

13 2.2.3.2 Phytate: Phytic acid and/or phytate is the principal storage form of phosphate, ubiquitously distributed in plants, particularly in cereal grains and legumes. The effects of phytic acid in human and animal nutrition are related to the interaction of phytic acid with proteins, vitamins and with several minerals, which thereby restricts their bioavailability. Many attempts to reduce phytate have been tried. According to Fretzdorff and Wiper (1986), there was no reduction of phytate content when whole rye or its were cooked at 100oC but at 170oC phytic acid was reduced by 23%. Faba bean contains about 247.07 to 259.21 mg/100g as reported by Abdulrahim (2004) for raw beans and range from 172.90 to 208.77 for cooked beans. A/Rahaman et al., (2005) reported 291.15 and 298.91 mg/100g phytic acid for two faba bean cultivars. Alonso et al., (2000) found phytic acid content of about 21.7 g Kg-1DM. A range from 0.12% to 0.18% was reported by Elseikh (1999). Elsheikh et al., (2000) found 0.12% for raw faba bean seeds and 0.10% for cooked seeds. Phaseolus vulgaris contains about 1.10% phytic acid of raw seed and 0.86% of cooked beans was found by Boccia et al.,(1998). Shimelis and Rakshit (2007) found a range from 17.34 to 24.06 mg/g DM of raw white bean seed and 6.69 to 8.67 mg/g DM for cooked beans. 16.50 mg/g was found by Barampama and Simard (2003). A range from 1.40 to 1.63 g/100g was reported by Onwuliri and Obu (2000). Lopez et al., (1986) observed range from 1.8 to 2.3 g/100g in raw seed and 1.5 to 2.0 g/100g in cooked white bean. Rehman and Shah (2005) found 1230 mg/100g for raw seeds and 930 mg/100g for cooked seed in white kidney beans. Alonso et al., (2000) found phytic acid content of about 15.9 g/kg DM. A/Rahaman et al., (2005) found 352.51 – 457.69 mg⁄100g for white bean. Nugdallah (2003) found a range from 310.3 to 376.3 mg/100g dry weight in raw

14 seed of cowpea and range from 290.3 to 353.7 mg/100g in cooked cowpea seed.

2.2.3.3 Polyphenols: Polyphenols can form complexes with metal cations through carboxylic and hydroxylic groups and thus interfere with the intestinal absorption of mineral (Valenica et al., 1999). Faba bean contains about 322.08 – 338.64 mg/100g as 0bserved by A/Rahaman et al., (2005). Alonso et al. (2000) found about 3.92 g/kgDM. White bean contains about 2.07 g/kg DM as observed by Alonso et al. (2000). A/Rahaman et al., (2005) found a range from 218.94 to 676.21 mg/100g.

2.3 In vitro protein digestibility (IVPD): The protein digestibility is determined to provide the most satisfactory indication of seed utilization (FAO/WHO/UNV, 1985). Faba bean IVPD have been reported by many researchers, Alonso et al., (2000) reported 70.8% IVPD, El Sheikh et al.,(2000) reported a range of 66.2 – 80.1%. Babiker et al., (1995) reported IVPD of two faba bean cultivars to be ranged from 80.7 to 81.5%, Siddig (1999) found IVPD ranged from 89.0 to 93.8%, El Sheikh et al., (1999) found 79.0%, Alonso et al.,(1999) reported 70.8%. El Sheikh et al.,(2000) obtained IVPD ranged from 66.2% to 80.1% for raw faba bean and 74.6% to 84.6% for cooked faba beans. IVPD was 68.62 - 75.09% as found by Abdul Rahim (2004). In vitro protein digestibility of white bean ranged from 65.8% to 67.4% for raw white bean and 75.2% to 76.0 % for cooked one was reported by Lopez et al., (1986). Alonso et al., (2000) obtained IVPD of 68.1%. Shimelis and Rakshit (2007) obtained 65.63%, 71.14% and 80.66% IVPD for three raw white bean cultivars and 73.52%, 78.98% and 90.31% in three cooked white bean cultivars.

15 2.4. Digestibility of the major fractions: The protein fractions digestibility was found to depend on the processing method such as heat. Genovese and Lajolo (2001) observed that when white bean fractions were heated for 15 minute at 121°C, the digestibility by pepsin was changed from 19.1% to 9.2% for albumin and from 14.9% to 13.8% for glutelin . However when the fractions were digested by both pepsin and pancreatin, the digestibility of heated fractions of white bean were changed from 28.8% to 15.3% for albumin and from 42.1% to 39.6% for glutelin.

16 CHAPTER THREE MATERIALS AND METHODS

3.1 Materials: Faba bean (Vicia faba) cultivars (BB7 and Hudieba-93) and white bean (Phaseolus vulagris) cultivars (Giza3 and RO21) were obtained from Hudieba Agricultural Research Station, Sudan. The seeds were carefully cleaned and freed from broken and extraneous matter. The seeds were milled into fine flour to pass a 0.4 mm mesh size screen. All chemicals used in this study are of analytical grade

3.2 Processing: 3.2.1 Soaking: Seeds were soaked in tap water for 8 hours.

3.2.2 Cooking: Seeds were cooked by boiling process (Domestic processing) then cooled and dried at 55°C to mill into fine flour to pass a 0.4 mm mesh size screen.

3.3 Methods of analysis: 3.3.1 Proximate analysis: 3.3.1.1 Moisture content: Moisture content was determined according to the AOAO (1984) as follows: 2 g of sample were weighed using a sensitive balance in clean dry and pre-weighed crucible and then placed in an oven at 105oC and left over night. The crucible was transferred to desiccators and allowed to cool then weighed. Further placements in the oven were carried out until approximately a constant weight was obtained. Moisture content was calculated using the following formula:

17

(W – W ) – (W – W ) MC (%)= 2 1 3 1 X 100 (W2 – W1) Where: MC = Moisture content W1 = Weight of empty crucible W2 = Weight of crucible with the sample W3 = Weight after drying.

3.3.1.2 Ash content: Ash content of the sample was determined according to the method of AOAC (1990) as follows: 2 g of sample were placed in a clean dry pre-weighed crucible, then the crucible with its contents ignited in muffle furnace at about 550oC for 3 h. or more until light gray ash was obtained. The crucible was removed from the furnace to a desiccators to cool and then weighed. The crucible was re-ignited in the furnace and allowed to cool until constant weight was obtained. Ash content was calculated using the following equation:

(W – W ) AC (%)= 2 1 X 100 (Ws) Where: AC = Ash content W1 = Weight of empty crucible W2 = Weight of crucible with ash Ws = Weight of sample

3.3.1.3 Fat content: Fat was determined according to the method of AOAO (1984) using soxhlet apparatus as follows: An empty clean and dry exhaustion flask was weighed, about 2 g of sample was weighed and placed in a lean extraction thimble and covered with cotton wool. The thimble was placed in an extractor. Extraction was carried out for 6 – 8 h with petroleum ether. The heat was regulated to obtain at least 1 siphoning per hour. The

18 residual ether was dried by evaporation. The extraction flask was placed in an oven till it dried completely and then cooled in a desiccators and weighed. The fat content was calculated using the following equation:

(W – W ) FC (%)= 2 1 X 100 (Ws) Where: FC = Fat content

W1 = Weight of extraction flask W2 = Weight of extraction flask with fat Ws = Weight of sample

3.3.1.4 Crude fiber: Crude fiber was determined according to AOAC (1990), 2g of defatted sample were treated successively with boiling solution of H2SO4 and KOH (0.2 and 0.28 N, respectively). The residue was then separated by filtration, washed and weighed and then washed and transferred into a crucible then placed into an oven adjusted to 105oC for 18 – 24 h. The crucible with the sample was weighed and ashed in a muffle furnace at 500oC and weighed. The crude fiber was calculated using the following equation:

(W – W ) CF (%)= 2 1 X 100 (Ws) Where: CF = Crude fiber W1 = Weight of crucible before ashing W2 = Weight of crucible after ashing Ws = Weight of sample

3.3.1.5 Crude protein: The crude protein was determined by using the micro-kjeldahal method according to AOAC (1984) as follows:

19

a) Digestion: 0.2 g of the sample was weighed and placed in small digestion flask (50 ml), about 0.4 catalyst mixture (96% anhydrous sodium sulphate and 3.5% copper sulphate) was added. 3.5 ml of approximately 98% v/w of

H2sO4 was added. The content of the flask were then heated on an electrical heater for 2 hr or till the colour change to blue-green. The tubes were then removed from the digester and allowed to cool. b) Distillation: The digested sample was transferred to the distillation unit and 20 ml of 40% sodium hydroxide were added. The ammonia was received in 100 ml conical flask containing 10 ml of 2% boric acid plus 3 – 4 drops of methyl-red indicator. The distillation was continued until the volume reached 50 ml. c) Titration: The content of the flask were titrated against 0.02 N HC. The titration reading was recorded. The crude protein was calculated using the following equation:

(T-B) x N x 14 x 100 x 6.25 CP (%)= Ws x 1000 Where: CP = Crude protein T = Titration reading B = Blank titration reading N = HCl normality Ws = Sample weight 1000 = To convert to mg

20

3.3.1.6 Carbohydrates Carbohydrates were determined by difference according to the following equation: Carbohydrate = 100 – (Moisture content + ash content + fat content + fiber content + protein content)

3.3.2 Protein fractionation due to solubility: The Mendel-Osborne (1914) technique for protein fractionation was used in this study.

3.3.2.1 Determination of water-soluble proteins: A sample of 3.5 grams was taken from defatted seed flour for fractionation of total protein. To this amount of the flour, 2 volumes of 50 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 10 ml of the solvent were taken for protein estimation according to the micro-kjeldhal method. The following formula was used for calculating percentage of albumin:

T.F x N x TV x 14 x 6.25 x 100 Soluble protein (%) = a x b x 1000 Where: T.F. = Titer Figures N = Normality of HCl TV = Total volume of the a liquor extracted 14 = Each ml of HCl is equivalent to 14 mg nitrogen a = Number of ml of a liquor taken for digestion (10) b = Number of g sample extracted: (2 g)

1000 = To convert from g to mg

21

3.3.2.2 Determination of salt soluble proteins: The insoluble part obtained after extraction of albumin was re- extracted with 2 volumes of 50 ml NaCl (1 M) for 30 minutes with continuous shaking. The mixture was then centrifuged at 3000 rpm for 20 minutes to separate the insoluble part. The extracted liquor was collected in containers. About 10 ml of the liquor were taken for estimation of soluble protein by the micro-kjeldahl method.

T.F x N x TV x 14 x 6.25 x 100 Soluble protein (%)= a x b x 1000

Soluble protein x 100 Soluble fraction (%)= Total protein of sample

3.3.2.3 Determination of alcohol soluble proteins: The insoluble part obtained after extraction of globulin was re- extracted with 2 volume of 50 ml 70% ethanol added to the insoluble part with continuous shaking for 30 minutes in a mechanical shaker. The extracted 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 as follows:

T.F x N x TV x 14 x 6.25 x 100 Soluble protein (%)= a x b x 1000

Soluble protein x 100 Soluble fraction (%)= Total protein of sample

3.3.2.4. Determination of alkali-soluble proteins: The insoluble part obtained after the extraction of prolamin was re- extracted with 2 volumes of 50 ml NaOH (0.2%) for 30 minutes with

22 continuous shaking. The insoluble part was separated by centrifugation at 3000 rpm for 20 minutes.

The extracted liquor was collected and 10 ml taken for nitrogen determination. T.F x N x TV x 14 x 6.25 x 100 Soluble protein (%) = a x b x 1000

Soluble protein x 100 Soluble fraction (%)= Total protein of sample

3.3.2.5 Protein content of insoluble part:

The remaining insoluble part of the sample was dried and 0.2 gram was digested in 3.5 ml sulfuric acid and used for estimation of insoluble nitrogen.

3.3.3 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 0.2 g 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 2 min. and certified at 2500 rpm for 2 min. One milliliter of the supernatant was pipette into a test tube and 5 ml of vannilin-HCl reagent were added. The optical density was read using a colorimeter (Lab system analyzer-9fiters, J. Mitra and Bros. PVt) at 500 nm after 20 minutes incubation at 30oC. A standard curve was prepared expressing the results as catechin equivalents, i.e. amount of catechin (mg/ml) which gives a color intensity equivalent to that given by tannins after correcting for blank.

Calculation:

23 Tannin concentration was expressed as catechin equivalent (C.E) as follows:

C x 10 x 100 C.E. % = 200

Where:

C= Concentration corresponding to the optical density 10 = Volume of extract in ml 200= Sample weight in mg

3.3.4 Determination of phytic acid content:

Phytate of raw and cooked samples was determined according to the method described by Wheeler and Ferrel (1971). One gram of finely ground sample was weighed into a 125 ml conical flask, extracted with 50 ml 3% to TCA (w/v) for 3 hours with mechanical shaking. Then the suspension was centrifuged at 3000 rpm. Ten milliliters aliquots of the supernatant were transferred into 50 ml boiling tubes. Then 4 ml of FeCl=3= (2 mg ferric ions per ml 3% TCA), centrifuged at 3000 rpm for 15 minutes and the clear supernatant was decanted carefully. The precipitate was then washed twice by dispersing well into 25 ml minutes and centrifuged. The precipitate was cautiously dispersed in a few ml distilled water enriched with 3 ml 1.5 N NaOH with mixing. The volume was made approximately to 30 ml with distilled water and heated in a boiling water bath for 30 minutes. The contents of the tube were filtered hot (quantitavely) through filter paper (Whatman No. 1) and the filtrate was discarded. The precipitate was dissolved in 40 ml of 3.2 N HNO (hot) into a 100 ml volumetric flask. The precipitate was washed with distilled water; the washings were collected in the same flask. The contents of the flask were cooled to room temperature (28 – 32oC) and

24 diluted to volume with distilled water. Five milliliters aliquots were transferred to another 100 ml volumetric flask and diluted to approximately 70 ml with distilled water. Then, 20 ml of 1.5 KSCN ( thiocynate) were added, to complete the volume up to mark. The intensity of the colour was immediately assessed (within one minute) using colorimeter (lab system analyzer – 9 filters, J. Mitra and Bros. PVt. Ltd), at 480 nm. A blank probe was run with each set of sample. A standard curve of different Fe (NO3)3 concentrations was plotted to calculate the ferric ion concentration. The phytate from the ferric ion concentration assuming 4:6 : phosphorous molaration.

3.3.5 Total polyphenol content

Polyphenolics present in faba bean seed and white bean samples were estimated using the Prussian blue assay, as described by Price and Butler (1977). Ground sample (60 mg) was extracted with3 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 FeCl3 in 0.1 N HCl for 3 minutes, followed by the timed addition of 3 ml 0.008 M

K3Fe (CN)6. The absorption was read after 10 minutes at 720 nm on a spectrophotometer (Corning, 259). In all cases, tannic acid was used as a reference standard.

3.3.6 In vitro protein digestibility: In vitro protein digestibility of samples were measured accord-ing to the method developed by Saunders et al. (1973), in which is based on double digestive pepsin-pancreatin system was used in the determinations.

25 In 100 ml conical flask, 250 mg materials were suspended in 15 ml of 0.1 N HCl containing 1.5 mg pepsin (1:10000) and incubated at 37oC for 3 hours. The solution was then neutralized with 0.5 N NaOH and treated with 4 mg pancreatin 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 37oC for 24 hours. Ten milliliter 10% TCA was added to centrifuged at 5000 rpm for 5 minutes. Five milliliter aliquot from the supernatant were taken and analyzed for nitrogen using the kjeldahl procedure (AOAC, 1984).

Calculations: (T – B) x N x 14 x 100 x TV Protein digestibility (%)= (χ) x a

Where: CP% = Crude protein (%) N = Normality of HCl T = ml of titer B = ml of blank a = Number of ml of aliquot TV = Total volume of the mixture 14 = Equivalent weight of nitrogen 250 = Sample weight in mg

250 x CP% χ = 100 x 6.25

26 CHAPTER FOUR

RESULTS AND DISCUSSION

4.1 Chemical composition

The chemical composition of cooked and uncooked faba bean and white bean cultivars seeds is shown in Table 1.

4.1.1 Moisture content:- As shown in Table 1, the moisture content of faba bean cultivars was 3.92% for BB7 and 4.07% for H93.The values obtained in this study were within the range obtained by Elsayed (1994). These values disagree with those obtained by Ali et al., (1982) who reported a range of 8.25 - 8.84% and ElTinay et al., (1993) who reported a range of 5 - 7.1%. After cooking and drying of faba bean the moisture content was found to be 4.77% for BB7 and 4.35% for H.93. These values were agree with those obtained by Nugdallah (2003) who reported a range of 4.8% - 5.2%. The moisture content of white bean cultivars was 3.66% for Giza3 and 3.41% for RO21, and after cooking the moisture content was found to be 3.92% for Giza3 and 3.98% for RO21 cultivar. The values obtained disagree with those reported by Dias et al,. (2002) who found a range of 4.44 - 7.12% in raw seeds.

4.1.2 Fat content:- The fat content of faba bean was 0.81% for BB7 and 1.24% for H93. The results obtained in this study were in agreement with those reported by Eltinay et al., (1989) who reported value of 1.1 - 2.2% and with those reported by Ali et al., (1982) who obtained a range of 0.7% - 1.36% and also with those obtained by Elsheikh et al., (1999) who found a range of 1.08 - 2.12%. After cooking faba bean fat content was found to be 0.48

27 for BB7 and 1.14% for H93.The result obtained were lower than those obtained by Nugdallah (2003) who reported a range of 1.6 - 1.7% for cooked cowpea seeds. For white bean cultivars fat content was found to be 2.28% for Giza3 and 2.13% for RO21. The results obtained agree with those found by Costa et al., (2006) who reported 2.49% and higher than those reported by Papiti (1970) who obtained a range of 1.30 – 1.80%. After cooking fat content of white bean was 1.14% for Giza3 and 1.60% for RO21 which are lower than those obtained by Costa et al., (2006) who reported a fat content of 2.52% in cooked white bean.

4.1.3 Fiber content:- The fiber content of faba bean was 11.96% for cultivar BB7, and 10.88 % for cultivar H93, and after cooking it was found to be 13.86% for BB7 and 13.63% for cultivar H93. The results obtained disagree with those found by Abdurahim (2004) who reported 8.08%, also disagree with those found by Elsheikh et al,. (1999) who found 9.03% fiber, and agree with A/Rahaman et al., (2005) who reported value of 7.4% for uncooked faba bean. For white bean cultivars fiber content was 6.57% for cultivar Giza3, and 7.44% for cultivar RO21, and after cooking it was found to be 7.13% for cultivar Giza3 and 8.53% for cultivar RO21 . Onwuliri and Obu (2002) found that cooking reduce fiber content of white bean and that disagree with the results of this study.

4.1.4 Protein content: The protein content was 29.57% for cultivar BB7 and 31.83% for cultivar H93. The results agree with those obtained by Elsayed et al.,(1994) who reported 28.0 – 37.8% and also agree with those found by Elsheikh et al., (1999) who found 31.8%. The results were higher than those found by Welsch and Griffths, (1983) who found 21.9 - 29.1%, and attributed this wide range to the effect of genetic and environmental factors as reported

28 by Bond et al., (1985). After cooking protein content was found to be 27.67% for cultivar BB7 and 28.15% for cultivar H93. The results were higher than those reported by Nugellah (2003) who found value of 23.6 - 25.9% for cooked cowpea seeds. For white bean protein content was 21.49% for cultivar Giza3 and 19.83% for cultivar RO21. The results agree with those found by Diaz et al., (2002) who obtained 19.9% and also agree with Costa et al., (2006) who reported value of 20.9% for raw white bean. Protein content after cooking was found to be 21.32% for cultivar Giza3 and 18.99% for cultivar RO21 these values were lower than those obtained by Costa et al., (2006) who found value of 22.1% for cooked white bean. In general cooking significantly (P ≤ 0.05) reduce the protein content. That losses in protein could be attributed to partial removal of certain amino acids, along with other nitrogenous compounds, on heating as has been reported by Claweson and Taylor (1993); Monica and Theresia (1992).

4.1.5 Ash content: The ash content of faba bean was 3.47% for cultivar BB7 and 3.37% for cultivar H.93. The results agree with those found by Elsheikh et al., (1999) who reported 3.6%. , Abdul Rahim (2004) who reported 3.6% and Eltinay et al.,(1993) who found value of 2.7 - 7.4% and disagree with those obtained by Rehman and Shah(2005) who reported 2.81% for raw faba bean. After cooking ash content for faba bean was 2.91% for cultivar BB7 and 2.98% for cultivar H93. The results were lower than those found by Nugdallah (2003) who reported value of 4% for cooked cowpea seeds. The ash content of white bean was 4.90% for cultivar Giza3 and 4.60% for cultivar RO21.The results agree with those obtained by Rehman and Shah (2005) who found 4.4%, and lower than those found by Diaz et al., (2002) who found 6.99 and higher than those found by Costa et al., (2006) who found ash content of 3.8% for raw white

29 bean. After cooking ash content of white bean was found to be 3.45% for cultivar Giza3 and 4.42% for cultivar RO21. The results were agree with those obtained by Costa et al., (2006) who found value of 4% for cooked white bean.

4.1.6 Carbohydrate content: The carbohydrate content of faba bean cultivars was 54.60% for cultivar BB7 and 52.96% for cultivar H93. The results were agree with those found by Eltinay et al., (1989) who reported value of 52.3 - 54.8%, and A/ Rahman et al., (2005) who found 52.3 - 54.2% and higher than that found by Abdul Rahim (2004) who found 48.12% for raw faba bean. After cooking faba bean, carbohydrate content was found to be 54.76% for cultivarBB7 and 54.07% for cultivar H93. For white bean cultivars carbohydrate content was found to be 61.26% for cultivar Giza3 and 67.43% for cultivar RO21. The results agree with those obtained by Diaz et al., (2002) who obtained value of 56% - 64% and also agree with those obtained by A/ Rahman et al., (2005) who reported 57.8% - 62.7% for raw seeds. After cooking white bean carbohydrate content was found to be 65.82% for cultivar Giza3 and 65.14% for cultivar RO21. The results were higher than those found by Costa et al., (2006) who reported value of 59.9% for cooked white bean.

4.2 Protein fractions: Table 2. shows the protein fraction of cooked and uncooked faba bean and white bean seeds.

4.2.1 Albumin content: In raw faba bean the major protein fraction obtained was albumin which represent 54.13% for cultivar BB7 and 50.35% for cultivar H93. For white bean the albumin was 66.05% for Giza3 and 68% for Ro21.

30

Table 1. Proximate composition (%) of cooked and uncooked faba bean (Vicia faba) and white bean (Phaseolus vulgaris) cultivars.

Cultivars Treatment Chemical composition Moisture Fat Fiber Protein Ash Carbohydrate

Vicia faba b a b b b b Uncooked 3.92(±0.02) 0.81(±0.00) 11.96(±0.03) 29.57(±0.00) 3.47(±0.03) 54.60(±0.00) BB7 a b a a a ) a cooked 4.77(±0.00) 0.48(±0.01) 13.86(±0.02) 27.67(±0.02) 2.91(±0.06) 54.76(±0.02 b H.93 4.07(±0.01) a b b b b Uncooked 1.24(±0.03) 10.88(±0.02) 31.83(±0.03) 3.37(±0.04) 52.96(±0.07)

a 4.35(±0.02) a a a a a cooked 1.14(±0.01) 13.63(±0.00) 28.15(±0.02) 2.98(±0.02) 54.07(±0.01)

b Phaseolus vulgaris 3.66(±0.02) a b b b b Uncooked 2.28(±0.01) 6.57(±0.01) 21.49(±0.00) 4.90(±0.01) 61.26(±0.03) Giza 3 a 3.92(±0.02) b a ) a a a cooked 1.14(±0.01) 7.13(±0.01) 21.32(±0.00 3.45(±0.03) 65.82(±0.10)

b Ro 21 3.41(±0.00) a b b b b Uncooked 2.13(±0.00) 7.44(±0.01) 19.83(±0.00) 4.60(±0.00) 67.43(±0.02)

a 3.98(±0.02) b a a a a Values cooked 1.60(±0.01) 8.53(±0.01) 18.99(±0.01) 4.42(±0.01) 65.14(±0.01) are means (± SD) Values not sharing a common superscript in column significantly (P≤ 0.05) different.

31

The values obtained were lower than those found by Rageb et al., (2003) who found 71.4% albumin in cowpea protein and agree with those reported by Singh et al., (1981) who found 70.4% albumin and globulin together in decorticated seeds of pigeon pea, and found by Nimir (1996) who reported that the predominant fraction in pigeon pea was albumin which represent 63.54%, and agree with Abdallah (1997) who reported the major protein fraction in cowpea was albumin. After cooking the albumin was found to be 22.91% for BB7 and 25.85% for H93. For cooked white bean the albumin was 29.55% for Giza3 and 26.25% for RO21. Cooking was observed to decrease albumin significantly (P ≤ 0.05).

4.2.2 Globulin:

For faba bean cultivars the globulin content was 20.24% for cultivar BB7 and 18.08% for H93 while for white bean it was 16.5% for Giza3 and 10.65% for RO21 cultivar. The results obtained were higher than those found by Abdallah (1997) who reported a value of 6.8% for cowpea and agree with those found by Indouraine et al., (1993) and Ragab et al., (2003) who reported values of 13.75% and 11.1%, respectively. Cooking of the seeds greatly reduced the globulin content to 4.29% for cultivar BB7 and 4.70% for cultivar H93 while for white bean it was 7.07% for Giza3 and 6.64% for cultivar Ro21. The result obtained disagree with those obtained by Nugdallah (2003) who reported value of 16.4% for cooked cowpea.

4.2.3 Prolamin:- The prolamin content of faba bean cultivars was 2.51% for cultivar BB7 and 2.24% for cultivar H93 while for white bean it was 6.00% for cultivar

32 Giza3 and 5.92% for cultivar RO21. The values obtained were similar to those reported by Elfiel et al., (2000) who reported a range of 2.1 - 4.1%, and also agree with Nugdallah (2003) who found 4.5% for cow pea and Abdallah (1997) who reported a value of 5.2% for cowpea seeds. Prolamin for faba bean after cooking was 2.09% for cultivar BB7 and 3.26% for cultivar H93 while for white bean was 3.59% for Giza3 and 3.26% for RO21.The values obtained were lower than those reported by Nugdallah (2003) who found value of (4.3- 4.5%) for cowpea seeds. The result indicated that, for both faba bean and white bean cultivars, cooking significantly (P ≤ 0.05) reduced prolamin content.

4.2.4 Glotelin:- The glotelin content of faba bean cultivars was 19.33% for cultivar BB7 and 18.15% for cultivar H93 while for white bean was found to be 14.38% for cultivar Giza3 and 10.44% for cultivar RO21. The results obtained agree with those reported by Elfiel et al., (2000) who found that

G1-gluteln of faba bean ranged from 0.9 to 2.2%, G2-glutelin from 1.9 to

6.1% and G3-glutelin from 8.9 to 14.4%, and agree with Dhankher et al., (1990) who reported a range of 19.38 - 24.4% in chick pea, and Singh and Jambunathan (1982)who found 17.4%, the result were similar to that found by Singh et al.,(1981) who found 19.6% and agree with Nimir (1996) who found value of 10.75% - 14.25% for pigeon pea. The results were higher than those found by Ragab et al., (2003) who reported value of 11% in cow pea. Cooking was found to increase glutelin in legumes. Glotelin for cooked faba bean cultivars was 51.54% for cultivar BB7 and 52.50% for cultivar H93 while for white bean was 44.30% for cultivar Giza3 and 53.90% for RO21. The values obtained were lower than those obtained by Nugdallah (2003) who reported value of G3-glutelin as 2.7%,

G2-glutelin as 5.1% and G3-glutelin as 69.3% for cooked cowpea.

33

4.2.5 Insoluble protein :- The insoluble protein of faba bean was found to be 3.60% for cultivars BB7 and 4.7% for cultivar H93, while that for white bean was 2.01% for cultivar Giza3 and 2.95% for RO21. The values obtained agree with those reported by Elfiel et al., (2000) who found a range of 1.8% - 3.4% for faba bean and lower than those reported by Nimir (1996) who found a range of 9.5 - 9.58% for pigeon pea. Cooking of faba bean increased insoluble protein to 17.50% for cultivar BB7 and 19.50% for cultivar H93 and that of white bean to 11.20% for Giza3 and 12.85% for RO21.

4.3 Anti nutritional factors:- 4.3.1 Tannin content: Table 3. shows the effect of cooking treatment on tannin content of faba bean and white bean cultivars. For faba bean cultivar BB7 tannin content was found to be 1120 mg /100g for raw beans and after cooking it was decreased to 50mg/100g, while for cultivar H93 it was 70 mg/100g and reduced to 50 mg/100g after cooking. The values obtained agree with those obtained by Abdulrahim (2004) who reported a range of 0.04 - 0.08% for raw seeds of faba bean. For white bean cultivar Giza3 tannin content was found to be 20 mg/100g for raw beans, and after cooking it was decreased to 10, while for cultivar RO21 it was 40 mg/100g and reduce to 10 mg/100g after cooking. The values obtained agree with those obtained by Elsiddig et al., (1997) who reported a range of 0.02 - 0.03% and disagree with values obtained by Onwuliri and Obu who reported 0.22 - 0.30% for raw white bean. Elsheikh et al., (2000) reported that cooking significantly (P < 0.05) decreases tannin content of faba bean Nugdallah (2003) found that cooking significantly decreases tannin content of cowpea.

34 Table 2. Protein fractions content (%) of cooked and uncooked faba bean(Vicia faba ) and white bean (Phaseolus vulgaris) cultivars.

Cultivars Treatment Protein Fractions

Albumin Globulin Prolamin Glotelin Insoluable Recovered Vicia faba b ) b b b b b Uncooked 54.13(±0.00) 20.24(±0.00 2.51(±0.01) 19.33(±0.03) 3.60(±0.00) 99.80(±0.03) BB7 a ) a a a a a Cooked 22.91(±0.21) 4.29(±0.02 2.09(±0.00) 51.54(±0.01) 17.50(±0.00) 98.33(±0.24) b b b b b ) b H.93 Uncooked 50.35(±0.25) 18.08(±0.03) 2.24(±0.01) 18.15(±0.01) 4.70(±0.00) 93.51(±0.21 )a a a a a ) a Cooked 25.85(±0.04 4.70(±0.20) 3.26(±0.05) 52.50(±0.00) 19.50(±0.00) 105.80(±0.19

b ) b b b b ) b P. vulgaris Uncooked 66.05(±0.05) 16.5(±0.07 6.00(±0.05) 14.38(±0.00) 2.01(±0.01) 105.01(±0.08 Giza 3 a a a a a a Cooked 29.55(±0.00) 7.07(±0.00) 3.59(±0.02) 44.30(±0.21) 11.20(±0.10) 95.70(±0.32) b b b b b b RO 21 Uncooked 68.00(±0.00) 10.65(±0.07) 5.923(±0.13) 10.44(±0.08) 2.95(±0.05) 97.96(±0.19) a a a a a a Cooked 26.25(±0.15) 6.64(±0.00) 3.26(±0.00) 53.90(±0.10) 12.85(±0.35) 103.00(±0.50) Values are means (±SD) Values not sharing a common superscript in column significantly (P<0.05) different.

35 4.3.2 Phytic acid content:-

Table 3. shows phytic acid content of faba bean and white bean cultivars, before and after cooking .

For faba bean cultivars BB7 phytic acid was 183.65 mg/100g for raw beans and after cooking it was significantly (P ≤ 0.05) decreased to 153.44 mg/100g. For cultivar H93 it was 139.09 in raw bean while after cooking significantly (P ≤ 0.05) decreased to 104.23mg/100g. The values obtained agree with those reported by Elsheikh et al.,(1999) who found a range of 0.12- 0.18 mg/100g for uncooked seeds of faba bean. Phytic acid content for white bean cultivar Giza3 was 151.83mg/100g in raw bean and after cooking it was significantly (P ≤ 0.05) decreased to 97.1mg/100g, For cultivar RO21 was 234.51mg/100g in raw bean and after cooking significantly (P ≤ 0.05) decreased to143.31mg/100g. The results obtained for raw seeds was lower than those found by A/Rahaman et al., (2005) who found a range of 352.51-457.69 mg/100g for raw seeds of white bean. The results obtained indicated that cooking had a profound effect on phytate content and significantly minimize its amount for both faba bean and white bean cultivars. The results agree with those of Elsheikh et al.,(2000) and, Rehman and Shah (2005) who reported that cooking significantly decreased phytic acid content of faba bean and white bean, respectively.

4.3.3 Polyphenol content:- For faba bean cultivar BB7 polyphenol content was 231.77 mg/100g for raw seeds and after cooking it was significantly decreased to 102.66 mg/100g, and for cultivar H93 was 331.16/100g for raw seeds and after cooking significantly (P ≤ 0.05) decrease to 148.97 mg/100g. The results obtained for raw seeds were agree with those found by A/Rahaman et al.,

36 (2005) who reported 322.08 - 338.64 mg/100g. Polyphenol content for white bean cultivar Giza3 was 646.78 mg/100g for raw seeds and after cooking significantly (P ≤ 0.05) decrease to 428.02 mg/100g. For cultivar RO21 was 234.78 for raw seeds and after cooking significantly (P ≤ 0.05) decrease to 123.53 mg/100g. The results obtained for raw seeds were agree with those found by A/Rahaman et al., (2005) who reported 218.94 to 676.21 mg/100g for white bean. The result obtained indicated that cooking had a profound effect on polyphenol content and significantly minimize its amount for both faba bean and white bean cultivars.

4.4 In vitro protein digestibility (IVPD):- Table 4 shows the IVPD of faba bean and white bean cultivars before and after cooking. For faba bean cultivar BB7 the IVPD was 78.33% for raw bean and after cooking it increased to 89.66%, while for H93 cultivar it was 73.56% for raw bean and after cooking it increased to 76.87%. For white bean cultivar Giza3 the IVPD was 52.77% for raw bean and after cooking it increased to 72.10%, and that of cultivar RO21 was 45.73% in raw bean and it increased to 48.10% after cooking. The results obtained agree with those reported by Elsheikh et al. (2000) who found a range of 66.2% - 80.1% for uncooked faba beans and with those obtained by Abdul Rahim (2004)who reported value of 68.62-75.09% for raw faba bean, and higher than those obtained by Alonso et al.,(2000) who gave a value of 70.8% for raw faba bean. The results obtained for raw white bean agree with those obtained by Rehman and Shah (2005) who gave value of 34.0% and lower than those found by Shimelis and Raksht (2007) who found 65.63 - 80.66% for raw seeds. The results obtained indicated that cooking of legumes significantly affected the in vitro protein digestibility. This could be attributed to inactivation of anti nutritional factors such as tannin and phytate. Elsheikh et al., (2000) and Lopez et al., (1986) reported that cooking increase IVPD in faba bean and white bean.

37

Table 3. Tannin, phytic acid and polyphenol content (mg/100g) of cooked and uncooked faba bean (vicia faba) and white bean( phaseolus vulgaris ) cultivars.

Cultivars Treatment Anti nutritional factors

Tannin Phytic acid Polyphenol

Vicia faba b b b Uncooked 1120 (±0.00) 183.65(±0.00) 231.77(±0.87) BB7 a a a cooked 50(±0.00) 153.44(±0.00) 102.66(±0.18) b b b H.93 Uncooked 70 (±0.00) 139.09(±0.00) 331.16(±0.32) a a a cooked 50 (±0.00) 104.23(±0.00) 148.97(±0.09)

b b b Phaseolus vulgaris Uncooked 20 ±(0.00) 151.83(±0.02) 646.78(±1.08) Giza 3 a a a cooked 10 (±0.00) 97.10(±0.00) 428.02(±0.00) b b b RO.21 Uncooked 40 (±0.00) 234.51(±0.00) 234.78(±0.89) cooked 10 (±0.00)a 143.31(±0.01)a 123.53(±0.60)a Values are means (±SD) Values not sharing a common superscript in column significantly (P<0.05) different.

38 4.5 Digestibility of the major fractions:-

Table 4. shows the in vitro digestibility of Albumin and glutelin of faba bean and white bean cultivars before and after cooking. Cooking of faba bean and white bean significantly (P < 0.05) increased the digestibility of Albumin from 27.81% to 47.5% and from 22.96% to 28.8%,respectively. The results obtained disagree with those obtained by Genovese and Lajolo (2001) who found the heat treatment reduced the digestibility of albumin. Cooking of faba bean and white bean significantly (P < 0.05) increased the digestibility of glutelins from 40.6% to47.5% and from 29.01% to 39.89%, respectevily. The results disagree with those obtained by Genoves and Lajolo (2001) who observed reduction in the digestibility of white bean. Araüjo et al., (2002) found that native globulins are resistant to digestion by isolated enzymes, but hydrolysis by cooking greatly increased.

39

Table 4. Albumin and glutelin digestibility (%) of cooked and un cooked faba bean ( V. faba) and white bean (P. vulgaris)

Cultivars Treatment Total* Albumins Glutelins b b b Vicia faba BB7 Uncooked 78.33 (±0.17) 30.00 (±0.16) 33.71 (±0.06) a a a cooked 89.66 (±0.33) 38.75 (±0.09) 41.67(±0.01) b b b H.93 Uncooked 73.56 (±0.44) 25.62 (±0.01) 47.50 (±0.16) a a a cooked 76.87 (±0.17) 56.25 (±0.03) 53.33 (±0.08)

b b Phaseolus vulgaris Uncooked 52.77 (±0.00) 18.33 (±0.08) 28.03 (±0.08) Giza 3 a a a cooked 72.10 (±0.07) 30.00 (±0.16) 42.25 (±0.01) a b b RO.21 Uncooked 45.73 (±0.42) 17.60 (±0.16) 30.00 (±0.16) a a a cooked 48.10 (±0.40) 27.60 (±0.16) 37.52 (±0.01)

Values are means (±SD) Values not sharing a common superscript in column significantly (P< 0.05) different. * Digestibility of total protein.

40 Conclusions The results obtained in this study indicated that cooking significantly (P < 0.05) increased ash and fiber contents while significantly (P < 0.05) reduced the protein and fat content for both cultivars. Also cooking caused a significant (P < 0.05) decrease in anti-nutritional factors (tannin, phytic acid and polyphenol) with a concomitant increase in IVPD of faba and white bean as well as digestibility of albumin and glutelin.

Recommendation: Due to high protein content and high in vitro protein digestibility for faba bean and white bean and according to the finding it is strongly recommended cooking of beans to a level that could minimize the anti nutritional factors with a concomitant improvement in the nutritional value of the end product.

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