University of Nigeria Research Publications

NWABUGWU, Chika C. A

utho

r

PG/M.Sc/00/28319

Title

Production and Evaluation of Weaning Formula From Pigeon

Pea (Cajanus Cajan) and Millet Seed (Pennisetum Americanum) Blends

Facu

lty

Agricultural Sciences

Dep

artm

ent

Food Science and Technology

Dat

e October, 2005

Sign

atur

e

PRODUCTION AND EVALUATION OF WEANING FORMULA FROM PIGEON PEA (Cajanus cajan) AND

MILLET SEED (Pennisetum americanum) BLENDS.

AN M.SC. PROJECT REPORT: SUBMITTED TO THE DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY,

FACULTY OF AGRICULTURE, UNIVERSITY OF NIGERIA, NSUKKA I N PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF M. SC. I N FOOD

SCIENCE AND TECHNOLOGY

DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY UNIVERSITY OF NIGERIA,

NSUKKA

OCTOBER, 2005

CERTIFICATION

This project report titled "PRODUCTlON AND EVALUATION OF

WEANING FORMULA FROM PIGEON PEA (Cajanus cajan) AND MILLET

SEED (Pennisetum nrnericanum) BLENDS" has becn presented by Nwabugwu,

Chika C. (PG/MSc/00/283 19) of the Department of Food Science and Technology,

Faculty of Agriculture, University of Nigeria, Nsukka.

The work embodied in this project report is original and has not been

submitted in part or in full for any other diploma or degree of this or other

University.

Dr. J. C. Ani (Mrs) (Project Supervisor)

Dr. N. J. Enwere (Mrs) (Head of Department)

I

. . . 111

DEDICATION

This research work is dedicated to

Almighty God

And

My Mother

IV

The successful accomplishn~ent of this work was only possible through the efforts,

goodwill, kindiiess and commitment of several persons whom I musl appreciate.

I sincerely thank my supetvisor; Dr. J. C . Ani (Mrs) for hes dedication, untiring efGorts,

tolerance and guidancc in assuring that this work was successfblly completed. I thank also Prof.

Onwuka, Prof. Ihekuronye, Dr. Uvere and the entire staff of Food Science and Tdmology

Department, U.N. Pa , for their lectures, advice, kindness and support during the period of tlis

study. 1 am lu~hly indebted to Prof: 1. C. Obizoba of Ijotne Sciehce and Nutrition Department,

U.N.N., who through his fatherly advice and supervision made this work LO progress. I extend

my sincere gratitude to Dr. Ani of Animal Science Dcpartmcnt, U.N.N, for his advice and

direction in support of this work.

I acknowledge tlae support given to me by my colleagues; Chinwe Osuagwu, lfeoma

Pvlbacyi, Chinedu Iloanusi, and Obioha Okolie, I also appreciate my friends; Anwuli Baiden,

C11ima Nwambara, Parker Elijah, Charity Umeano and Presbyter Onono&u for their

encouragement to me during this work.

Finally, my sinccre thanks go to my mother and brother Ejike for their financial support

during this study. 'To thc rest of my brothers and sisters for their support and encouragement

during this study Abovc all, I Lhank God who gave me the ability Lo carry out this research

successfi'uly.

Fer~nented (9hrs)- sprouted (24hrs), steamed (30mins), and dried millet and pigeon pea seeds

were nlilled into llours respectively and used in [he Somulation of weaning formula. The

cornposition and fimctional properties 01 the llours were dctennined. Composite blends were

ionnulated with the processed flours to contain 70% crude protein from pigeon pea and 30%

crude prolein froin millet. Casein and a commercial weatling formula served as controls.

Prcfercncc test on the formulated blend was done by ranking and .the preferred products were

subjected to a 35-day animal assay, monitoring growl11 and N balance. Fermentation increased

the crude protein content of pigeon pea by 6.44% and millet by 3.05%. Sprouting increased

protein content of pigeon pea by 13.2% but seduced that of millet by 14.16%. Steaming sprouted

rnillet and pigeon pea reduced the crude protein content to 8.6% and 16.87% while fermented

millct and pigeon pca crude protein were reduced to 7.83% and 18.63% respectively.

Fernlentation reduced the tannin and phytate levels of millet and pigeon pea but increased the

cyanide level. Sprouting caused an increase in the tannin and cyanide level of millet but a

recluction in the tannin, cyanide and phytate level of pigeon pea. Steaming incrc-ascd tannin and

cyanide level of millct but reduced their content in pigcon pea. All treatments increased the water

absorption capacity of millct and pigeon pea flours. The formulated blends showed comparable

crudc protein content will1 the casein diet, 9.53%, and the FA0 standasd protein value of 10%.

The porridgcs prepared from the formulated blends were generally acceptable organoleptically.

Among the most acceptable blends, \511ich were analysed for nutritional quality through animal

assay, diet IV (SSPF + FSMF, 70:30) had comparable food intake (138.248) and digested N

(0.46g) with casein diet (130.798 and 0.48g). From the result, it was inferred that, a foimulated

dict of Ser-mentcd stcamed millet and sprouted steamed pigeon pea, which showed a high

nutritional quality could be an acceptable formula for infants and growing children in a

developing country like Nigeria.

TABLE OF CON'UPJT Title Page -- -- -- --

Certification -- -- -- -- Dedication -- -- -- -- Acknowledgemei\l -- -- -- Abstract -- -- -- -- Table of Contents .-- -- -- Lisl OF Figures -- -- -- List of Tables -- -- -- -- Chapter One ---- - - -- -- 1 0 Introduction ---- -- -- 1.1 Statement of Research-- -- 1 2 Justification for the Study -- --

CHAPTER TWO: LI'FEHATURE -- Ii~troduction -- -- -- Nutritive Value of Cereal -- --

Nutritive VaIue of Legume. --

Anti-nutritional Factors in Legumes --

Antinutritional Factors in Cereals -- -- Detoxification Treatments -- -- --

Use of Cereals and Legipmes as Weaning Foods -- --

Production, Utilization and Nutritive Composition of Millet -- -- -- 18

Production, Utilization and Nutritive Composition of Pigeon Pea. -- -- 20

Compositions and Nutritional Requirement of Weaning Foods -- -- -- 23

Protein Availability in Weaning Food -- -- -- -- -- -- 25

Dietary Bulk in Weaning Foods -- -- -- -- -- -- -- 26

Wcaning Foods and Practices in West Afiica -- -- -- -- -- 27

Previous Studies on Weaning Food Formulation -- -- -- -- -- 28

CHAPTER THREE: MATERIALS AND METHODS -- -- -- -- 30

3.1 Reparation of raw material -- -- -- -- -- -- -- -- 30

vii

3.3 Analytical Methods -- -- -- -- -- -- -- -- 33

3.3.1 Proximate Analysis --- -- -- -- -- -- -- -- -- 33

3.3.1.1 Moisture Content (A.O.A.C., 1990). -- -- -- -- -- -- 3 3

3.3.1.2 Pat Content (AOAC, 1990) ---- -- -- -- -- -- -- 3 3

3.3.1.3 Ash Content (AOAC, 1990) -- -- -- -- -- -- -- 3 5

3.3.1.4 Crude Protein Determination (Nitrogen X 6.25) (AOAC, 1990) -- -- -- 3 5

3.3.1.5 Crude Fibre Determination (AOAC 1990) -- -- -- -- -- ..- 36

3.3.1.6 Nitrogen Free Extract Determination -- -- -- -- -- -- 3 6

3 3 .1 .7 Caloric Value Determination -- -- -- -- -- -- -- 3 7

3.3.1 . 8 Determination of True Protein -- -- -- -- -- -- -- 3 7

3.3.1.9 Mineral Content Determination -- -- -- -- -- -- -- 3 8

3.3.2 Determination of Antinutritional Factors -- -- -- -- -- -- 3 8

3.3.2.1 Determination of Hydrogen Cyanide Level -- -- -- -- -- 3 8

3.3.2.2. Determination of Phytate Level -- -- -- -- -- -- -- 39

3.3.2.3. Deterinillation of Tannin Levels -- -- -.. -- -- -- -- 40

3 3 . 3 . Functional Properties Determination -- -- -- -.. -- -- 40

3 3.3.1 . Particle Size Distribution -- -- -- -- -- -- -- -- 40

3.3.3.2. Viscosity -- -- -- - - -- ---- -- -- -- -- 4 1

3.3.3.3. Water Absorption Index-- -- -- -- ---- -- 3.3.3.4. Water Solubility Index -- -- -- -- -- -- --

3 3.3.5. Least Gelation Concentratiorr- -- -- -- ..-

3.3.3.b. Bulk Density Determination -- -- -- -- -- 3.3.3.7. Reconstitution Time Determination -- -- -- --

3.3.4. Sensory Analysis -- -- -- -- -- -- --

3.3.5 Animal Bioassay -- -- -- -.. -- -- --

3.3.6 Statistical ~na l~s i s - - - -- -- -- -- -- --

4.0 Results and Discussions -- -- -- -- -- -- 4.1 Nutrient Composition -- -- -- -- -- --

4. I . I Effect of Fermentation on Proximate Composition of Millet and

Pigeon Pea Flours -- -- -- -- -- -- --

4.1.2 EfTect of Sprouting on Proximate Composition of Millet and

Pigeon Pea Flours -- -- -- -- -- -- -- -- ECfect of Fermentation on Antinutritional Content ofSPigeon Pea and Millet --

Effcct of sprouting on antinutrient content of millet and pigeon pea -- --

Effect of Steaming on Proximate Composition of Fermented and Sprouted

Millet and Pigeon pea -- -- -- -- -- -- -- --

Effect of steaming on the mineral content of fermented and sprouted

pigeon pea and millet -- -- -- -- -- -- -- -- Effect of stcaning on antinutritional content of fcrmcnted and sprouted

pigeon pea and millet. -- -- -- -- -- -- -- -- Functional Properties -- -- -- -- -- -- -- -.. Effect of Fermentation on Functional Properties of Millet and

Pigeon pea Flour -- -- -- -- -- -- -- -- -- Effect of Sprouting on Functional properties of Millet and Pigeon pea Flour --

Effect of' steaming on functional properties of fermented and sprouted

pigeon pea and millet flour: -- -- -- -- -- -- -- Sensory Evaluation -- -- -- ..- -- -- -- -- Animal Bioassay -- -- -- -- -- -- -- -- -- Proximate Composition of Diets used for Animal Bioassay -- -- -- Mineral Composition of diets used for animal bioassay -- -- -- -- Functional Properties of diets -- , -- -- -- -- -- --

Animal Bioassay -- -- -- -- -- -- -- -- --

viii

49

5 1

5 3

55

5 7

59

60

GO

63

68

70

7 3

7 3

75

7 6

79

CHAPTER FIVE ---- -- -- -- -- -- -- -- -- 83

5.0 Conclusions, Recommendations and Suggestions for Further studies-- -- 83

5.1 Conclusions -- -- -- -- -- -- -- -- -- 8 3

5 2 Recommendations -- -- -- -- -- -- -- -- -- 8 3

5.3 Suggestions for Further Studies -- -- -- -- -- -- -- 84

KIFAXRENCES -- -- -- -- -- -- -- -- -- 8 5

Appendix A: Graphs showing the efl'ect of fermentation and sprouting -- -- 97

Appendix B: Calculation of the diet conlposition fed to rats -- -- -- 103

Appendix C: Scnsory evaluation score sheet -- -- -- -- -- 106

LlST OF FIGURES Use of the food square to plan weaning diets and mixes -- - -- 23

Flow diagram for millet flour production -- -- -.. -- -- 3 1

Flow diagram for Pigeon pea flour production -- -- -- -- -- 32

LISTS OF TABLES

Chemical Composition of major cereal grains (1 00g edible portion) -- -- 6

Energy and selected nutrients in legumes (composition per 100g edible portion of dricd mature whole seeds). -- -- -- -- -- -- --

Examples of Antinutritional factors in plant foods. -- -- -- --

Leading Millc t Producers, 1990-- -- -- -- -- -- --

Leading Millet Producers, 2001 -- -- -- -- -- -- --

Essential amino acid composition ( d g ) and chemical score of millet and sorghum proteins -- -- -- -- -- -- -- --

Leading Pea Producers, 2001 -- -- -- -- -- -- --

Leading Pulses Producers, 200 1 -- -- -- -- -- -- --

Ainino acid con~position ( d l 6gN) of the protein of raw legume seeds -- --

Daily Average Energy and Fluid Requirements and Safe level of Protein Intake lor I n h t s and Children three months to iive years, Sexes combined (FAO/WHO/IJNU, 1983)- -- -- -- -- -- -- -- Proportions of difkrent flour treatrnent in the composite blends -- -- --

Composition on dry weight basis ( d l 00g) of millet, pigeon yea and casein diets fed to rats -- -- _ _ . -- -- -- -- -- --

Effect of fermentation time on proximate composition of millet and pigeon pea flours (% dry weight basis) -- -- -- -- -- --

Effect of sprouting time on proximate composition of millet and pigeon pea flours (% dry weight basis). -- -- -- -.. -- --

Effect of fermentation on antinutritional content of pigeon pea and millet --

Effcct of sprouting on antinutritional content of millet and pigeon pea -- --

Effect of steaming on proximate composition of fermented and sprouted millet and pigeon pea flours (Oh dry weight basis) -- -- -- --

Effect of ste8ming on the mineral content of fermented and sprouted pigeon pea and millet -- -- -- -- -- -- --

Effect of steaming on antinutritional content of fermented and sprouted pigeon pea and millet -- -- -- -- -- -- -- --

Effect of fernlentation on functional properties of millet and pigeon yea flours--

Effect of sprouting on the functiond properties of sprouted millet and pigeon pea flours -- -- -- -- -- -- -- --

Effect of steaming on functional properties of fermented steamed and sprouted steamed millet and pigeon pea flours -- -- -- --

Sensory evaluation scores of the formulated products, commercial diet a d casein dict -- -- -- -- -- -- -- --

Proximate composition of diets used for animal bioassay (% dry weight basis) -- ..- -- -- -- -- -- --

Mineral Composition of diets used lbr animal bioassay -- -- --

Functional Properties of diets used for animal bioassay -- -- -- Food intake, gain in body weight, protein efficiency ratio (PER), N intake, feacal N, urinary I?, digested N, retained N, biological value (BV) and net protein utilization of rats fed diets based on fermented and sprouted millet atld pigeon pea. -- -- -- -- -- -- -- --

CHAPTER ONE

1.0 Introduction

Breast-feeding is considered best for infalts froin nutritional and' immunological points

of view as well as for protection against campylobacter-associated diarrhoea (Nout and Ngoddy,

1997). In developing countries most infants show satisfactory growth for the first four to six

moilths of life when breast milk solely meets the nutritional needs but after this period, it may

become increasingly inadequate as the nutritional demands of the inf&t increases (Nkama et ul.,

2001).

Weaning is a gradual process during which breast milk is increasingly coinpleinented

with other foods that fully meet the growing child's needs. Mitzner et ul., 1984, refers to

weaning process as the transition from a diet of breast milk to a diet that includes breast milk and

other foods and finally to a family diet. It is a process of gradual introduction of foods into a

baby's diet to complement breast milk and progressively replacing it and eventually adapting the

child to adult diets (Nltarna et al., 2001). According to Fashakin (1980), weaning is the

transitional period following lactation and full dependence of the child on non-milk products.

A gradual increase in provision of complementary (weaning) food causes a concomitant

reduction in the child's dependence on breast milk, this reduction continues until the child can

fulfill all his or her nutritional needs with an adult diet. The weaning process could start from

four to six months and varies fiorn one socio-economic status to the other. It could be gradual, t

lasting for months until the infant is fully introduced to the family diet (Onofiok and

Nnanyelugo, 1998). On the other hand, it could be abrupt, in which case the infant is introduced

straight into the family menu. Abrupt weaning creates problems, as the child may not be able to

eat enough of the adult diet to meet his or her nutsitional needs.

In most developing countries, about 50% of all pre-school children are chronically

malno~uiished. According to UNICEF's report on "The state of the world's children, 1989",

covering over 60 countries, more than 150 million children under 5 years of age suffer from

~naln~ltrition, n~ainly of the protein -energy type (PEM). Protein-energy malnutrition stands out

as the most serious of the nutritional deficiency problem in Nigeria as well as in other

economically developing countries of the world and among low income earners in developed

countries (Ossai and Malomo, 1988). The immediate cause of this high rate of malnutrition is the

combination of various diseases and dietary inadequacies interacting in a mutually reinforcing

menner. Young children remain particularly at risk because of their great energy and nutrient

needs and vulnerability to infection.

Second to protein energy malnutrition is anaemia, which is another common nutritional

problem in many countries of the developing world. The most frequent cause of anaemia is low

iron bioavailability from the predominantly cereal- based diet and/ or blood loss due to parasitic

infestation with hookworms and malaria. The low bioavailability of iron from cereal diets is due

to the presence of different inhibitors such as phytate and tannin (Lorri, 1993).

In Nigeria, as in most developing parts of the world, most people depend on plant for

food supplies. Cereals and legumes play important role in the diets of many people including

children, and are the major sources of proteins, calories, vitamins and minerals. Many countries

have employed cheap locally available plant materials from cereals and legumes for the

development of wearling foods (Nkama el al,, 2001). The use of these plant materials by humans

is very limited mainly because of their lower protein quality when compared with animal

products and the anti~lutritional factors which interfere with the utilization of the nutrients

(Onuoha and Obizoba, 200 1).

Cereals are fruits of cultivated grasses also known as caloric or starchy foods, which are

important sources of dietary fibre. Examples include maize, rice, oat, millet and sorghum. Millet

is a small-grained cereal and it forms the third largest group in world cereal food grains (FAO,

1989). It is highly utilized in many developing countries for food, feed and preparation of

alcoholic beverages. The nutritional advantages of millet are it's high fat content and relatively

high lysine content.

In most developing countries, most children, particularly those in low-income class, are /

weaned on cheap, readily available cereal foods (Altapo et a/., 1995). This can be attributed to 1

several factors including poor nutritional education, decline in household incomes and to a large \ \

extent, the ever increaking cost of commercial formulae. In Nigeria, the usual first weaning food

is pap, referred to as akamu by Igbos, ogi by Yorubas or koko by Hausas. It could be made from

maize p e a mays), millet (Pennisetum an~ericarzum), guinea corn (Sorghum species) or their

combinations (Onofiok and Nnanyelugo, 1998). In Cameroon, millet-based gruels are prepared

for feeding infants.

A major disadvantage of sole cereal gruels is that the starchy nature of these foods makes

them bind so much water, thus yielding a bulky gruel with decreased nutrient content. Increasing

the solids in such gruel to improve nutrient content yields a very thick (viscous) gruel, which

could cause choking in young children (Marero et al., 1988). Besides that, cereals have lower

protein quality compared to legumes. Numerous studies have indicated that cereal grains are the .

major conlributors of energy and protein to the diet in the developing countries and that these

plant foods are low in essential amino acids (EAA) namely lysine and tryptophan (Birthe and

Egum, 1983). Cereal -grain proteins are low in lysine but have adequate amounts of the sulphur

-amino acids (Bressani, 1973).

Legumes are the species of plant family Leguminosae, which are good sources of dietary

protein. They are consumed directly as mature dry seeds, as immature seeds or even as green

pods with the immature seeds enclosed. Examples are cowpea, soyabean, groundnut, pigeon pea

and bambara groundnut. Pigeon pea has been reported to contain high quality protein and is a

good source of amino acids except methionine (Elegbede, 1998). Supplementation of cereals

with legumes has been suggested as one way of improving the protein quality of cereal -based

gruels (Nkama et al., 1995).

1.1 Statement of Research Problem Nutritional disorders among young children in most economically developing countries

have been identified as one of the greatest challenges (Nwanekezi et nl., 2001). Protein -energy

malnutrition have been identified as one of the most important problems in Africa and the high

rate of population growth aggravated this problem. Alteration of the high volume/ high viscosity

characteristic of starch based gruels by lowering its water -binding capacity and eliminating the

anti-nutrient contents could offer a solution to some of the nutritional problems in developing

countries particularly among the low income group.

Based on the afore mentioned problems, this study was designed to:

1. determine the preliminary processing treatment that will positively modify the hnctional

properties of fnillet and pigeon pea grains as well as reduce their antinutrients or detoxify

them.

2. develope weaning food with blends of the modified cereal and legume;

3. evaluate the chemical characteristics and nutritional quality of the formulation.

1.2 Justification for the Study

Millet seed (Penniselzmt nmericnnum) and pigeon pea (Cajanus cajan) are readily

available local materials with very high production levels (FAO, 1995). Therefore, the utilization

( o r these raw materials in different ways would make them beneficial to human beings, thereby,

/ reducing wastage. As population is increasing tremendously, locally available raw materials such

1 as millet and pigeon pea would be used to sustain the population.

Most of the previous studies on weaning food production were based on cowpea,

:joyabcan, maize and rice. These materials have also been competitively used for different

product development by different industries, researchers etc. This could be seen clearly through

the number of researches that have been carried out using these materials and through the

different products gotten from these materials by different industries. ~ c c o r d i n ~ to FA0 (1995),

millet is a cereal whose protein and caloric content are comparable to maize and rice. Till now

people have been eating maize and rice due to the knowledge of what they would derive from the

food in terms of calories and proteins. So far, little have been known about the nutritive value of

millet. Therefore, the knowledge that millet's caloric and protein contents are comparable to that

or maize and rice brings millet to lime light as a good quality protein and caloric food source.

Pigeon pea, on the other hand is a good quality protein food source but it is underutilized due to

ignorance of its nutritional potential (Elegbede, 1998). Oshodi and Ekperigin (1989), reported

that pigeon pea contain a moderate level of crude protein about the same as in cowpea. Thus, the

use of pigeon pea in this study.

In view of the level of use vis-a-vis level of production and local availability of millet and

pigeon pea, the present study dealing on a combination of the two raw materials for the

production of weaning gruel becomes very ,necessary.

LITERATURE REVIEW

2.0 Introduction

Cereals are members of the grass Family, which has edible starchy seeds. They are the

inost important sources of plant food for man and usually provide the bulk of a suitable diet.

Cereals are the major sources of energy and protein in the diets consumed by man. The cereals

cultivated are barley, oats, rye, sorghum, millet, maize, rice and wheat (FAO, 1989).

Climate, soil and custom determine the distribution of the major food grains. Some such

as barley, oats and rye are mainly grown in temperate regions; others such as wheat and maize

are cultivated over a wide range of latitudes. in most tropical climates or in areas where intensive

imgation is practiced, rice is the preferred cereal. In Latin America, and in many areas of sub -

Saharan Africa, maize is the preferred cereal. Wherever ecological conditions are not favourable

to maize, sorghum and millets are grown, as they thrive on relatively poor soils with limited and

uncertain rainfall. Compared with root crops, cereals offer a better source of protein in the diets

of Nigerians whose intake of protein from animal source is low (Okoh, 1998).

Legumes belong to the family Eeguminosae. It is the second most important source of

food and fodder, next only to cereal grains. Legumes are good sources of dietary protein

(Elegbede, 1998). They are cheaper than animal products - meat, fish, poultry and egg -

therefore they are consumed worldwide but in developing countries they serve as a major source

of protein.

2-1 Nutritive Value of Cereal

Mature cereal grains consist mainly of carbohydrates, proteins, lipids, mineral salts and

water. Vitamins are present in small quantities (Kent, 1975). Cereals provide energy from

carbohydrates and fats together with a variable proportion of plant protein, depending on the

species, variety and conditions of cultivation (FAO, 1989). Except for two amino acids, lysine

and tryptophan, most cereals contain essential amino acids required by man as well as vitamins

and minerals.

In many developing countries, cereal is the main source of dietaiy protein where animal

protein is scarce and expensive. The chemical composition of major cereal is shown in Table 1.

The mincrals in the edible kernels of cereals include potassium, magnesium and calcium, mainly

in the form of phosphates and sulphates (FAO, 1989). Some of the phosphorus may be present as'

phytic acid, which may restrict the availability of calcium and iron in the diet.

Table 1. Chemical Conlposition of maior cereal mains ( 100~ edible portion)

Wheat Maize Sorphum Millet Rice

Crude fibre@ 2.0 2.8 2.0 2.3 1 .O

Calories (Kcal) 348 355 3 29 3 63 3 62

Carbohydrates (g) 71 .O 73 .O 70.7 67.0 76.0

Proteins (g) 11.6 9.2 10.4 11.8 7.9

Fat (g) 2.0 4.6 3.1 4.8 2.7

Ash (g) 1.6 1.2 1.6 2.2 1.3

Calcium (mg) 30 26 25 42 33

lron (mg) 3.5 2.7 5.4 11.0 1.8

Riboflavin (mg) 0.10 0.20 0.15 0.21 0.04

Thiaminc (mg) 0.4 1 0.38 0.38 0.38 0.4 1

Niacin (mg) 5.1 3.6 4.3 2.8 4.3

Source: FAO, 1995.

2.2 Nutritive Value of Legume. Legumes are good sources of protein and energy. The chemical and physical properties of

legume seeds vary widely as might be expected from such a broad group of plant produce. Based

on a nitrogen cowersion factor, 6.25, crude protein content of most legume seed varies between

16.0% in bambara groundnut to 35.1% in soybeans (Table 2) (FAO, 1982b). Legume proteins \

are limiting in essential sulphur -bearing amino acids, methionine and cysteine, but are rich in

lysine and tryptophan (FAO, 1982a). Therefore, a combination of cereal and legume proteins

comes close to providing ideal dietary proteins for human beings. Most legumes, except

groundnuts and soybean are low in fat coillent, ranging from 1 to 6%. However, oil seeds have a

range of lipids content from about 18% in soybean to as high as 45% in groundnut.

All legume grains contain substantial amounts of minerals and vitamins -especially those

of the B group, with cowpea, soyabean and bambara groundnut being good source of calcium .

and iron (Elegbede, 1998).

Table 2: Energy and selected nutrients in le~uines (composition per 1OOg

edible portion of dried mature whole seeds). Water Enerev Protctn Fat Carbohvdrates Crude Ash Calcium Iron Thiamine Riboflavin Nicotinic ------

(%) (%) E (a) (d Fibre ($) Lg) (111g) (me) (ma) acid (In$$)

13ambwa 10.1 370 16.0 6.0 65.0 - 3.0 85 4.2 0.18 - -

groundnut

13road bcam 13.8 328 25.0 1.2 56.9 5.1 3.1 104 4.2 0.45 0.19 2.4

~ h ~ c k pcas 1 1.0 362 4 5.6 60.9 2.5 3.1 114 2.2 0.46 0.20 1.2

Cowpeas 11.5 340 22.7 1.6 61.0 4.2 3.2 110 6.2 0.59 0.22 2.3

Groundnuis 7.3 548 23.4 45.3 21.6 2.1 2.4 58 2.2 1.00 0.13 16.8

lack beans 11.2 348 21.0 3.2 61.0 7.6 3.6 134 8.6 0.65 0.13 3.1

IkJncy bcans 12.1 336 20.3 1.2 62.8 4.8 3.6 86 6.9 0.46 0.18 2.0

1an.1 bcans 10.5 346 19.8 1.3 65.4 3.0 90 5.6 0.46 0.2 1 1.4

1'1gco11 pea\ 1 1.5 339 20.4 1.2 63.4 4.4 3.5 103 4.9 0.49 0.2 1 2.2

Soya bcans 10.2 400 35.1 17.7 32.0 4.2 5.0 226 8.5 0.66 0.22 2.2

Source: FAO, 1982

2.3 Anti-nutritional Factors in Legumes

Legumes contain antinutritional factors, which are known to exert deleterious effects in

man and animals, when ingested. These interfere with digestive processes and prevent efficient

utilization 01 the legu~ne proteins. These toxic factors may occur in all parts of the plant, but the

seed is normally the most concentrated source. The levels of deleterious substances in tropical

legumes vary with the spccies of plant, cultivar and post harvest treatments such as drying,

soaking, autoclaving and malting of the seed (Osagie, 1998). Most legume grains are highly

toxic to animals if fed without adequate processing. However, man has through experience learnt

1.0 either avoid some legumes, which produce ill effects or devised a means of eliminating the

toxic components through some kind of processing. The toxic cornponcnts may be divided into

two major categories: [hose proteins, such as lectins and protease inhibitors, which are sensitive

to normal processing temperatures and substances which are stable or resistant to these

temperatures, which include among others polyphenolic compounds (mainly condensed tannins), .

non-protein amino acids and galactomannan gums (Osagie, 1998). More often than not, a single

plant may contain two or more toxic compounds, generally drawn from the two categories,

which adds to the difficulties of detoxification. The antinutritional factors that are considered to

be of significant importance to Nigerians who use plants for both human and animal feeding are

shown in Table 3 .

i. Protease inhibitors: Enzyme inhibitors are widely distributed in plant and animal tissues.

Protease inhibitors are found in cowpea, groundnuts, pigeon pea and liina beans. Tnfact, all

legumes have bcen found to contain trypsin inhibitors to varying degrees, in addition to

chymotlypsin inhibitors. Trypsin and chyrnoptrypsin inhibitors activity in uncooked animal feed

has bcen known to cause diminished growth in rats, chickens and other experimental animals

(Osagie, 1998). Trypsin inhibitors inhibit the trypsin and chymotrypsin in human and leads to the

hypertrophy of pancreas. It reduces protein digestibility, thereby reducing the nutritive value

(Osho, 1989).

From a practical standpoint, trypsin inhibitors do not appear to be a serious problem in

feeds arid foods since they, being proteins, are largely ina~tivatcd by moist heat. Conditions of

heating -time, temperature, moisture content and particle size -influence the rate and extent of

trypsin inhibitor inactivation, lor example, atmospheric steaming at 100°c for 60 minutes

inactivates more than 95% of the trypsin inhibitor activity of raw, defatted bean flakes (Liener,

1975).

ii. Lectins: Lectins are proteins, which are characterized by their unique ability to bind specific

sugars or glycoproteins. This reaction is manifested in vi1r.o by the agglutination (clumping) of

red blood cells from various species of animals and so lectins arc also called haemaggluiinins.

Most lectins are glycoproteins containing 4 to 10% carbohydrates and htive molecular weights

ranging from 100,000 to l5O,OOO (Liener, 1983).

Lectin toxicity is also due to their production of intestinal lesions and histopathological

changes of some organs particularly in the kidney (Ikegwuonu and Bassir, 1977).

I-Iae~nagglutinin activity of extracts from different legumes differs in their potency (Liener,

1975). Detoxification of haemaggh~tinins is usually achieved by the traditional methods of

Table 3: Examples of Antinutritional factors in pi nt foods.

Class Distributions hysiologicral effect

Proteins Prolecase Most legumes, root and tuber Depressed growth, hypertrophy / inhibitors crops hyperplasia, aciilar nodules.

Lectins Amylase Most legumes, root and tuber Interference with starch digestion. inhibitors crops

Glycosides Gynogenic Cassava roots, some legumes, Respiratory failure, gl y cosides sorghum goitre. Oligosaccharides Most legumes Flatulence Saponins Most legumes and tuber crops Affects intestinal permeability.

Phenolics Tamins Most legumes, root crops and Interference with mineral

h i t s . availability. Gossypol Cotton. hterferes with protein digestion

Miscellaneous Phylate Most legumes and root crops. Interference with mineral

availability. Oxalate Vegetables, root and tuber crops. Interference with mincral

availability. Alkaloicls ..- Soine v a n species and kolanuts ~ c ~ ~ s s c d growth.

Soarcc: Liener (1 989). . household cooking and industrial autoclswing or retorting. Complete detoxification could be

achieved by preliminary soaking, prior to cooking.

iii. Y mylase inhibitors: Amylase inhibitors, found in most legumes, inhibit the action of

prmcseatic and salivary amylases. This results in increased amounts of undigested starch in the

faeces and subsequent decrease in the nutritional value of the foodstuffs. Amylase inhibitors are

readily inactivatcd at 100°C. Pre -incubation of the idlibitor with starch resultcd in complete

abolition of its activity (Osngie, 1998).

iv. Goitrogens: Goitrogenic subslances, which cause enlargement of the thyroid gland, have

been found in legumes such as beans and groundnuts, although these are common in edible

plants of the cabbage family. Consumption of inadepately processed or raw soybeans and

groundnuts is responsible for goiter in solnc infa.ilts fcd soymilk and rats fed imp-opcrly

processed groundnuts. The goitrogenic effect was effectively counteracted by iodine

supplementation rather than heat treatment (Liener, 1975).

v. Cyanugenic glycosides: Some legumes contain cyanogenic glycosides from wlich

hydrogen cyanide (HCN) may be released by hydroloysis. Hydrogen cyanide (HCN) is very

toxic at low concentrations to humans and other animals. Hydrolysis occurs rapidly when the

ground meal is cooked in water and most of the liberated hydrogen cyanide (HCN) is lost by

volatilization. Soaking of sceds in water for 24hrs led to appreciable losses of hydrogen cyanide

(HCN), whilc boiling for 3111-s caused drastic reduction. After boiling for 3hrs, innocuous

hydrogen cyanide levels were obtained with seeds previously soaked in water for 24hrs and

divested of testa. Thus, soalung of bean seeds in water and subsequent removal of testa prior to

boiling will decrease hydrogen cyanide in bear1 meals (Okolie and Ugochnkwu, 1989).

vi. Flatulence Factors: One of the major constraints to the human consumption of legumes is

their ability to produce gas in the gastrointestinal tract, which is referred to as flatulence. The

flatus gases produced include carbondioxide, hydrogen and to a lesser extent methane. The

production of tlatus by monogastric animals is due to colonic fermentation of carbohydratcs that

cscape breakdown in the stomach and small intestine. Thcse oligosaccharides; rafinose,

stacl~yosc and verbnscose, wllich are currunon in legunme seeds, are thought to be major

producers of flatdence when these foods are consumed. Animals and inan are not able to digest

such oligcsacclia,ridcs because of the absence of a, - 1, 6 -galactosidase in their intcstind

1nucos9. t

Owing to the absence of enzymes in human i~ltestinal mucosa [hat are capable of

hydrolyzi,~~ this li~ka,ne, the intact oligosacchariclcs accur?xdnte in the lower intestine where thcy

~m!c~-go fc'crmentatio~; by anmxobic bacteria and produce the hove gases. T l~c gases produced / I

are responsible for ch;)ractei-istic features of flatule~lce ~~arrzely nausea, abdominal cramps, \

diarrhea, abdominal rumbling and the social discomfort an,r:o~iated with ejection of rectal gases \

(Davidsoa c/ al., 1975). In both rnls and iiurnans, an induction period of about 4 hours is rcquired

before ilatus gases are passed. This delay corresponds to thc time required for rlze ingested food

to reach the region o r ~ h e inleutine domiilatetl by anaerubic bacicria.

Flatulence factors arc eliminated by propcr heat treatment, knneiltiliion, gcmnination,

protein isolation md dehulling. It is known that the 01igasacchu;dcs are concentrated in the bean

h d or testa, :nd are at a low levcl in the cotylcda~s. 7'h).ls, their removal during dehulling

significantly reduces their level in the beans and their products. Researchers have cancludecl that

thc use of fermented pulse products may reduce the problem of flatulence experienced by mmly

with the ingestion of legumes (Goel and Verma, 1980).

vii. Saponins: Saponins me steroid or triterpenoid glycosides that are characterized by their

bitter or astringent taste, foaming properties and their hemolytic effect on red blood cells. hnong

plants grown for food, the presence of saponins in legumes such as soya and pea is particularly

important. Saponins have been shown to posses both beneficial (cholesterol --lowering) and

deleterious (cytotoxic, permeabilisation of the intestine) properties and to exhibit s w c b e

dcpendcnt biological activities (Price et al., 1987).

Alkaline washing or dry scouring and abrasive dehulling have been suggested as

techniques of reducing saponins in legumes. In general saponins are not destroyed during

cooking or processing.

viii. Tannins: l'annins are phenolic compounds with molecular weight. greater than about 500.

There me the hydrolysable tannins, which could be hydrolysed into a mixture of carbohydrate

and phenols, and the condensed tannins, which are complex ilavonoid polymers.

Polyphenols (as condensed tannins) are predominantly located in the pericarp and /or

testa, particularly of pigmented cultivars of legumes and millets (Chavan et al., 1979, Deshpande I

er al., 1982). Tannins decrease protein quality by decreasing digestibility md palatability.

Tannins, which are also known as non -specific inhibitors of enzymes, may reduce protein

quality by directly complexing with food proteins (Davis and Hoseney, 1979, Anon, 1983). Elias

et al., (1979), reported that tannin concentnition was high in coloured sced coats ranging fiom 38

-43mg/g and low in white coated beans (1.3mdg). Bressmi md Elias (1980), observed a higher I

I

protein quality for white beans as compared to pigmented cultivars of dry beans. Tannins

concentration ranged from 0 - 0.7% for cowpea, 0 - 0.2% for pigcon pen, with essentially no

tannins found in chickpea or mung bean. Hoff and Singletons (1977), reported that tannins \ interfere with digestion process by inactivating enzymes in the alimentary canal and increasing \

fecal nitrogen. Their protein coinplexir~g ability renders them astringent in contact with mucous

membranes of the oval cavity and thereby affecting palatability.

Diets high in tannins have been shown to cause interference with paricrealic digestion,

irritation of the intestinal tract and thus give rise to ineihionine deficiency by requiring active

metliyl groups in the detoxification process. High tannins content can also reduce the

bioavailability of iron (FAO, 1989). Other nutritional effects, which have been attributed to

tannins, include damage to the intestinal tract, toxicity of tannins absorbed fiom the gut and

interference with the absorption of iron and a possible carcinogenic effect (Butler, 1989).

People have learnt how to use their own cultivars as sources of digestible food. $ome of

these methods include germination, soaking in water for various length of time and discarding

the soak water, adding alkali such as wood ash, palm bunch as11 (containing potassium

carbonate) or mineral limestone or trona, h i t juices or tamarind pulp, when preparing food.

ix. Phytate: Phytic acid, a hexaphosphate derivative of inositol is an important storage form of

phosphorus in plants. It is insoluble and cannot be absorbed in the human intestines. Phytic acid

has 12 replaceable hydrogen atoms with which it could form insoluble salts with metals such as

calcium, iron, zinc and magnesium. The formation of these insoluble salts renders the metals

unavailable for absorption into the body.

2.4 Antinutritional Factors in Cereals

A major factor limiting the wider food use of many tropical plants is the r~biquitous

occurrence in them of a diverse range of natural compounds capable of precipitating deleterious

effects in man and animals. Manifestations of toxicity range from scvcre reduction in food intake

and nutrient utilization to profound neurological effects and even death. Compounds, which act

to reduce nutrient utilization andlor food intake, are often referred to as anti-nutritional factors

(ANF). l ' l ~ antinutritional factors associated with cereals include:

i. Phytate: Phytate represents a complek class of naturally occurring phosphorous compounds

that can significantly influence the functional and nutritional properties of hods. Phytic acid,

nq~oinositol -1,2,3,4,5,6 -hexakis (dihydrogen phosphate), is the main phospl~orus store in

mature seeds. Phytic acid has a strong binding capacity, readily forming complexes with

multivalent cations and proteins. Most of the phytate -metal cornplexcs are iixolublc at

physiological pH. I-1encd phytate binding renders several minerals biologically unavailable to

animals and l~umans.

Pllyta~e is found in germ of corn and in the bran and germ of other cereal grains because

ii fhctions as a storage form of high-energy phosphorus for the geminating seed (Soulhgate,

2993). Doherty et a/. (1982), analysed several varieties of sorghum and found that in ihc wholc

gain, phytin phosphorus ranged from 170 to 380mg per 100g; over 85% of thc total phosphorus

in the whole grain was bound as phytin phosphorus. Sankaro Rao et al. (1983), reported that the

values for phytin phosphorus in pearl millet varied from 172mg per 1 OOg to 327mg per 100g. I

Phytate reduces the bioavailability of calcium, copper, iron, magnesium, and zinc, and

may interfere with protein metabolism. It decreases the utilization of protein subjected to

proteolytic digestion and in most cases, protein serves as a bridge between phytate and mineral

interaction (Deshpande et al., 1982). Calcium forms a complex with phytate making it

unavailable for absorption. Phytate also reduces the availability of iron in the body foiming an

insoluble bond in a non -ferric foim, a foim that is more readily absorbed and utilized by the

human body (O'Dell, 1983). Reduction in phytate level in wheat bran improved zinc

bioavailability (Morris and Elis, 1980). Sankaro Rao et al., (1983), observed that malting of the

grain significantly reduced the phytin phosphorus content of both pearl and finger millets. This

decrease was accompanied by significant increases in ionizable iron and soluble zinc, indicating

improve bioavailability of these two elements.

It is therefore important that during processing that the toxic substances be reduced to

levels that pose no threat to health. Processing method may include dehulling, cooking,

germinating, soaking, fermentation among others (Walker and Kocher, 1982). During

germination, phytase activity increases while phytate content decreases (Southgate, 1993).

ii. Tannins: Tannins are polyphenols, which are widely distributed in plants. Some sorghum

varieties contain tannins that are polyphenols. These impart reddish to brownish colour to

sorghum grain and behave as nutritionhl inhibitors because thcy combine with proteins and make

them indigestible and unavailable to the body. Therefore sorghum grains should be properly

processed before being used as food, especially fbr humans because tannins in sorghum are

associated with poor protein utilization. Some varieties of sorghum containing high tannins in the

grains were found to be bird resistant (Burns, 1971, Tipton et al., 1970). Tannins are the most

abundant phenolic coinpounds in brown bird -resistant sorghum. During maturation, the brown-

sorghum develops astringence, which imparts resistance against bird and grain modd attack

(FAO, 1995).

Tannins, while coderring the agronomic advantage of bird resistance, adversely affect

the grains nutritional quality (Salunke et al., 1982). Among millets, finger millet was reported to

contain high amounts of tannins ranging fi-om 0.04 to 3.47% (FAO, 1995). Growth retardation

was observed in chicks fed high -tannin sorghums. Tannins in the grains impart an astringent

taste, which affects palatability; reduce food intake and consequently body growth. Tannins bind

to both exogenous and endogenous proteins including em rnes of the digestive tract, affecting

the utilization of proteins (Asquith and Butler, 1986).

Tannins and associated polyphenols are concentrated in the testa or seed coal and can be

removed by milling. Germination was also found to decrease tannin content in sorghum

(Osuntogun et ul.; 1989) and finger millet (Udayasekhara Rao and Deosthale, 1988). However,

the tannin content of the germinated sorghum rose again significantly upon drying.

iii. Digestive Enzyme Inhibitors: Inhibitors of amylases and proteases have been identified

in sorghum and some millet. It was reported that sorghum had the highest inhibitory activity

against human, bovine and porcine amylases, foxtail millet did not inhibit human pancreatic

amylase, while extracts iiom pearl and finger millets inhibited all a - amylases tested

(Chandrasekher ef al., 1981). Finger millets extracts were found to have highest activity against

bovine trypsin and chymotrypsin. Extracts from sorghum and millets inhibited the proteolytic

cnzyme of both human and bovine pancreatic preparations (FAO, 1995).

Finger millet was found to have inhibitory property against salivary and pancreatic

amylases. The inhibitors iiom sorghum and foxtail millets were more therrnolabile than those

from finger and pearl millet.

iv. Cyanogenic glycosides: Cyanogenic glycosides are a group of 0 -glycosides formed from

decarboxylated amino acids. The cyano group arises from the a-- carbon atom and the amino

group. The occurrence of cyanogenic glycosides in crop plants such as cereals is well known.

According to Doggett (1988), cyanogehic glycoside, dhurrin occurs in most sorghum varieties,

although the quality depends on the variety and environmental conditions. The use of sorghum as

human food or for livestock feed is seriously limited by the presence of dhurrin in its seeds,

shoots and roots. The level of this poisonous natural product depends among other factors, on the

sorghum, sprouting and component part of the sorghum sprouts (Ikediobi et ul., 1988).

Evidently, dhut-rin is highly toxic owing to its ability to produce hydrogen cyanide when

hydrolysed.

Some local foods and beverages produced from sprouted sorghum grains contained

negligible or undetectable levels of cyanide. Apparently, sprouting and rubbing off of roots and

shoots coupled with heat or hot water treatment during processing has been used in detoxifying

sorghum -based food and beverage products (Osagie, 1998).

v. Goitrogens: Iodine is an essential micronutrient for all animal species, and iodine deficiency

is among the most widely prevalent nutritional problems in many developing countries

(DeMayer et a/., 1979). Though environmental iodine deficiency is a prerequisite to goiter

formation, the incidence of goitre in animals and humans with normal dietary intake of iodine ,

suggests there are other factors in the aetiology of simple goitre. A large number of foodstuEs

possess antithyroid agents, collectively designated as goitrogens.

Pearl millet is a staple food implicated in the aetiology of goitre. Consumption of pearl

millect is considered one of the factors responsible for the high incidence of goitre in rural

populations.

Iodine supplementation did not alleviate the giotrogenic effect of pearl millet. Tempering

the grain to 26% moisture overnight prior to milling resulted in flour with no goitrogenic activity

(Klopfenstein et al., 1 99 1).

2.5 Detoxification Treatments Different technologies and treatments are employed in the detoxification of grains for

manufacturing nutritious commercial weaning foods. These treatments include:

Dehulling and/ or Peeling: Dehulling and/ or peeling remove unwanted parts of raw materials. ,

Dehulling and decrotication are used synonymously in cereal and legume processing for removal

of testa (Enwere, 1998). Dehulling or peeling significantly reduces the levels of poisonous

phytotoxins such as cyanogenic glycosides in tuber crops example, cassava. Alkaloids, tannins

and other polyphenols in pigmented seeds are significantly reduced by dehulling or decortication

treatment (Nout and Ngoddy, 1997). Removal of aleurone layer of the seed bran eliminates

significant levels of phytates, which bind calcium and other minerals. In infant formulas,

decortication reduces the total level of indigestible fibres so that infants are able to handle \ legumes earlier in their diet (Nout and Ngoddy, 1997).

Washing and Soaking: Washing and soaking involve washing and extended steeping in an

excess volume of cold or warm water. Soaking induces the leaching out of water-soluble anti-

nutritional factors. Glycosides, alkaloids, phytates, oligosaccl~arides and tannins are all

significa~itly reduced. Although leaching also loses water-soluble micronutrients, extended I

soaking has the net effect of enhancing the protein solubility index and the availability of

limiting amino acid of edible grains mout and Ngoddy, 1997).

Sprouting or Germination or Malting: Germination is a natural process in which dormant

but viable seeds are induced to start growing into seedlings. During germination, hormones are

produced and enzymes are mobilized to convert stored foods such as insoluble carbohydrates and

proteins to soluble components that are more easily assimilated.

Germination is known to increase the vitamin C, E and B -complex contents of seeds

(FAO, 1995). Rootlets and sprouts of germinated seeds contain very large amount of cyanogenic

gly-coside, which on hydrolysis produces a potent toxin known as hydrocyanic acid (HCN) and

cyanide (Panasiuk and Bills, 1984). It was shown that the removal of the shoots and roots

reduces the hydrocyanic acid content (FAO, 1995). Germination decreases the antinutrient

content of seeds such as tannins, phytates, lectins and oligosaccharides. Germination of grain is

also reported to change the amino acid composition, convert starch into sugars and improve the

availability of fat, vitamins and minerals (FAO, 1995).

Fermentation: Fermentation is a process of anaerobic or partially anaerobic oxidation of

carbohydrate especially sugars, that have been subjected to the action of microorganisms

(bacteria, filamentous hngi or yeasts) or enzymes to produce desirable biochemical changes

(Enwere, 1998, Ency. MCB, 1992). The microorganisms may be the microflora indigenously

present on vegetable or animal products that serve as the substrates for fermentation or they may

be added starter cultures. During fermentation, all microorganisms (bacteria, yeasts and moulds)

take parL in catabolic processes, which alter the organic components of the food to obtain energy

for their growth. Bacteria are mainly* responsible for the fermentation of cereal and animal

products. The major types of bacteria important in cereal fermentation are those that produce

lactic acid from available sugar.

Fermentation has been reported to enhance nutritional value, texture, shelf life, aroma

and taste of food products (Onuoha and Obizoba, 2001). Fermentation of grains either as

porridge or as slurries of the flour has been reported to significantly improve the protein

digestibility of grain (Mensah et al., 1990). Protein solubility and the availability is enhanced in

some cases by as much as 50%. The micronutrient availability is also enhanced because of

significant reductions in phytates. Tannins are reduced by as much as 50% and oligosaccharides

by as much as 90% in some reported cases (Lorri, 1993). Fermentation to below pH 4 has been

shown to inhibit the proliferation of diarrhea -causing pathogens (Mensah et al., 1990).

Fermented foods have been reported to be used for treating diarrhoea and measles in some parts

of the country.

Dry -roasting or toasting: This is a thermal treatment at high temperature, which can be

carried out in simple low -cost mechanical equipment. Problems associated with a high

propensity of wet starches to stick and burn on equipment suifaces during toasting, can be

contained by carehl manipulation of the moisture content and particle size. Because toasting is a

high temperature thermal treatment, it reduces the level of protease inhibitors and lectins. It also

reduces the level of volatile glycosides that may be present. Because of significant degrees of

dextrinization of starches during high -temperature dry -toasting, it has a diminishing effect on

porridge viscosity. However, this is counteracted to some extent by the increased swelling

capacity of cooked, gelatinized starch.

As a conventional thermal process, dry toasting has a severe adverse effect on protein

solubility as weU as the availability of both limiting amino acids and vitamins.

Extrusion Cooking: Extrusion cooking is a high-temperature short-time (HTST) thermal

process, which cooks, dries and restructures the product in one integrated operation. Significant

levels of drying can result from expansion in the product, which occurs at the extruder die.

Extrusion -cooked -products exhibit quality attributes that are generally superior to those heated

by other means as regards protein solubility and fhctionality, the availability of limiting amino

acids and residual levels of anti-nutritional factors achieved (Nout and Ngoddy, 1 997).

2.6 Use of Cereals and Legumes as Weaning Foods . Cereals are the most widely consumed crops globally. In Nigeria, cereals serve as the

major sources of enerby and protein in the diets of people. Unfortunately, the nutritional quality

of most cereal protein is poor because they contain less of the essential amino acids, particularly 1

fysine needed for growth and maintenance (Okoh et al., 1985).

On the other hwd, legumes are good sources of dietary protein, and are rich in lysine and \ \

\

tryptophan (FAO, 1982a). Therefore, the supplementation of cereal protein with legume protein

would provide an adequate amount of lysine for growth and maintenance. Nkama and Mallesli

(1998), reported increased lysine in a millet -cowpea and rice- millet -cowpea blends. It was

also reported that mixing of two or more

quality than any of the indivi:.!ual sources.

According to Okcke and Obizoba,

sources with different first limiting amino

sources of dietary protein, results in a better protein

(1986), better results are achieved by mixing protein

acids. Maximizing the use of locally available grains 1

and use of cereal--legume combinations to produce high-energy protein formulation have been

suggested as possible solutions to nutritional problenl (Ossai and Maloino, 1988, Nkama and .

Malleshi, 1998).

2.7 Production, Utilizatiou and Nutritive Composition of Millet

Miilet Production: The grains most generally recognized as millets belong to two varieties of

the grass family, the Chlorideae and Paniceae. The tribe Chlorideae includes Eleusine coracana

as the species of economic importance. This plant is variously called Afiican ragi or Finger

millet and extensively grown in India for human food. The tribe Paniceae includes several

species grown for food and feed in various parts of the world. Penniseturn americanunz (Pearl

millet) is extensively cultivated in Egypt and tropical Asia as cereal food. Pearl millet is the sixth

most important of all the world cereals.

Thc major producers of millets in 1990 were India, 11 500 (39%), China, 4401(15%),

Nigeria, 4000 (13%) and the Soviet Union, 3647 (12%) (FAO, 1991). Table 4. The major

producers of millet in 2001 were India, 9505 (33%), Nigeria, GlO5(21%) and China, 2446, (8%)

(FAO, 2002). Table 5.

Table 4: lead in^ Millet Producers, 1990

gountrv Production (IO~MT) % of Total

India 1 1500 38.6

China 440 1 14.8

Nigeria 4000

USSR 3647

Niger 1133

World .-. 29817

Source: FAO, 1991.

Table 5: Leadinp Millet Producers, 2001.

Country -. Production ( ~ o ~ M T ) % of Total India 9505 32.5

Nigeria 6105 20.9

China 24 46 8.4

Niger 2414 8.3

Burkina Faso 957 3.3 . World 29207

Source: FAO, 2002.

Millet Utilization

Of the 30 million tonnes of millet produced in the world, about 90% is utilized in

developing countries and only a tiny volume is used in the developed countries outside the

former Soviet Union (FAO, 1995). It was estinlaied that a total of 20 million tonnes are

consumed as food, the best being equally divided between feed and other uses such as seed, the

preparation of alcoholic beverages and waste (FAO, 1995).

Pearl millet suffers less from discases than sorghum, 111aize or other grains. It has more

oil than maize and is a "high-energy" cercal. It is used mainly as whole, cracked or ground ilour,

dough or a grain like rice. Millet seeds are cooked in the sane way as rice, or ground or pounded

into porridge, couscous, cakes or unliavened bread. Millets are made into unfernlented breads

(roti), fern~ented hods (Kisra and gulletes), thin and thick porridges (toh), steam -cooked dishes,

non -alcoholic beverages and snacks (BOSTID, 1996). ,

A non -alcoholic clrink is made tiom the flour. In Nigeria, pearl millet is fermented, like

maize and sorghum, to produce 'Ogi' - n traditional weaning food. In West Afica, pearl millet is \

snanalted and used for n&ng beer. Whole plants can be used as cash crops and green illamre. \

Nutritive Composition of Millet: The chy grain of pearl millet is usually made up of about

70% carbohydrate, which consists mostly of starch. The protein content (1 1.8%) is comparable

to that of wheat and maize (Table 1). Pearl millet contains high levels of fat and crude fibre. One

of the characteristic features of grain composition of millets is their high ash content. They are

aim relatively rich in iron and phosphorus. The whole grain is sul important source of B-

Complex vitamins, which are mainly concentrxtted in the outer 1)rm layers of the grain. Pearl

millct contains a variation of 9 - 13% protcin. The essential amino acid profile shows more

lysinc, threonine, mellzionine and cystine in pearl millet than in sorghum. It also has high .

tryptophan content (Table 6). It contains higher protein and energy levels than maize or sorghum.

Total dietsuy fibre in pearl millet is higher than that in sorghum, wheat and rice. High fibre

content and poor digestibility of nutrients are other characteristic features of millet grain, which

severely influence their consumer acceptability. Mincral content of peal millet is of a wide

variation but was found to be poor in available zinc, iron and manganese when compared with

sorghum. Malting enhanced the ionizable iron content and increased the soluble zinc content of

pearl millet, indicating an improvement in in vitro availability of these two elements (Smkaro

Rao et af., 1983).

2.8 Production, Utilization and Nutritive Composition of Pigeon Pea.

Pigeon Pea Production: The pigeon pea (Cajcmus cajan L.) belongs to the legunlinoseae

family of flowering plants.

Table 6: Essential amino acid composition (mdg) and chemical score of naillet

and sorr;lbum proteins, -- Amino acids Pearl millet Sowhum

Lysine 214 126

Isoi eucinr; 256 245

Leucine 598 532

Methionine 154 8 7

C yslirie 148 94

Phenyla1,mine 30 1 306

Tyrosinc 203 167

'i3rconine 24 1 180

Typ to phan 122 63

Vali ne 345 3 13

It is an erect wljody short pere~ulial shrub, which grows in semi -arid and sub -.humid

tropics, with deep exknsive root systcm. 1.t is a little known crop, which at present in Nigeria, is

cultivated mostly in the Northern States. The Major producers of peas are Canada, 21 96(21%),

France, l680(16%), China, 1 100(1 I %) and Rusian fed, 1000(10%) (FAO, 2002) (Table 7).

Major producers of pulses are Canada, 3559(7%), Nigeria, 2200(4%), Mexico, 141 1(3%) and

USA, 1228(2%) (FAO, 2002). (Table 8).

Pigeon pea Utilization: Pigeon pea is consumed either alone or in combination with starchy

staples (cereals, tubers and roots), after hours of boiling to destroy toxins inherent in most

legumes and to soften the hard seed coat (Obizoba, 1983). 'I'he peas when grccn can bc

consumed as vegetables and as dry beans when dry and mature. In ~ i ~ e i i a , the dry matter seeds

are cooked whole until tender.

Table 7: Leading Pea Producers, 2001.

Country Production (IO~MT) O h of Total

Canada 2196 20.89

France 1680 15.98

China 1100 10.46

Rusian fed 1000 9.5 1

India 700 6.66

Ethiopia 147 1.39

World 10.5 12

Source: FAO, 2002.

Adults and older children consume the foods. Infants and weaning children, whose

digestive capacities are limited by age, cannot utilize such foods effectively. It has been shown

that flour rather than whole grains is a better source of good quality protein foods for infants and

small children, as judged by growth, liver weight, liver nitrogen (N) and plasma proteins

(Obizoba, 1983).

Table 8: Leadine Pulses Producers, 2001.

Country Production ( ~ o ~ M T ) % of Total

Canada 3559 6.79

Nigeria 2200 4.20

Mexico 141 1 2.70

[JSA 1228 2.34

Ethiopia 1050 2.00

World 52385

Source: FAO, 2002.

Matured seeds are boiled and mashed with cooked potatoes or boiled bananas and eaten

with greens, or boiled and mixed with maize. They are also fried with meat and vegetable to

make stew. In India, the dried seeds are prepared as flour or split as dhal, which is added to soups

or eaten with rice. Often used as food for children. The sprouted seed is popular as a lightly

cooked vegetable.

The immature or green seeds and the pods are used as a vegetable or canned. In Malawi,

the young seeds are removed from the pods and eaten as a snack between meals. l'he tender

leaves are occasionally used as a pot-herb. It is also used as animal feed.

Pigeon pea Nutritive Composition: The fresh immature unripe pigeon pea comprise 45% of

the weight of the whole pod. In this form, they contain about 67.4% water, 7.0% protein, 0.6%

fat, 20.2% carbohydrate, 3.5% crude fibre' and 1.3% ash (Bressani, 1975). When the seeds are

mature and dry, they contain 11.5% water, 20.4% protein, 1.2% fat, 63.4% carbohydrate, 4.4%

crude fibre and 3.8% ash (Table 2) (FAO, 1982b).

The quality of the protein as in other foods is determined by the content of the amino

acids, especially the essential ones. Tryptophan levels are low and may be improved by the

addition of cereal proteins or the pure amino acids. It was reported that the supplementation of

pigeon pea protein with DL - Methionine and DL - Tryptophan improved weight gain and

protein efficiency ratio of diet (Bressani, 1975). The pigeon pea protein is highly comparable

with soybean protein in its content of essential amino acids other than methionine, as presented

in the amino acid profile (Table 9) (Apata and Ologhobo, 1994). Thus, pigeon pea has high

quality protein and is a good source of amino acids except methionine.

Potassium and magnesium are the predominant minerals of pigeon pea and it's mineral

content compares favourably with that of soybean (Osagie, 1998). The seeds are reported to

contain trypsin and chymotrypsin inhibitors but these are destroyed by thorough cooking (Rachie

and Silvestre, 1977).

2.9 Compositions and Nutritional Requirement of Weaning Foods

The combination of breast milk and complementary food should provide all the nutrients

required for growth and development. Mitzner et al., (1984), illustrated in a food square the

concept of an adequate complementary food as shown in fig. 1.

In the food square, breast milk appears in the center of the square to indicate that, as a

complete food, it is the principal part of the diet. Breast feeding is depicted as concentric circles

of decreasing sizes to indicate that its contribution to the total dietary intake decreases with age.

Even when the daily intake of human milk is small compared to weaning foods, it is a valuable

source of high quality protein, energy and other nutrients.

Source: Mitzner et al., 1984.

Table 9: Amino acid composition (ell6eN) of the protein of raw lepume seeds.

Bambara Kidney Lima Pigeon Jack

Amino acids Groundnut Bean Bean Pea Bean

Essential

Isoleucine 3.98 6.0 1 5.26 3.96 5.12

Leucine 7.59 8.38 8.37 8.04 9.07

Lysine 6.83 6.7 1 6.90 6.82. 6.56

Methionine 1.29 1.32 1.24 1.37 1.47

Cystine 1.40 1.18 0.98 1.24 0.89

Tyrosine 3.37 4.46 4.02 3.73 4.15

Phenylalanine 5.25 5.26 6.46 8.62 5.97

Threonine 3.78 5.07 4.45 4.12 4.37

'Tryptophan 1 .19 1.12 1.03 0.86 1.02

Valine 4.81 5.94 6.10 4.70 5.85

Non-Essential

Aspartic -acid 1 1.43 1 1.90 13.17 10.35 13.50

Glutamic-acid 16.60 14.26 15.90 17.32 14.81

Alanine 4.74 4.61 6.20 6.17 4.95

Glycine 3.82 4.26 5.13 4.15 4.63

Histidine 2.94 3.20 " 3.01 3.08 4.56

Proline 4.24 3.85 5.40 3.89 4.10

Arginine 7.16 5.56 6.24 6.48 6.90

Serine 5.26 4.39 6.88 4.58 7.01

Source: Apata and Ologhobo, 1994. \

The nutritional requirements of infants include energy, protein and fluid. Nutritional \

requirements of infants and young children vary from 2200KJ (under lyr) to 6500KJ (1 -5yrs)

(Garnan and Sherrington, 1990). Young children have high-energy needs in relation to their

small size since they are active and growing fast. The energy requirement of an infant is about

700kcal/day for an infant of three of six months old, the age range when weaning usually begin

in most developing countries. As thc infant grows to about 241nonths of age, breast milk is no

longer its principal food because the requirement would have risen to 1350kcal/day.

The absolute protein requirement of an infant increases with age from about 26glday for a

three to five years old child. The vitamins and minerals intakc arc given according to ages. 'l'hc

intakes of energy, protein and fluids recommended by the joint Food and Agricultural

Organization (FA())/ World Health Organization (WHO)/ United Nations University (UNU)

Expert Committee on protein -energy requirements are given in Table 10.

Table 10: Daily Averape Energy and Fluid Requirements and Safe level of

Protein Intake for Infants and Children three months to five years, Sexes

combined (FAOIWHOIUNU, 1983).

AGE

MONTHS YEARS

3 - 6 6 - 9 9-12 1 - 2 2 - 3 3 - 5

Approx. Weight (kg) 7 8.5 9.5 1 1 13.5 16.5

Energy Requirement 700 810 950 1150 1350 1550

(Kcall day)

Safe levels of protein 17 19 19 18.5 22 26

(g/day

Water Requirement 900-1100 1050-1250 1 150-1300 1250-1400 - 1650-1 800

Mitzner et al., (1984)

2.10 Protein Avgilability in Weaning Food

In many developing countries, where animal protein is scarce and expensive for the

average income earners, the main dietary source of protein is of plant origin (FAO, 1989). Cereal

proteins accounts for the major portion of dietary proteins consumed in developing countries, \ k

\ therefore the availability of cereal proteins for absorption and meeting individua1 protein needs is

of significant importance. Cereal protein is usually of good nutritive value, especially when

cereals are consumed with legumes in the same meal.

The quality of a protein is primarily a fimctioil of its essential amino acid composition.

Fluctuation in the protein content of the grains are generally accompanies by changes in the

amino acid composition of protein (FAO, 1995). Most cereals, including sorghum, maize and

millets are limiting in essential amino acids like lysine, threonine and tryptophan and this makes w

their protein quality poorer compared with animal irotein (Lorri, 1993).

Apart from a favourable essential amino acid profile, easy digestibility is an important

attribute of a good quality protein (FAO, 1995). The protein digestibility of cereal is generally '

lower than the digestibility of animal proteins, partly due to the presence of fibres and tannins,

which bind to protein, thus making it indigestible (Graham et nl., 1980). Reduction of the

disulphide bonds increased protein digestibility of sorghum (Hamaker et al., 1986). Cereals like

sorghum have been shown to have low protein digestibility because of increased levels of

disulplude cross linkages in sorghum prolamin proteins (Hamaker et nl., 1986).

In a few studies on humans, mainly children, the apparent protein digestibility was

reported to be 81% for refined wheat flour, 67 - 75% for dehulled rice, 73% for whole maize

kernel and 46 - 55% for whole sorghum grain (Lorri, 1993). It was found that the protein

digestibility of legumes ranges from a low 72% for black beans to a medium 89% for chick peas

and to a high 90 - 98% for soybean products (FAO, 1991). Lactic acid fermentation of whole

grain flour increased the protein digestibility of sorghum to 73% (Lorri, 1993).

2.1 1 Dietary Bulk in Weaning Foods

The concept of dietary bulk refers to the factors in the food that make it diflicult for an

individual to consume the food in sufficient amounts to cover hislher energy and nutrient

requirements (Lorri, 1993). The energylnutrient density and the consistency of the diet are the

two characteristics identified as the dietary bulk factors (Svanberg, 1987). According to

Ljungqvist et al., (1981), the amount of food that children can eat per meal is influenced by the

amount swallowed and the capacity of their stomachs, both of which increase with age. On the

other hand, the amount of food that a child actually eats depends on hislher appetite, on the

consistency and palatability of the food, as well as the patience of the person feeding the child.

The energy density of most typical weaning food (gruel) is normally very small. The thin

porridge contains around 5 -10% dry matter, which would give 0.2 - 0.4 kcallg of prepared food.

If cooking oil or sugar or groundnuts are added the energy density will be increased (Ljungqvist

et al., 1981).

As the children grow older, approaching one year of age, the mothers may prepare thicker

gruels. A dry matter content of 20% is normally the upper limit, as the gruel becomes difficult to

stir beyond this concentration. The use of flour from germinated cereal (power flour) for 9

reducing dietary bulk problems has been success~lly demonstrated in Tanzania (Mosha and

Svanberg, 1990), India (Gopaldas ef a!., 1988) and Chile (Alvina el a!., 1990). 'The tcchnique of

using power flour results in a breakdown of the starch gel network in the porridge prepared from '

ungerminated cereal flours. Thus, the addition of germinated flour may allow the amount of flour

of porridge to be increased several times without thickening the consistency.

Fermentation and germination processes may thus be used together to reduce the dietary

bulk of cereal - based gruels.

2.12 Weaning Foods and Practices in West Africa

The purpose of a weaning food is to add to or complement mother's milk. For this reason

the weaning process is sometimes referred to as "Complementary feeding". Compared to

supplementary feeding, the term "Complementary" emphasizes the nutritional objectives of

prolonging breast feeding and adding to or complementing the nutrients provided by breast milk

rather than replacing the nutrients as the child grows. In Nigeria the usual first weaning food is a

thin gruel, called pap and is introduced at three to six months of age. The baby is fed on demand

with a spoon or a cup. After the successfbl introduction of cereal gruel, other staple foods in the

family menu are given to the child. These foods include yam (Dioscorea spp), rice (Oryza

saliva), garri (fermented cassava grits) and cocoyam (Xanlhosomcr s~tli$oli~im) that may be

eaten with sauce or soup. These foods are usually mashed, thinned or pre-chewed. Legumes are

rarely used for weaning and are introduced much later (after six months of age), probably

because of the problem of indigestibility, flatulence and diarrhoea associated with their use

(Onofiok and Nnanyelugo, 1998).

The most common practice in Ghana is that mothers starts weaning by the third month of

life. The main weaning food for infants up to six months of age is a traditional fermented maize

porridge (koko). From six months onwards, the infants are given the family diet with

complementary breast-feeding. The family foods on which the infants are weaned include dishes

made from cereal, starchy tubers, legumes and vegetables.

In Sierra Leone, ogi prepared from maize or sorghum (couscous ogi) is a popular

weaning food. Other West African countries have similar weaning practices.

2.13 Previous Studies on Weaning Food Formulation

Weaning foods have been formulated from different cereals, legumes, tubers and roots. It

was found that pigeon pea protein was improved by the addition of rice (20%) to the pigeon pea

(80%) and that the pattern of amino acids produced by the mixture (80:20) were utilized as those

firmished by casein (Obizoba, 1983). Similar investigation with sprouted and fermented

sorghum, bambara groundnut and sweet potato flours by Chima (1998) showed that the nutritive

values, nutrient density and low viscosity of such foods can be improved by sprouting and

fermentation. Obizoba (1985), found that rats fed with combinations of dehulled brown bean

(DBB) or dehulled white bean (DWB) with corn flour had increased food intake, weight gain,

Nitrogen intake, digested and retained Nitrogen and liver Nitrogen compared to those of the

casein control group. It was also found that the combination of rice and bambara groundnut

(80:20) produced increases in food and Nitrogen intakes, weight gain, digested and retained N,

Net Protein Utilization (NPU) and biological value (BV) higher than for those of other diets

which include maize, sorghum, brown bean, white bean, cowpea and pigeon pea (Obizoba,

1986). Okeke and Obizoba, (1986), found that starchy foods when blended with legumes at low

levels produced good quality protein comparable to casein. In the study, carried out on sorghum

and corn by Obizoba (1988), it was found that processes of malting and wet milling improved

the nutritional value of proteins of sorghum and corn by increasing the total protein content and

quality.

Obizoba (1990) noted that combinations of two varieties of germinated cowpea (Akidi

and Oraludi), bambara gtoundnut, pigeon pea with sprouted yellow corn showed that the Akidi

and bambara groundnut blends showed nutritional superiority over other blends as judged by

nitrogen balance and mineral utilization Ogundide (1991) found that weaning food formulated

from composite flour containing 25% cowpea and 75% maize flour is similar in acceptability to

Nutrend, which is a commercial product. The best reconstituting conditions are achieved when 5

times of water to solid is used. In the study, on the effect of sprouting on nutritive qualities of

millet and cowpea flours and their blends, Salami (1991) concluded that water absorption and

swelling of flours decrease with sprouting time. Sprouting increased activity of hydrolytic

enzymes and nutrients except fat and starch.

Corn protein supplemented with breadfruit as low cost vegetable protein or with crayfish

as low cost animal protein would provide protein of good quality that has efficient utilization and 8

high complementary mixture (Chukwuka, 1985). Ebiogwu (1992), found that weaning food

formulated raw before wet milling, steaming, drying and dry-milling at 75% maize: 25% cowpea

was the most acceptable and compared favourably with the commercial weaning foods available '

in our society. In the study carried out by Emenike (1995) on pigeon pea flour and corn flour

blends, it was observed that the legume -cereal 70:30, 60:40, 50:50 ratios gave product that have

adequate nutrients, chemically the best in terms of protein content and organoleptically accepted.

Fermented cereal-legume -tuber blends was found to have better colour, odour,

consistency, more acceptability and leads to more preservable product in terms of its shelf life

(Onah, 1999). Ogbu (1999) concluded that fermentation of cereal -legume blend complementary

foods could lead to the reduction of flatulence factors and also that the pH and viscosity of

fermented flours are lower than that of unfermented flours. Nnam (2001) found that blends from

sprouted sorghum, bambara groundnuts and fermented sweet potatoes (52:46:2) ratio had better

nutritional attributes than their unprocessed counterparts and the traditional sorghum

complementary food. In the study on the formulation of weaning foods from millet, cowpea and

groundnut mixtures (70:20:10) and millet, cowpea, groundnut and millet malt mixtures

(65:20:10:5). It was found that the addition of legumes improved the nutrient content of the

weaning food, while addition of malt reduced the hot paste viscosity, making the weaning foods

nutrients dense. (Nkama et al., 200 1).

CHAPTER THREE

MATERIALS AND METHODS

3.1 Preparation of raw material

Preparation of Seeds:

Seeds of pigeon pea (Cajanus cajan) and millet (Pennisetum americanum) were obtained

from dealers in a local market in Nsukka, Enugu State. Seeds were cleaned by winnowing. hand

sorting and floatation. A 400g weight of seeds was used for each germination and fermentation

studies in the preliminary work.

Germination Procedure:

Whole seeds were weighed, washed and soaked separately in a volume of water three

times ihc weight of seeds for two hours. The soaked sceds wcre allowed to sprout at room

temperature for 24hr, 48hr, 72hr and 96hr. The seeds were watered three times a day according

to the method of Marero et al. (1988).

Fermen tation Procedure:

Whole seeds were weighed, washed and fermented for 24hr, 48hr, 72hr, and 96hr. The

fermentation process was by endogenous microflora in the grains at room tcrnpcraturc as

described by Onuoha and Obizoba, (2001).

Preparation of flour:

The fermented and germinated seeds were steamed for 30mins from the time of boiling

and dried in the oven at 100°C for about 8hr, after which, the dried seeds were cleaned to remove

the rootlets and hulls. The cleaned seeds were milled and sieved by passing through a Imin pore

size sieve to yield flour (Fig. 2 and 3). The flour was packed in thick polyethylene bags, sealed

and stored under reduced temperature (4OC) until needed for formulation and analysis.

Millet

1 Cleaning

1

Millet Flour

Dry milling Drying

/

Millet \

Flour Sieving

1 Sprouted Millet

Flour Figure 2: Flow diagram for millet flour production

Pigeon Pea I *

Cleaning

~ r y milling / S l i n g

Fermentation y hn\ Steepin soaking (24,4 ,72,96hr)

Untreated Pigeon pea Flour Steaming k Sprouting 1

(24, 48, 2, 96hr)

Drying i

Steaming (1 OO°C, 8hrs)

1 Dry milling

( 3 0 r s ) Drying

. Sievmg 1 1 (1 OO°C 8hrs)

Devegetation

1 1 / Fermented Dry milling Pigeon pea

Flour S'eving I, II \

Sprouted Pigeon pea

Flour

Figure 3: Flow diagram for Pigeon pea flour production

3.2 Preparation of Blends

The cereal: legume combination was formulated from the pre-treated samples as shown

in Table 11 and Table 12. The nutritional quality of the different flour blends (Table 12) was

evaluated by rat feeding experiments, while other physico-chemical characteristics of flours were

also analyzed.

3.3 Analytical Methods

3.3.1 Proximate Analysis

3.3.1.1 Moisture Content (A.O.A.C., 1990).

The air oven method was used in the estimation of the moisture contents of the flours. A

5.0g weight of sample in pre-weighed moisture dishes was dried to constant weight at 100°C

with intermittent cooling to room temperature in a desiccator and weighing.

Calculation:

% Moisture = Initial weight -Final weight X 100

lnitial weight

3.3.1.2 Fat Content (AOAC, 1990)

The fat content of sample was determined according to AOAC (1990), method. A 5g

weight of sample was weighed into a thimble lined with a circle of filter paper. Thimble and

contents were placed into a 50ml beaker and dried in an oven for 6hr at 100 - 102OC. Thimble

and contents were transferred to extraction apparatus. Beaker was rinsed several times with

ethylether and the rinsings added to the thimble in the extraction apparatus. The sample

contained in the thimble was extracted with ethylether in a soxhlet extraction apparatus for 6 - 8hrs at a condensation rate of at least 3 -6 drops per second.

At the completion of the extraction, the fat extract from the extraction flask was

transfered into pre-weighed evaporating dish with several rinsings of ethylether. The evaporating

dish was placed in a hme hood and with the fan on to evaporate off'the ethylether until no odour

of it was detectable.

Table 11: Proportions of different flour treatment in the composite blends.

Samvle Code Ratio Percentage

UPF+ UMF 70.30 70% UPF; 30% UMF

UPF+ FSMF 70:30 70% UPF; 30% FSMF

UPF + SSMF 70:30 70% UPF; 30% SSMF

FSPF + UMF 70:30 70% FSPF; 30% UMF

FSPF + FSMF 70:30 70% FSPF; 30% FSMF

FSPF + SSMF 70:30

SSPF + UMF 70:30

SSPF + FSMF 70:30

SSPF + SSMF 70:30

70% FSPF; 30% SSMF

70% SSPF; 30% UMF

70% SSPF; 30% FSMF

70% SSPF; 30% SSMF

Control 1OO:O 100% Control

Comrn. Diet 1OO:O 100% Comm. Diet.

Key:

UPF - Untreated Pigeon pea flour

FSPF - Fermented Steamed Pigeon pea Flour

SSPF - Sprouted Steamed Pigeon pea Flour

UMF - Untreated Millet flour

FSMF - Fermented Steamed Millet Flour

SSMF - Sprouted Steamed Millet Flour

Control - Casein -casein

Comm. Diet - Commercial Diet. 1'

The dish and contents were dried in an oven for 30minutes at 100°C. Dish plus contents \ \

were removed from oven, cooled in a desiccator and weighed. The ether extract was calculated \\

thus:

Calculation:

Crude fat (ether extract)% = (W2 - WL) X 100

S

Where:

W1 - Weight of empty evaporating dish

W2 - Weight of evaporating dish plus contents after drying

S - Sample weight (g).

3.3.1.3 Ash Content (AOAC, 1990)

A 5.0g weight of the sample was weighed into a weighed porcelain dish. The dish and

contents were transferred to a muffle hrnace and ignited at 500 to 600°C for about 8hr to a

greyish white ash. The dish and contents were removed from muffle hrnace, allowed to cool in a

desiccator and weighed. Result was calculated and expressed as % Ash.

Calculation:

Ash % = (B -C) X 100

A

Where

A - Sample weigh (g)

B - Weight (g) of dish and contents after drying

C - Weight (g) of empty dish

3.3.1.4 Crude Protein Determjnation (Nitrogen X 6.25) (AOAC, 1990)

A 2g weight of each sample was weighed into a digestion tube. A 15g weight of catalyst

(CuSo4: Na2So4 at 1: 10) and 35ml of concentrated sulphuric acid were added. The digestion of

each sample was carried out and digestion stopped when solution was clear.

Tubes and contents were allowed to cool to room temperature. The clear digest of each

sample was transferred-into a lOOml volumetric flask with several rinsings of distilled water and

made up to the mark when the solution cooled. A 5ml volume of the diluted digest was pipetted

and transferred to the distillation apparatus and distilled for 5 minutes after adding 5ml of O.IN

&So4 solution with 6 -8 drops of mixed methylredlmethylene blue indicator (1:l) and 5ml of

40% NaOH.

The distillate was titrated against 0.02N NaOH solution to a permanent green end-point.

A black titration was carried out by titrating 5ml of O.lN H2S04 with 0.02N NaOH.

Calculation:

Total Nitrogen % = (B - S) x 1.4007 x N x 20

Sample weight (g)

Where:

B = vol (ml) of NaOH solution used for black

S = vol (ml) of NaOH solution used for sample

N = Normality of NaOH

1.4007 = m.eq wt. of nitrogen (includes factors of 100 for %)

20 = Dilution factor (5ml of digest used out of 100ml).

Crude Protein % = Total Nitrogen % x 6.25

3.3.1.5 Crude Fibre Determination (AOAC 1990)

Crude fibre was determined according to AOAC, (1990) method. A 3.0g weight of the

sample was weighed into a 1L conical flask, defatted by Soxhlet extraction and hydrolysed with

200ml of boiling 0.255N H2S04 for 30mins on a hot plate. The flask was removed from heat,

allowed to settle and decanted through a Buchner fimnel, applying gentle suction. The residue on

filter paper was washed into the flask with hot 0.313N NaOH solution and the filter paper

washed with 1% HCL to make it acidic. The remaining particles were transferred from conical

flask to the paper with water and the paper was washed with water until acid -free. The paper

was later washed with alcohol and diethyl ether until all water is removed. The residue was air

dried and transferred to previously incinerated and weighed crucible with the aid of a spatula.

Then, later dried in the oven, cooled and weighed. The loss in weight represented the crude fibre /

content.

% Fibre = Loss in weight fiom incineration X 100 \ \

Weight of sample before defatting

3.3.1.6 Nitrogen Free Extract Determination

Nitrogen free extract content was determined by the difference in the sum of the

percentage moisture, fat, crude protein, ash and crude fibre from 100.

Calculation:

NFE=l00 - ( a + b + c + d - k e )

Where: a = Moisture

b = Fat content

c = Protein content

d = Ash content

e = Crude fibre

3.3.1.7 Caloric Value Determination The caloric value was determined using the Atwater quantification of the caloric (energy)

content of food as 4.0, 4.0 and 8.9 kcallg from carbohydrate, proteins and fat respectively (lkcal

= 4.186KJ) (Widdowson, 1987).

3.3.1.8 Determination of True Protein

The true protein content of the sample was determined according to the method of

McDonald et al. (1973). A 50g weight of sample was subjected to heat coagulation to separate

non-protein nitrogenous compounds from true protein. The solution was then filtered and the

residue was subjected to Kjeldahl determination of protein (AOAC, 1990). A 2g weight of

sample was weighed into digestion tube and digested with 15g of catalyst (CuS04: Na2S04 at

1:lO) and 35ml of concentrated sulphuric acid, until sample solution became clear. Tubes and

contents were allowed to cool to room temperature. The clear digest was transferred into a 1001nl /I

volumetric flask with several rinsings of distilled water and made up to the mark when the

solution cooled. A 5ml volume of the diluted digest was pipetted and transferred to the i ., distillation apparatus and distilled for 5 minutes after adding 51111 of 0. IN H2SO4 solution with 6 \

- 8 drops of mixed methylredlmethylene blue indicator (1 :I) and 5ml of 40% NaOH.

The distillate was titrated against 0,.02N NaOH solution to a permanent green end-point.

A blank titration was carried out by titrating 5ml of 0. IN H2S04 with 0.02N NaOfI. The protein

content was calculated thus.

Calculation:

Total Nitrogen % = (B-S) X 1.4007 X N X 20

Sample Weight (g)

Where:

B = Vol (ml) of NaOH solution used for blank

S = Vol (ml) of NaOH solution used for sample

N = Normality of NaOH

1.4007 = m.eq wt. of nitrogen (includes factors of 100 for %).

20 = Dilution factor (5ml of digest used out of 100ml).

True protein % = Total Nitrogen % X 6.25.

3.3.1.9 Mineral Content Determination

The mineral content was determined using the wet digestion method as described by

Adyeye and Ajewole (1992). A 2g weight of sample was weighed into the beaker, 25ml HN03

and 5ml perchloric acid were added, covered with wash glass and allowed to stand. The sample

solution was heated till it became clear. A lOml volume of 6MHCL was added to the sample

solution and the mixture boiled for 5mins. The content was quantitatively transferred into 50ml

volumetric flask and made up to the mark. The content was then filtered.

The filtrate was used to analyze for Ca, Fe and Zn. The concentration of each mineral

was determined by its capacity to absorb light of a defined warelength while in the vapour state

with atomic absorption spectrophotometer. Dilutions were made where possible. Readings of

varelength were plotted against concentration.

Calculation:

p of the curve X dilution (if any) X original vol. of digest X 100%

Weight of the sample X lo6

3.3.2 Determination of Antinutritional Factors

3.3.2.1 Determination of Hydrogen Cyanide Level.

Hydrogen cyanide level was determined using the quantitative method described by

lkediobi et al. (1980).

Sample extract was obtained by homogenizing 0.5g of sample with 32ml of distilled

water for 3min. The extract was centrifuged at 500 x g for 3minutes and the resulting supernatant .

used for analysis.

Linamarase Preparation: Thin brown outer skin of freshly harvested cassava tubers was

carehlly removed to reveal the underlying cortex. The cortex was removed and sliced into small

pieces. A lOOg of the sliced cortex was homogenized in 300ml of pre-chilled container and

filtered by suction through a lcm layer of kieselguhr. Filtrate was used to extract a second lOOg

batch of sliced cassava cortex as described above. The filtrate fiom the second extraction was

used to extract a third lOOg batch. This final extract was kept overnight at 4°C. About 2.3

volumes of cold acetone were added to the extract and solution swirled for 2 minutes several

times within a 2hr period. A pale green precipitate was recovered upon decantation. This

precipitate was extracted thrice with lOml portions of cold 0.1M acetate buffer of pH 5.5. The

extract was centrifuged at 500 x g for 5min at room temperature and the resulting supernatant

taken as the crude linamarase extract.

Quantitative Determination of Cyanide: A 0.5ml volume of sample extract was incubated

with I .Om1 of linamarase preparation for lominutes at room temperature in a tall-stoppered test

tube. The volume of the incubation mixture was made up to 2nd with 0.2M Sodiumphosphatc

buffer of pH 6.8. At the end of the incubation period, 5ml of alkaline picrate was added and

resulting solution was incubated in a water bath at 95OC for 5minutes. Following cooling to room

temperature, absorbance of the deip orange colour formed was read at 400nm in a

spectrophotometer. The cyanide concentration was extrapolated fiom a standard curve

previously prepared.

3.3.2.2. Determination of Phytate Level

The phytate level was determined by the method described by Thompson and Erdman

(1982). A 0.5g sample was placed into a 500ml flat-bottomed flask, into which 100.0ml of 2.4%

HCL was added. The flask was stoppered and shaken for lhr at room temperature on a

mechanical shaker. The extract was decanted and filtered. A 5ml volume of the filtrate was

pipetted into a 50ml beaker, which was diluted to 25ml with distilled water. A IOml volume of

the diluted sample was pipetted and 15ml of O.1M sodium chloride was added to it. The solution *

was passed through a 200 - 400 mesh to remove inqganic phosphorus and 15ml of O.7M sodium

chloride was added to separate phytate. Absorbance was read at 500nm by using water to zero

the spectrophotometer. A calibration curve was constructed and was used to calculate the

concentration of the phytate.

Calculation:

X x dilutions (if anv) x original vol. of digest x 100%

Weight of the sample x lo6

Where:

X = Value of the curve or optical density x reading from spec.

3.3.2.3. Determination of Tannin Levels

Tannin level was determined by the method described by Price and Butler (1977). A

60mg weight of ground grain was shaken constantly for 60s with 3ml of methanol in a test tube

and poured into a Buchner hnnel with the suction already turned on. The tube was quickly

rinsed with an additional 3ml of methanol and content was poured at once into the filnnel.

Filtrate was mixed with 50ml of water and analyzed within an hour. A 3nd volume of 0.1M

FeCL3 in 0. IN HCL was added to the extract and 3ml of 0.008MK#c (CN)G was added. l'hc

optical density was read aRer lominutes in lcm glass cells at 720nm in a spectrophotometer that

was zeroed with water. Results were extrapolated from tannin standard curve.

3.3.3. Functional Properties ~etermination

3.3.3.1. Particle Size Distribution

The method described by Ihekoronye and Oladunjoye (1988), was used in the

determination of particle size. A nest of six sieves (US 20, 30, 40, 50, 100 and 230 mesh screens,

corresponding to sieve openings of 850, 600, 425, 300, 150 and 63pm) was arranged in \ \

descending order. The upper sieve was provided with a cover and the bottom sieve with a

receiver. A 50g sample of the product was put in the top sieve, covcred with the lid and the nest

of sieves placed in a suitable mechanical sieve shaker. The material was sieved continuously for

5minutes and stopped. The nest was removed and the residue on each sieve transferred to a

tarred weighed dish using a brush. Each dish was weighed and the percentage of product retained

on each sieve was calculated as:

Mass of material retained on specific sieve x 100 Total mass of sample

3.3.3.2. Viscosity

The viscosity of porridges from sample flour was determined according to the method

described by Sathe and Salunkhe (1981), using the Gallenkamp Universal Torsion Viscometer.

The viscometer is composed of a vertical torsion wire, a flywheel mounted above a graduated

scale and a cylinder suspended below the scale. Each sample was prepared at concentration of

2,4,6 and 10% ("/,). A 5ml of sample solution was poured into the viscometer cup. The flywheel

was rotated through 360" and then released. The damping effect of sample on the overswing of

the cylinder gave a measure of its viscosity. Porridges were kept in thermoflask to maintain the

serving temperature (40°C) during the determination. Viscosity was calculated relative to

distilled water as follows:

Where:

V1 = Viscosity of water at room temperature

VZ = Viscosity of sample to be determined.

dl = Density of water at room temperature.

d2 = Density of sample at room temperature.

tl = Flow rate of water at room tkmperature.

t2 = Flow rate of sample at room temperature.

3.3.3.3. Water Absorption Index

The method described by Onwulata el a/., (1 998), was used in the determination of water \

absorption index. A 1 .Og weight of sample was placed in a centrihge tube and lOml distilled

water was added. After standing for 15mins with intermittent shaking (5mins), sample solution

was centrihged for 15minutes at 1000 x g. Supernatant was decanted and the weight gain in gel

noted. Water Absorption Index was calculated as the weight gain of the gel dry weight.

Calculation:

WAI (Water Absorption Index) = W2- WI

Mass of material retained on specific sieve x 100 Total mass of sample

3.3.3.2. Viscosity

The viscosity of porridges from sample flour was determined according to the method

described by Sathe and Salunkhe (1981), using the Gallenkamp Universal Torsion Viscometer.

The viscometer is composed of a vertical torsion wire, a flywheel mounted above a graduated

scale and a cylinder suspended below the scale. Each sample was prepared at concentration of

2,4,6 and 10% ("I,). A 5ml of sample solution was poured into the viscometer cup. The flywheel

was rotated through 360" and then released. The damping effect of sample on the overswing of

the cylinder gave a measure of its viscosity. Porridges were kept in thermoflask to maintain the

serving temperature (40°C) during the determination. Viscosity was calculated relative to

distilled water as follows:

Where:

V1 = Viscosity of water at room temperature.

VZ = Viscosity of sample to be determined.

dl = Density of water at room temperature.

d2 = Density of sample at room temperature.

tl = Flow rate of water at room timperature.

tz = Flow rate of sample at room temperature.

3.3.3.3. Water Absorption Index

The method described by Onwulata et a/., (1998), was used in the determination of water \

\ absorption index. A 1.0g weight of sample was placed in a centrifbge tube and lOml distilled

water was added. After standing for 15mins with intermittent shaking (5mins), sample solution

was centrifbged for 15rninutes at 1000 x g. Supernatant was decanted and the weight gain in gel

noted. Water Absorption Index was calculated as the weight gain of the gel dry weight.

Calculation:

WAI (Water Absorption Index) = W2- W1

Where:

W1 = Original weight of dry sample

W2 = Weight of sample after absorbing water.

3.3.3.4. Water Solubility Index

Water solubility index was determined by modifLing the method reported by Onwulata et

al., (1998). A 2g weight of sample was weighed into a weighed porcelain containing lOml of

water. The dish and content were heated in a water bath at 100°C for 30minutes. The slurry was

allowed to stand and cool to room temperature. The supernatant was decanted, evaporated to

dryness and weighed. Water solubility index was calculated as:

Where:

W1 = Weight of original sample

W2 = Weight of dry matter

3.3.3.5. Least Gelation Concentration

The least gelation concentration of flour samples was determined according to the

method described by Ihekoronye (1986). Appropriate sample suspensions of 2, 4, 6, 8, 10, 12,

14, 16, 18 and 20% (wlv) were prephred in 5ml distilled water. Test tubes containing these

suspensions were heated for lhr in a boiling water batb and then rapidly cooled under running

cold tap water. The test tubes were hrther cooled for 2hr at 4°C. Least gelation concentration

was determined as that concentration when the sample from the inverted test tube would not fall /' I

down or slip.

3.3.3.6. Bulk Density Determination

The bulk density was determined by the method described by Nwanekezi et al., (2001).

Each sample was slowly filled into 5ml calibrated cylinders. The bottom of the cylinder was

gently tapped on a laboratory bench until there was no further lessening of the sample after

filling to 5ml mark. Bulk density was estimated as mass per unit volume of the sample taken.

The mean of the triplicate measurements was taken as the estimate of bulk density.

Calculation:

Bulk density (glml) = Mass of sample

Volume occupied by sample

3.3.3.7. Reconstitution Time Determination

Reconstitution time determination was carried out using the method described by

Nwanekezi et al., (2001). A 2g weight of each sample was sprinkled on the surface of 50ml of

cold distilled water contained in 150ml cylinder. The time taken for each of the samples to

completely dissolve with stirring was recorded. The mean value of triplicate determinations was

taken.

3.3.4. Sensory Analysis

Laboratory preference test was done using a 30-man panel of judges, who ranked the

products using a seven point hedonic scale as described by Land and Shepherd, (1984). The

preference test was aimed at selecting the organoleptically preferred formulations.

3.3.5 Animal Bioassay

A 35 -day study that included a 28 -day growth and a 7 -day N balance period was

carried out. Forty-two (42) weanling dale rats weighing 45-556 were used for the study. The rats

were divided into 7 groups of six rats each on the basis of body weight. The animals werc

weighed prior to access to the test diets and at weekly intervals to determine the gain in body

weight. The rats were housed in individual metabolic cages and fed he diets and tap water for 35

days. Table 12 contains the composition of the experimental diets. The animals were fed their

respective diets for 28 days and their growth was determined. The same diets and groups of rats

were fed for an extra one week to study their N balance. Carmine red was fed on the morning of

day 28 and of day 35. The coloured feaces that appeared on day 29 were included in the pooled

feacal sample while those feaces that appeared on day 36 were excluded. Urine was collected

from morning of day 29 through to the morning of day 36 (7 days) and food consumption was

measured for the same 7 -day period for all groups of rats. HCL (Hydrochloric acid) (0.IN)

(Iml) was added and used as a preservative for individual urine collection (Okeke and Obizoba, , 1986).

Calculation:

Bulk density (g/ml) = Mass of sample

Volume occupied by sample

3.3.3.7. Reconstitution Time Determination

Reconstitution time determination was carried out using the method described by

Nwanekezi et al., (2001). A 2g weight of each sample was sprinkled on the surface of 50ml of

cold distilled water contained in 150ml cylinder. The time taken for each of the samples to

completely dissolve with stirring was recorded. The mean value of triplicate determinations was

taken.

3.3.4. Sensory Analysis

Laboratory preference test was done using a 30-man panel of judges, who ranked the

products using a seven point hedonic scale as described by Land and Shepherd, (1984). The

preference test was aimed at selecting the organoleptically preferred formulations.

3.3.5 Animal Bioassay

A 35 -day study that included a 28 -day growth and a 7 -day N balance period was

carried out. Forty-two (42) weanling hale rats weighing 45-55g were used for the study. The rats

were divided into 7 groups of six rats each on the basis of body weight. The animals were

weighed prior to access to the test diets and at weekly intervals to determine the gain in body

weight. The rats were housed in individual metabolic cages and fed he diets and tap water for 35

days. Table 12 contains the composition of the experimental diets. The animals were fed their

respective diets for 2.8 days and their growth was determined. The same diets and groups of rats

were fed for an extra one week to study their N balance. Carmine red was fed on the morning of

day 28 and of day 35. The coloured feaces that appeared on day 29 were included in the pooled

feacal sample while those feaces that appeared on day 36 were excluded. Urine was collected

from morning of day 29 through to the morning of day 36 (7 days) and food consumption was

measured for the same 7 -day period for all groups of rats. HCL (Hydrochloric acid) (0.1N)

(Iml) was added and used as a preservative for individual urine collection (Okeke and Obizoba, , 1986).

Table 12: Composition on drv wei~ht basis (d100g) of millet, pigeon pea and

casein diets fed to rats.

Diets I I1 I11 IV V VI VII Sources of Protein and ratios

Diet UPF:

UMF

Composition 70:30 U P F 800.35 U M F 823.54

F S P F - S S P F - F S M F - S S M F - Casein - Comrn. Diet - Oil 126

Mineral 88.2

Vitamin 25.2

Fibre 25.2

Corn Starch 3 15.755

Sucrose 315.755

FSPF:

FSMF

70:30 - -

75 1.97

-

799.09

-

-

- 126

88.2

25.2

25.2

352.17

352.17

Total 2520 2520 2520 2520 2520 2520 2520

Key:

UPF: - Untreated Pigeon pea flour

UMF: - Untreated Millet Flour

FSPF: - Fermented Steamed Pigeon pea I%x~r

SSPF: - Sprouted Steamed Pigeon pea flour

FSMF: - Fermented Steamed Millet Flour

SSMF: - Sprouted Steamed Millet Flour

Comm. Diet: - Commercial Diet

Casein: - Casein control.

FSPF:

SSMF

70:30 - -

75 1.97

- -

959.36

- -

126

88.2

25.2

25.2

272.035 . 272.035

SSPF:

FSMF

70:30 - - -

707.1 1

799.09

- -

-

126

88.2

25.2

25.2

374.6

374.6

SSPF:

SSMF

7 0 3 0 - -

-

707.1 1

-

959.36

-

- 126

88.2

25.2

25.2

294.465

294.465

Casein

1oo:o - - -

- - -

302.88

- 126

88.2

25.2

25.2

976.26

976.26

At the end of the feeding experiment, N balance, body weight, protein efficiency ratio

(PER), N retention, net protein utilization (NPU) and biological value (BV) were assayed using

the method described by Ihekoronye and Ngoddy, (1 985).

3.3.6 Statistical Analysis

Data were statistically analyzed using analysis of variance (ANOVA) and least

significant difference (L.S.D) to test the differences among the nutritional value of protein

mixtures. Similarly, protein mixtures were statistically analyzed to determine which protein

mixtures were significantly different in N intake, weight gain, N retention, biological value (BV)

and other parameters. The package used for the analysis is GenStat Release 4.23DE (PC\window

98).

CHAPTER FOUR

4.0 Results and Discussions

4.1 Nutrient Composition

4.1.1 Effect of Fermentation on Proximate Composition of Millet and

Pigeon Pea Flours:

Table 13 shows the proximate composition of the unfermented and fermented millet and

pigeon pea flours. The crude protein content of the unfermented millet was 9.18% while for the

various fermentation regimes, the protein contents were 7.45% (24hr), 8.19%(48hr),

9.46%(72hr) and 8.9%(96hr). Fermentation for 24hr and 48hr significantly (P<0.05) decreased

the crude protein level by 18.85% and 10.78% respectively. Furthemlore, fermentation for 96hr

showed a non -significant decrease in the crude protein level by 3.05% of that originally present

while 72hr fermentation showed a non-significant (P>0.05) increase in the crude protein level of

millet by 3.05% of that originally present. The increase in crude protein content of the 72hr

fermented millet suggests that 72hr is the optimum fermentation period for protein synthesis by

the sample fermenting microflora. The decrease in crude protein in the 96hr fermentation regime

is not in agreement with the observation of Obizoba and Egbuna (1992), who reported increase

in the protein content of bambara groundnut after 96hr fermentation. The protein value for the

unfermented pigeon pea was 22.04% while for the various fermentation regimes, the protein

contents were 18.40%(24hr), 17.5 1 %(48hr), 23.46%(72hr) and 18.55%(96hr). Fermentation for

24hr, 48hr and 96hr decreased the crude protein content of the pigeon pea by 16.52%, 20.55%

and 15.84% respectively, while the 72hr fermentation regime increased the protein content by

about 6.44%. The high protein content of 72hr-fermented pigeon pea could be due to the release

of free amino acids for protein synthesis (Egbekun, 1998). The decrease in protein content at

96hr fermentation could be attributed to the fact that microorganisms used the nutrient for their

metabolism and growth (Ariahu et. al., 1999). A similar increase in protein content of limabcans

after 48hr fermentation was reported by Onuoha and Obizoba (2001). This was followed by a

reduction in value as fermentation proceeded. The fat content of the fermented millet varied from

3.7% at 24hr, 2.1% at 48hr, 4.5% at 72hr to 3.53% at 96hr, while that of unfermented rnillet was

1.5%. Fermentation significantly (K0.05) increased the fat content of fermented millet ('Table

13). 9

Table 13: E

ffect of fermentation tim

e on proximate com

position of millet and pipeon pea flours (%

dry weight

basis)

Sa

m~

les Crude P

rotein C

rude fat C

rude fibre M

oisture content A

sh N

itropen Free E

xtract E

nerev

LSD

(0.05) 0.957

0.14 0.14

0.17 0.32

1.002 2.901

Mean values of triplicate determ

inations

UM

F -Unferm

ented Millet Flour

UPF -

Unferm

ented Pigeon pea Flour

FMF24 - 24hr Ferm

ented Millet Flour

FPF24 - 24hr Ferm

ented Pigeon pea Flour

FMF48 -

48hr Fermented M

illet Flour FPF41 -

48hr Fermented Pigeon pea Flour

FMF72 -

72hr Fermented M

illet Flour FPF72 -

72hr Fermented Pigeon pea Flour

FMF96 -

96hr Fermented m

illet Flour FPF96 -

96hr Fermented Pigeon pea Flour

Furthermore, the fat content of fermented pigeon pea also varied from 0.8% at 24hr, 0.5% at

48hr, 1.6% at 72hr to 1.3% at 96hr. The fat content of the unfermented pigeon pea was 2.7%.

Fermentation significantly (P<0.05) decreased the fat content of pigeon pea. The low level of fat

at 48hr fermentation (2.1%) for millet and for all fermentation periods for pigeon pea could be

due to the activities lipolytic enzyme which hydrolyzed the fat to glycerol and fatty acids. The

free fatty acids might have reacted with other products of hydrolysis to form esters resulting in a

decrease (Nnam, 2001). However, the increase in the fat content of millet with fermentation

could be attributed to increased activity of the lipolytic enzymes of the fermenting organisms in

releasing free fatty acids. This result compares with results reported by Obizoba and Atii (1994).

They reported an increase in fat content of millet with fermentation. The fermented samples

showed comparable (P>0.05) crude fibre content, which varied from 2.8% to 3.0% in millet and

1.8% to 2.1% in pigeon pea. The moisture level after 72hr fermentation of millet (6.6%) was

significantly (P<O.OS) lower than those of the other millet samples fermented for other periods.

Likewise, the moisture content (9.0%) of the 48hr fermented pigeon pea was also significantly

lower (P<O.OS) than those of other fermented pigeon pea sample. Moisture content of food

products are affected by different factors such as the type and amount of proteins and the type of

treatment given to the products. These factors may also have influenced the moisture content

observed in this study. There is a variation in the ash level of the fermented samples. The

carbohydrate level of fermented millet ranged from 71.20% to 75.49%. Millet fermented for 72hr

showed a significantly (P<O.OS) high carboliydrate value than the other millet samples fermented

for 24hr and 96hr. The carbohydrate level of fermented pigeon pea ranged from 61.36% to

69.92%. The 48hr fermented pigeon pea showed a significantly (P<0.05) higher carbohydrate

value compared to others. The reduction in carbohydrate content after 72hr fermentation in millet

and after 48hr fermentation in pigeon pea could be attributed to possible hydrolysis of complex

carbohydrate to simple sugars, which were used for metabolic processes (Nnam, 2001). Obizoba

and Atii (1991) reported a similar decrease in carbohydrate level of sorghum after 48hr

fermentation.

4.1.2 Effect of Sprouting on Proximate Composition of Millet and Pigeon

Pea Flours

The proximate composition of unsprouted and sprouted millet and pigeon pea is shown in

Table 14. The protein values for various sprouted millet samples were 7.88%(24hr),

7.45%(48hr), 7.59%(72hr) and 7.88%(96hr), which were significantly (P<0.05) lower than the

protein value for the unsprouted millet (9.18%). Sprouting for 24hr, 48hr, 7 2 1 ~ and 96hr

decreased the protein value by l4.16%, l8.85%, I 7.32% and 14.16% respectively. The protein

content of the unsprouted pigeon pea was 22.04% and for all the sprouted pigeon pea, the protein

values were 24.85%(24hr), 18.1 O%(48hr), 2 I .16%(72hr) and 20.14%(96hr). Pigeon pea sprouted

for 24hr had a significantly (P<0.05) higher protein content than pigeon pea sprouted for other

periods and the unsprouted pigeon pea (Fig 5). This high protein content at 24hr sprouting could

be attributed to the release of free amino acid for synthesis of protein as a result of the

breakdown of tannin -protein complexes (Chavan et al., 198 1 ). The reduction in protein value at

48hr sprouting could be attributed to the loss of soluble protein and other nitrogenous

constituents in the soak water as observed by Obizoba and Atii (1991). It could also be attributcd

to the hydrolysis of protein to amino acids, which were then used by the sprouting shoots or

embryos for growth. In a previous study, Onimawo and Asugo (2004) reported a decreasc in

crude protein content of germinated pigeon pea. Similarly, Obizoba and Egbuna (1 992) reported

a decrease in protein content of gepninated bambara groundnut. Akubor and Obiegbuna (1999) '

reported that germination decreased the crude protein content of millet flour. 'I'he fat content

(Table 14) of sprouted pigeon pea was significantly (P4.05) reduced as the sprouting days

increased. The observed decrease could most likely be due to the activities of lipolytic enzymes,

which hydrolyzed, fat to glycerol and fatty acids, which form esters with other products of i \ hydrolysis bringing about a decrease. Previous study by Onimawo and Asugo (2004), also \

reported a decrease in the fat content of germinated pigeon pea. The fat content of millet

sprouted for 48hr (0.9%) was significantly (P<0.05) lower than the fat content of other san~ples

3.1% (24hr), 2.3% (72hr) and 2.1% (96hr). The increase in fat content after 48hr sprouting was

attributed to the synthesis of new lipids by microflora during metabolic activities (Nnam, 2001).

This observed increase in fat content compares well with the result of Obizoba and Atii (1991),

who reported an increase in fat content of sorghum due to germination. The crude fibre of 1

content sprouted millet varied from 1.8% to 3.2% while that of sprouted pigeon pea varied fioin

Table 14: E

ffect of sprou

t in^

time on proxim

ate composition of m

illet and pipeon pea flours (% dry w

ei~h

t

basis).

Sa

m~

les C

rude Protein

Crude fat

Crude fibre

Moisture content

Ash

Nitropen F

ree Extract

Enerw

UM

F 9.18

1.5 3.0

6.8 3.7

75.82 353.2

SMF24

7.88 3.1

3.2 8.2

3.7 73.88

353.3

SMF48

7.45 0.9

3.1 10.1

2.8 75.62

340.4

sW72

7.59 2.3

1.8 10.2

3.3 74.84

350.7

SMF96

7.88 2.7

2.6 6.8

3.9 76.62

357.2

UPF

22.04 2.7

2.2 9.7

1.3 62.06

360.4

SPF24 24.85

1.8 2.3

10.5 2.6

57.85 347.1

SPF48 ,

18.10 1.2

2.1 11.1

1.1 66.37

349.0

SP

Fn

21.16 1.1

1.7 9.6

2.1 64.24

351.8

SPF96 20.14

0.2 2.2

12.5 1.2

63.79 337.2

LSD

(0.05) 1.204

0.1 1 0.17

0.17 0.32

1.193 1.86

Mean values of triplicate determ

inations

UM

F -Unferm

ented Millet Flour

UPF -

Unsprouted Pigeon pea Flour

SMF24 -

24hr Sprouted Millet Flour

SPF24 - 24hr Sprouted Pigeon pea Flour

SMF48 - 48hr Sprouted M

illet Flour SPF48 - 48hr Sprouted Pigeon pea Flour

SMF72 -

72hr Sprouted Millet Flour

SPF72 - 72hr Sprouted Pigeon pea Flour

ShfFg6 - 96hr Sprouted m

illet Flour - 96hr Sprouted Pigeon pea Flour

1.7% to 2.3%. The moisture content of the 96hr-sprouted millet (6.8%) was lower than the

24hr(8.2%), 48hr(lO. 1%) and 72hr(10.2%). Likewise, the moisture content of pigeon pea '

sprouted for 72hr(9.6%) was also lower than those sprouted for 24hr(10.5%), 48hr(11 .I%) and

96hr(12.5%). Different factors affect moisture content of food products. The variation in

moisture content might be attributed to treatments, which caused changes in other nutrient

contents. The ash content varied amongst the samples. The carbohydrate value of pigeon pea was

increased by sprouting except for the 24hr-sprouted pigeon pea, which was significantly

(Pc0.05) low (57.85%), while the sprouted millet samples had comparable (P>0.05)

carbohydrate content with the unsprouted millet. The change in carbohydrate content was

attributed to the metabolic activities of the hydrolytic enzymes within the seeds during sprouting

(Nnam 2001).

4.1.3 Effect of Fermentation on Antinutritional Content of Pigeon Pea

and Millet

Table 15 shows the antinutrient content of unfermented and fermented millet and pigeon

pea. The tannin level of the unfermented millet was 2.80mdg and for the fermented millet the

values were 2.57mg/g (24hr), 2.56mglg (48hr), 2.49mgl (72hr) and 3.20mglg (96hr).

Fermentation had varied effects on the tannin level of millet when compared to the unfermented

millet. Furthermore, fermentation had no significant effect on the tannin level of fermented

millet. The tannin level of the unfermented pigeon pea was 2.7mglg while for the fermented

pigeon pea the values were 2.45mg/g (24hr), 2.60mglg (48hr), 2.26mglg (72hr) and 2.47mglg

(96hr). The tannin level of the fermented pigeon pea was lower than that in the unfermented

pigeon pea. However, fermentation of pigeon pea for 72hr significantly (Pc0.05) decreased the

tannin level. There was no significant (P>O.OS) difference in the tannin level of pigeon pea

fermented at other periods. The decrease observed in the fermented pigeon pea was as a result of

the treatment in which enzymes were produced which brokedown complexes to release fiee

tannins. The fiee tannins released were leached out resulting in a decrease (Obizoba and Atii,

1991). The phytate level of unfermented millet was 0.01 17mglg and the fermented millet varied

between 0.0044mglg (24hr), 0.0027mdg (48hr), 0.0028mglg (72hr) and 0.001 8mdg (96hr),

D b l e 15: Effect of fermentation on antinutritional content of pipeon pea and millet.

Sarnnle Tannin (rndp;) Phytate (rnejg) Cyanide (rndg)

UMF 2.80 0.01 17 4.9

FMF24 2.57 0.0044 4.9

FMF4 8 2.56 0.0027 5.1

FMF72 2.49 0.0028 4.9

FMF96 .3.20 0.00 1 8 5.1

UPF 2.70 0.1 177 4.7

FPF24 2.45 0.0360 4.9

FPF48 2.60 0.0073 5.5

FPF72 2.26 0.0240 4.5

FPF96 2.47 0.0323 4.5

LSD (0.05) 0.4 15 0.00536 0.30

Mean values of triplicate determinations

UMF -Unsprouted Millet Flour UPF -Unsprouted Pigeon pea Flour

SMF24 -24hr Sprouted Millet flour SPF24 -24hr Sprouted Pigeon pea Flour

SMF48-4hr Sprouted Millet Flour SPF48 -48hr Sprouted Pigeon pea Flour

SMF72 -72hr Sprouted Millet Flour SPF72 -72hr Sprouted Pigeon pea Flour

SMFg6 -96hr Sprouted Millet Flour SPF96 -96hr Sprouted Pigeon pea Flour

However, fermentation significantly (P<0.05) reduced phytic acid content of millet by a

factor of 62.39%(24hr), 76.92%(48hr), 76.07%(72hr) and 84.62%(96hr) when compared to the

unfermented millet. The phytate level of the fermented pigeon pea ranged from 0.0073mglg to 1 0.036mglg which was significantly (W0.05) lower than the value (0.1 177mglg) in the

unfermented pigeon pea (Fig 6). The reduction in the phytate content of the fermented millet and \

pigeon pea could be attributed to the activity of the microflora, which brokedown organic

complexes in the samples releasing phytate which was leached out into the surrounding

fermentation medium (Onuoha and Obizoba, 2001). Ariahu et al. (1999) reported a similar

decrease in phytate level of fermented soybean and breadfruit. Fermentation had varied effect on

the cyanide level of the fermented millet and pigeon pea. The cyanide level of unfermented

millet was 4.9mglg and the cyanide levels of the fermented millet ranged fiom 4.9mglg to

S.lmg/g (Table 15). The cyanide level of the fermented millet was not significantly (P>0.05)

different from the cyanide level in the unfermented millet. The cyanide level of the unfermented

pigeon pea was 4.7mg/g, while the cyanide levels of the fermented pigeon pea varied from

4.9mg/g (24hr), 5.5mg/g (48hr), 4.5mg/g (72hr) and 4.5ingJg (96hr). There was a significant

(P<0.05) increase in the cyanide level of the 48hr fermented pigeon pea, while fermentation at

other periods showed no significant (P>0.05) difference from the unfermented pigeon pea. The

increase in the cyanide levels could be attributed to the increased enzyme activity which led to

increased hydrolysis of more cyanogenic glycosides to HCN and reduced elimination of the

HCN (cyanide) from the system. It could also be that fermentation of raw seeds was not effective

in removing HCN which is distributed through out the kernel. Obizoba and Atii (1994), made

similar observation in the investigation they carried out on millet. The reduced level of cyanide

in pigeon pea could be attributed to the fact that fermentation increased the hydrolysis of

cyanogenic glucosides to cyanide by the activity of the enzyme present. The cyanide produced

was finally leached out (Obizoba and Atii, 1991).

4.1.4 Effect of sprouting on antinutrient content of millet and pigeon pea

Table 16 presents the antinutrient content of sprouted and unsprouted millet and pigeon

pea. 'The level of tannin in sprouted millet varied from 2.95mg/g (24hr), 4.05mg/g (48hr),

5.73mglg (72hr) to 5.12mglg (96hr), while the tannin level in the unfermented millet was t

2.80mglg. Sprouting significantly (P<0.05) increased the tannin level of millet sprouted for

different periods except for the millet sprouted for 24hr which showed no significant (P>0.05)

difference from the unsprouted millet. The tannin level for unsprouted pigeon pea was 2.7mglg

and the tannin levels for the sprouted pigeon pea were 2.38mg/g (24hr), 3.07mgJg (48hr),

2.7mg/g (72hr) and 2.82mg/g (96hr). The increase in tannin level was attributed to reduced

elimination of the released tannin, which then formed new complexes with protein, therefore

resulting in an increase (Chavan et al., 1981). The decrease in tannin level of the 24hr sprouted

pigeon pea could be attributed to the hydrolytic activity of enzymes inherent in the sprouting

seeds. The enzymes hydrolyzed the tannin -protein and tannin -enzyme complexes to remove

tannins. The free tannins was leached out (Farhangi and Valadon, 1981). It has also been

Table 16: Effect of sprouting on antinutritional content of millet and ~ i ~ e o n

Samples Tannin (m&) Phylate (mgjg) Cyanide (m&)

UPF 2.70 0.1 177 4.7

LSD (0.05) 0.407 0.00396 0.49

Mean values of triplicate determinations

UMF -Unsprouted Millet Flour UPF -Unsprouted Pigeon pea Flour

SMF24 -24hr Sprouted Millet flour SPF24 -24hr Sprouted Pigeon pea Flour

SMF48 -4hr Sprouted Millet Flour SPF48 -48hr Sprouted Pigeon pea Flour

SMF72 -72hr Sprouted Millet Flour SPF72 -72hr Sprouted Pigeon pea Flour

SMF96 -96hr Sprouted Millet Flour SPF96 -96hr Sprouted Pigeon pea Flour

observed that malting effectively lowers tannin value (Osuntogun et al., 1989), (Obizoba and

Egbuna, 1992). The phytate level of the sprouted millet ranged fiom 0.001Gmg/g to 0.0028mglg / /

and the phytate content of the unsprouted millet was 0.01 17mglg. The phytate content of the

sprouted pigeon pea ranged fiom O.Olmg/g to 0.063mglg while the phytate content of the \ \ \

unsprouted pigeon pea was 0.11 77mglg. Sprouting caused a significant (W0.05) decrease in the

phytate content of millet and pigeon pea. The decrease was attributed to the breakdown of

phytate complexes by phytase, releasing phytate which was then leached out, resulting in a

decrease. Singh (1991) also noted that sprouting can reduce or eliminate appreciable amounts of

phytic acid in legumes and hence improve mineral bioavailability. Similarly, decrease in phytate

level have been reported in germinated Faba beans (Youssef et al., 1987) and germinated soybean *r

and African breadfiuit (Ariahu et a!., 1999). Sprouted pigeon pea and millet had higher cyanide

level than the unsprouted pigeon pea and millet. The cyanide level in the sprouted millet ranged '

fiom 5.3mg/g to 7.3mg/g while in the unsprouted millet, the level was 4.9mdg. Sprouting

significantly (P<0.05) increased the cyanide level of millet sprouted for different periods except

for 24hr sprouted millet which showed no significant (P>0.05) increase when compared to the

unsprouted millet. The cyanide level of the sprouted pigeon pea ranged fiom 4.7mg/g to 5.3mg/g

while in the unsprouted pigeon pea, the level was 4.7mg/g. Sprouting, significantly (P<0.05)

increased the cyanide level of 96hr sprouted pigeon pea while there was no significant (P>0.05)

increase in cyanide content of samples from the other sprouting periods. The increases observed

in the cyanide content of sprouted samples could be due to the increased activity of the enzyme

inherent in the developing embryo. Sprouting activates the enzymes in these seeds to hydrolyze

cyanogens to hydrogen cyanide (HCN) (Obizoba and Atii, 1994).

Germination and fermentation could be beneficially used to improve the nutritional

quality of cereals and legumes by reducing the antinutritional level of the grains (Egbekun,

1998).

Based on the antinutrient content and the proximate composition of the sprouted and

fermented millet and pigeon pea, samples SMF24, FMF72, SPF24 and FPF72 were selected for

hrther studies and for product formulation.

4.1.5 Effect of Steaming on proximate Composition of Fermented and

Sprouted Millet and Pigeon pea

Table 17 shows the proximate composition of fermented steamed and sprouted steamed

millet and pigeon pea flours. The protein content of sprouted steamed and fermented steamed

millet was 8.57% and 7.85% respectively. Steaming significantly (P<0.05) reduced the protein

content of fermented millet by 14.49% but the 6.65% reduction in the protein content of the

sprouted millet was not significant statistically. Sprouted steamed pigeon pea and fermented

steamed pigeon pea had protein level of 16.84% and 18.63% respectively. Steaming significantly

(P<0.05) reduced the protein content of sprouted and fermented pigeon pea by 23.59% and

15.47% respectively. There was a reduction in the protein contents of both sprouted steamed,

fermented steamed pigeon pea and millet when compared to the untreated pigeon pea (22.04%) v

and untreated millet (9.18%). This reduction might be attributed to protein denaturation due to

steaming and also to the formation of tannin - protein complexes, which resulted in reduced .

availability of protein (Chavan et al., 1981). The lipid contents of the sprouted steamed and

fermented steamed millet (7.03% and 4.8% respectively) were significantly (P<0.05) higher than

the untreated millet (1.47%). Sprouted steamed and fermented steamed pigeon pea had

significantly (WO.05) higher lipid contents (5.90% and 5.53% respectively) than the untreated

pigeon pea (2.67%). This increase in the lipid level could be attributed to the increased activity

of the lipolytic enzymes, which increased the release of more free fatty acids that may have

influenced the flavour of the product (Obizoba and Atii, 1994). Steaming significantly (P<0.05)

reduced the crude fibre content of the sprouted and fermented millet. The crude fibre content of

fermented and sprouted pigeon pea was comparable (P>0.05) to the crude fibre content of the

untreated pigeon pea. Steaming significantly (PC0.05) increased the moisture content of sprouted

millet but had no significant effect on the moisture content of the fermented and sprouted pigeon

pea. Obizoba and Atii, (1994), similarly observed an increase in moisture content of cooked

sprouted millet. There was a variation in the ash content of the sprouted steamed and fermented

steamed pigeon pea. Steaming significantly (Pc0.05) reduced the ash content of sprouted and

fermented millet. The carbohydrate content of sprouted millet was significantly (Pc0.05) reduced

by steaming while fermented millet had comparable (P>0.05) carbohydrate content with thc

untreated millet. Steaming significantly (P<0.05) increased the carbohydrate content of sprouted

pigeon pea but significantly (P<0.05) reduced the carbohydrate content of fermented pigeon pea.

The decrease in carbohydrate content could be attributed to increased activity of amyloytic

enzymes inherent in the products, which hydrolyzed the carbohydrate to simpler and much more

utilizable sources of energy (Obizoba and Atii, 1994).

4.1.6 Effect of steaming on the mineral content of fermented and

sprouted pigeon pea and millet

The mineral content of fermented steamed and sprouted steamed pigeon pea and millet is

shown in Table 18. The calcium content of fermented steamed and sprouted steamed millet was

5.85rneJg and 0.57mglg respectively. Steaming significantly (Pc0.05) increased the calcium

content of fermented millet, while significantly (P<0.05) reducing the calcium content of w

sprouted millet. A similar decrease was observed iq sorghum sprouted for 48hr (Nnam, 2001).

Table 18. Effect of steaming on the mineral content of fermented and sprouted

pigeon pea and millet

Samples Calcium Iron Zinc

UMF

SSMF

FSMF 5.85 6.44 47.00

UPF 10.30 4.90 28.20

SSPF 5.00 6.68 44.00

FSPF 4.70 7.05 44.00

LSD (0.05) 0.395 . 1.274 3.368

Mean values of triplicate determinations

UMF - Untreated Millet Flour

SSMF - Sprouted Steamed millet flour

FSMF - Fermented Steamed Millet Flour

UPF - Untreated Pigeon pea Flour

SSPF - Sprouted Steamed Pigeon pea flour

FSPF - Fermented Steamed Pigeon pea flour

The calcium content of sprouted and fermented pigeon pea was 5.00mglg and 4.70mglg

respectively. Unlike in millet, steaming significantly (Pc0.05) decreased the calcium content of

both fermented and sprouted pigeon pea by 54.37% and 51.46% respectively, Onuoha and

Obizoba (2001), observed similar decrease in calcium content of fermented lima beans. There

was a significant (Pc0.05) decrease in the iron content of both fermented and sprouted steamed

millet. In contrast, there was significant (Pc0.05) incrcase in the iron contcnt of both fcrmcnted

and sprouted steamed pigeon pea. Similar increase has been made in germinated fenugrcck seed

by El Mahdy el al., (1982) and fermented soybeans by Van der Riet et a1.,(1987). Zinc contcnt of

sprouted and fermented millet and pigeon pea was significantly (P<0.05) increased by steaming.

This increase could probably be due to the removal of antinutrients that might have formed

complexes with zinc. Obizoba and Atii (1994) reported a similar incrcase in the zinc content of

cooked fermented and sprouted millet.

'Table 19. Effect of steaming on antinutritional content of fermented and

sprouted pigeon pea and millet

Samples Tannin Phvtate Cvanide

I [MF 2.80 0.01 17 4.9

SSMF 2.84 0.0075 4.9

FSMF 3.20 0.0030 4.5

(JPF 2.70 0.1 177 4.7 .

SSPF 4.62 0.0150 4.0

FSPF 4.50 0.0250 3.8

LSD (0.05) 0.546 0.00593 0.42

Mean values of triplicate determinations

I JMF - Untreated Millet Flour

SSMF - Sprouted Steamed Millet Flour

FSMF - Fermented Steamed Millet Flour

(JPF - Untreated Pigeon pea Flour

SSPF - Sprouted Steamed Pigeon pea Flour

FSPF - Fermented Steamed Pigeon pea Flour

4.1.7 Effect of steaming on antinutritional content of fermented and

sprouted pigeon pea and millet.

The antinutrient content of fermented steamed and sprouted steamed millet and pigeon

pea is shown in Table 19. The tannin levels of fermented steamed and sprouted steamed millet

were 3.20mglg and 2.84mglg respectively. Furthermore, steaming had no significant (P0.05)

cffcct on thc tannin Ievcl of treated millet. The tannin lcvcls of fcrmcntcd steamed and sproutcd

steamed pigeon pea were 4.50mglg and 4.62mglg rcspectively, Howcvcr, steaming significantly

(PK0.05) increased the tannins levels of fermented and spouted pigeon by 66.67% and 71.1 1%

respectively. The observed increase in the tannin level could be attributed to the increased

formation of tannin -protein complexes by free tannins which was leached out, resulting in

decreased protein value and increased tannins level. The phytate level of the untreated pigeon

pea (0.1 18mg/g) was significantly (PK0.05) higher than the phytate level of the fermented 9

steamed (0.025mglg) and sprouted steamed (0.015mg'/g) pigeon pea. Steaming caused a

significant (P<O.O5) reduction in the phytate level of the fermented steamed millet (0.003mglg)

and sprouted steamed millet (0.0075mg/g), when compared to the untreated millet (0.012mg/g).

This reduction in the phytate level was due to heat treatment and the activity of the enzynics

which brokedown phytate-complexes to release phytate. The phpate was then leached out. 'l'he

cyanide levels of the untreated pigeon pea was 4.7mg/g and fbr the fermented steamed pigcon

pea was 3.8mg/g while for the sprouted steamed pigeon pea, it was 4.0mg/g. The cyanide level

of the untreated millet was 4.9mg/g and the fermented steamed millet was 4.5mdg while

sprouted steamed millet was 4.9mglg. From Table 19, it was evident that steaming significantly

(P<0.05) reduced the cyanide level of treated pigeon pea but not the cyanide level of treated

millet. This could be attributed to the fact that heat removes most HCN (Obizoba and Atii,

1994).

4.2 Functional Properties

4.2.1 Effect of Fermentation on Functional Properties of Millet and

Pigeon pea Flour

The functional properties of unfermented and fermented pigeon pea and millet flours are

shown in Table 20.

Viscosity

The viscosity of 6% concentration of the unfermented and fermented pipcon pea and

millet flours are presented in Table 20. The viscosity of the unfermented millet was 238cp while

that of the fermented millet ranged from 214cp(24hr), 218cp(48hr), 225cp(72hr) to 227.5(96hr). /

There was a significant (P<0.05) decrease in the viscosity of fermented millet. This reduction is /

i

probably due to the activities of amylase that break starch down into simpler sugars, thus \

reducing viscosity (Mensah el al., 1991). This reduction in viscosity duc to fermentation would \

produce thin porridges, which would be good for infant formula. A similar reduction in viscosity

of fermented maize was observed by Mensah et al., (1991). The viscosity of unfermented pigeon

pea was 233cp while the fermented samples showed viscosities of 225cp(24hr), 228cp(48hr),

236.0cp(72hr) and 229.0cp(96hr). From the Table, it could be

Table 20: E

ffect of fermentation on functional properties of m

illet and pipeon pea flours

Functional P

roperties

Samples

Viscosity

Water A

bsorution W

ater Solubilitv L

east Gelation

Bulk D

ensity R

econstitution icp)

Index (mug)

Index (%)

Concentration i%

) id

ml)

Tim

eis)

UM

F 238.0

1.65 288.70

4.0 0.768

75.0

UPF

233.0 1.42

LSD

(0.05) 11.16

0.264 1 1.393

1.20 0.0396

6.09

Mean

values of triplicate determinations

UM

F -Unferm

ented M

illet Flour U

PF - U

nfermented Pigeon pea Flour

FMF24 -

24hr Fermented M

illet Flour FPF24 -

24hr Fermented Pigeon pea Flour

FMF48 -

48hr Fermented M

illet Flour

FMF72 -

72hr Fermented M

illet Flour

FPF48 - 48hr Ferm

ented Pigeon pea Flour

FPF72 - 72hr Ferm

ented Pigeon pea Flour

FMFg6 -

96hr Fermented m

illet Flour FPFg6 -

96hr Fermented Pigeon pea Flour

deduced that fermentation showed no significant effect on the viscosity of the fermented pigeon

pea.

Water Absorption Index:

The water absorption index of the fermented millet compared with that of the

unfermented millet. Fermentation had no significant effect on the water absorption index of the

fermented millet. The water absorption index of the fermented pigeon pea ranged from 1.741nllg

to 2.19mllg. In contrast, fermentation caused a significant increase (P<0.05) in the water

absorption index of fermented pigeon pea. The increase was attributed to the fact that

fermentation enhanced the hydrolysis of starch, which invariably increased the water absorption

index of the samples (Onwulata et al., 1998).

Water Solubility Index:

The water solubility index of the fermented millet samples were 325.9% (24hr), 326.5%

(48hr), 328.9% (72hr) and 331.9% (96hr) respectively. The unfermented millet had a water

solubility value of 288.7%. Fermentation significantly (P<0.05) increased the water solubility

index of millet. The increase in water solubility index was attributed to possible

depolymerisation of the inherent starch and hence to a reduction in the molecular length of

amylose and amylopectin chains, giving rise to the observed incrcase in solubility (Onwulata et

al., 1998). The water solubility index of unfermented pigeon pea was 361.55%. Unlike millet,

there was a significant (P<0.05) decrease in the water solubility index of fermented pigeon pea.

The water solubility index of fermented pigeon pea ranged from 228.8%(24hr), 281.5%(48hr),

28 1.3%(72hr) to 28 1.5%(96hr) (Fig 4).

Least Gelation Concentration :

The least gelation concentration was determined as that concentration of a sample, which

did not fall down or slip on inversion cif the test tubes. The unfermented millet had a comparable

value of 4% with the fermented millet. This shows that fermentation had no significant (P0.05)

effect on the ability of millet to form stable gel. Fermentation of pigeon pea at different hours

caused a change in the least gelation concentration. The least gelation concentration was

observed to be same for both the unfermented and 24hr fermented pigeon pea (4%) but increased

significantly (W0.05) from this 4% to 6% (48hr), to 8% (72hr) and to 10% (96hr). This increase

in the least gelation concentration of fermented pigeon pea suggests that there is a decreased

ability of the fermented pigeon pea flour to form a stable gel. This observation was also noted by

Ihekoronye (1 986).

Bulk Density:

The bulk density of the unfermented millet was 0.768g/ml while that of the fermented t

millet were 0.802g/ml (24hr), 0.801g/ml (48hr), 0.796g/ml (72hr) and 0.832g/m1 (96hr). The

bulk density of the unfermented pigeon pea was 0.877g/ml while that of the fermented pigeon

pea were 0.977g/ml (24hr), 0.947glml (48hr), 0.915g/ml (72hr) and 0.914g/ml (96hr). It was

observed that the bulk density of the unfermented pigeon pea and millet did not differ

significantly from that of the fermented pigeon pea and millet. Therefore, the comparable values

of the bulk density of both unfermented and fermented pigeon pea and millet could be attributed

probably to treatment.

Reconstitution Time:

All the samples reconstituted well in water. The rate of reconstitution of the unfermented

samples (pigeon pea and millet) was higher (66s and 75s respectively) than that of the fermented

samples. There was significant (P<0.05) difference in the reconstitutability of the fermented

sample compared to the reconstitutability of the unfermented samples, but there was no

significant difference between the reconstitution values of the fermented samples.

4.2.2 Effect of Sprouting on Functional properties of Millet and Pigeon

pea Flour

The effect of sprouting on the functional properties of pigeon pea and millet is shown in

Table 2 1.

Viscosity: , \ \

The viscosity of 6% concentration of sprouted millet varied from 221 Scp (24hr), 2 2 8 . 5 ~ ~

(48hr), 2 2 4 . 5 ~ ~ (72hr) to 2 2 5 . 5 ~ ~ (96hr), Sprouting significantly (P<0.05) reduced the

viscosity of millet flours. The reduction in viscosity in sprouted millet flours was as a result of

starch degradation caused by the action of alpha - and beta - amylases, that developed during the

germination process (Mosha and Svanberg, 1983). Marero et al., (1988) reported similar

decrease in viscosity of rice and corn due to germination. In contrast, sprouting of pigeon pea did

not show any such decrease in apparent viscosity except for the 96hr-sprouted pigeon pea that 1

Table 21: E

ffect of sprouting on the functional properties of sprouted millet and pigeon pea flours

Functional P

roperties

Samples

Viscositv

Water A

bsomtion

Water

Solubility L

east Gelation

Bulk D

ensity R

econstitution (C

D)

Index (mug)

Index (Oh)

Concentration (%

) (dm

11 T

ime(s1

UM

F 238.0

1.65 288.70

4.0 0.768

75.0

sm

48

228.5 1.68

293.20 4.0

0.676 5 1 .O

SMF72

224.5 1.55

205.00 4.0

0.649 44.7

sm96

225.5 2.78

21Q.00

4.0 0.659

57.0

UPF

233.0 1.42

361.55 4.0

0.877 66.0

SPF24 233.5

1.78 376.70

6.0 0.943

63 .O

SPF48 ,

233.5 1.83

307.00 6.0

0.975 57.0

SPF72 233.0

1.99 313.70

8.0 0.987

51.0

SPF96 212.0

2.05 277.60

6.0 0.975

54.0

LSD

(0.051 7.48

0.298 4.299

1.42 0.0536

5.55

Mean

values of triplicate determinations

UM

F -Unsprouted M

illet Flour U

PF - U

nsprouted Pigeon pea Flour

SMF24 -

24hr Sprouted Millet Flour

SPF24 - 24hr Sprouted Pigeon pea Flour

SMF48 - 48hr Sprouted M

illet Flour SPF48 -

48hr Sprouted Pigeon pea Flour

SMF12 -

72hr Sprouted Millet Flour

SPF12 - 72hr Sprouted Pigeon pea Flour

SMF96 -

96hr Sprouted Millet Flour

SPFg6 - 96hr Sprouted Pigeon pea Flour

exhibited reduced (I+-0.05) viscosity. However, this result agrees with the result of Marero et ul.,

(1 988), who observed a reduction in the viscosity of germinated mungbean and cowpea after

48hr.

Water Absorption Index: ,

The water absorption index of sprouted millet ranged from 1.55mllg to 2.78mllg while

that of unsprouted millet was 1.65mllg. Also, the water absorption index of sprouted pigeon pea

ranged from 1.78mVg to 2.05mllg while that of the unsprouted pigeon pea was 1.42mllg. The

water absorption index of sprouted pigeon pea increased as the hours of sprouting increased.

However, sprouting showed no significant (P>0.05) effect on the water absorption index of

millet except for 96hr-sprouted millet in which there was significant (Pc0.05) increase.

Furthermore, sprouting significantly (P<0.05) increased the water absorption index of pigeon

pea. Onimawo and Asugo (2004) reported a similar increase in the water absorption index of

pigeon pea due to germination. The high water absorption index could be attributed to either high

protein content or more of hydrophilic polysaccharides during the course of sprouting.

Furthermore, the high water absorption index indicates that the sprouted seed flour could be

useful in food systems, which require hydration to improve handling characteristics and to

maintain freshness.

Water Solubility Index:

The water solubility index for sprouted millet was varied. The water solubility index of

unsprouted millet was 288.7%while that of the sprouted millet varied from 315.6% (24hr),

293.2% (48hr), 205.0% (72hr) to 210.0% (96hr). Sprouting significantly (P<0.05)

increased the water solubility index of 24hr and 48hr sprouted millet. However, a significant

(P<0.05) decrease was observed as sprouting time extended beyond 48hr. This decrease could be

as a result of degradation of starch during sprouting. Gujska and Khan (1991), observed similar i \

\ decrease in water solubility index of extruded pinto bean flour. Furthermore, the water solubility

index of pigeon pea showed a similar trend as that of millet. The water solubility index of the

unsprouted pigeon pea was 361.5394, which was significantly (Pr-0.05) increased at 24hr

sprouting and this could as well be attributed to starch degradation during sprouting.

Least Gelation Concentration:

Gel formation is primarily related to the concentration of amylose in the flour starch.

Sprouting did not cause any significant change in the least gelation concentration of millet. The

least gelation concentration of unsprouted and sprouted millet was 4%. Sprouting caused an t

increase in the least gelation concentration of pigeon pea. Unsprouted pigeon pea had a least

gelation concentration of 4% while the least gelation concentration of sprouted pigeon pea

increased to 8%. The increase in the least gelation concentration implies a decreased ability of

the pigeon pea flour to form stable gels. Onimawo and Asugo (2004), reported a similar increase

in the least gelation concentration of germinated pigeon pea of 10%. However, concluded that

the variation in the gelling properties of different legume flours in associated with the relative

ratios of different constituents such as protein, carbohydrate and lipids.

Bulk Density:

Bulk density is a reflection of the load the sample can carry if allowed to rest directly on

another. The bulk density values of both sprouted millet and pigeon pea is shown in Table 21.

The bulk density of the unsprouted millet flour was 0.768gml. Sprouting caused a significant

decrease (P<0.05) in the bulk density of millet except for millet sprouted for 24hr which had a

comparable value of 0.760glml. The decrease in bulk density could be attributed to the

processing method of sprouting. Balandran - Quintana et a1.,(1998) reported a decrease in the

bulk density of extruded whole pinto bean. However, sprouting significantly (P<0.05) increased

the bulk density of pigeon pea, when compared to the unsprouted pigeon pea, whose bulk density

value was 0.877glml. Onimawo and Asugo (2004), reported similar observation of increase in

bulk density of germinated pigeon pea, while, on the contrary, Akubor and Chukwu (1999),

observed a decrease in bulk density in African oil bean when fermented.

Reconstitution Time: '. The reconstitution time for both sprouted pigeon pea and millet reduced when compared

to the unsprouted pigeon pea and millet. It could therefore, be said that sprouted samples

reconstituted well in water.

Table 22: E

ffect of steaming on functional properties of ferm

ented steamed and sprouted steam

ed millet and

pipeon pea flours

Functional P

ro~erties

Sa

m~

les V

iscosity W

ater ~b

sorption

W

ater Solubilitv L

east Gelation

Bulk D

ensitv R

econstitution (C

D)

Index (mug)

Index (%)

Concentration (%

) (d

ml)

Tirne(s1

UM

F 238.0

SSMF

220.0

FSMF

215.0

UPF

233.0

SSPF 228.0

FSPF 223 .O

1.65 3 17.40

10.0 0.789

60.0

LSD

(0.05) 8.95

0.299 4.272

2.62 0.0238

4.42

Mean values of triplicate determ

inations

UM

F -Untreated

Millet Flour

UPF -

Untreated Pigeon pea Flour

SSMF -

Sprouted Steamed M

illet Flour

FSMF -

Fermented Steam

ed Millet Flour

SSPF -Sprouted Steamed Pigeon pea Flour

FSPF -Fermented Steam

ed Pigeon pea Flour

4.2.3 Effect of steaming on functional properties of fermented and

sprouted pigeon pea and millet flour:

Table 22 shows the hnctional properties of fermented steamed and sprouted steamed

pigeon pea and millet flours.

Viscosity:

Result of the viscosity measurements of the fermented steamed and sprouted steamed

pigeon pea and millet is shown in Table 22. The viscosity of the.untreated millet was 238cp

while that of untreated pigeon pea was 233cp. The viscosities of the fermented steamed and

sprouted steamed millet were 215cp and 220cp respectively, while the viscosities of the

fermented steamed and sprouted steamed pigeon pea were 223cp and 228cp respectively. The

result showed that steaming significantly (PC0.05) reduced the viscosity of fermented and

sprouted millet and pigeon pea. Mercier et al., (1975) observed similar decrease in viscosity of

corn and rice due to extrusion cooking. The reduction in viscosity could be attributed to the

degradation of starch due to steaming.

Water Absorption Index:

Among the fhctional properties, water absorption index is irnportant because of the

hydrogen bonds formed between water and polar residues of protein molecules. Significant

increases (P<0.05) were observed in water absorption index of fermented steamed and sprouted

steamed millet and pigeon pea. Balandra-Quintana et a1.(1998) observed a similar increase in

water absorption index of extruded pinto bean. The increase was attributed to the increase in

temperature, which caused amylose and amylopectin separation, forming an expansible matrix,

which result in a higher water absorption index.

Water Solubility Index:

The water solubility index for the untreated millet was 288.7% while for the ferrnentcd

steamed and sprouted steamed millet was 368.2% and 366.5% respectively. Significant (P<0.05)

increase was observed in the water solubility index of fermented steamed and sprouted steamed

millet. This increase could be attributed to starch depolymerization at higher temperaturcs,

reducing molecular length of amylose and amylopectin chains. This result confirmed those of

Anderson (1982) who had extruded corn and sorghum. The water solubility index of untreated

pigeon pea was 361.55%. Steaming caused a significant increase (K0.05) in the water solubility r

index of sprouted pigeon pea. However, a significant decrease (K0.05) was observed in the

water solubility of fermented steamed pigeon pea. The observed reduction in the water solubility

index of fermented steamed pigeon pea, could probably be attributed to the denaturation and .

aggregation of protein due to steaming. Gujska and Khan (1991) observed a similar decrease in

water solubility index of extruded pinto bean flour. On the other hand, the observed increase in

water solubility index for sprouted steamed pigeon pea, could probably be attributed to minimal

denaturation of protein during processing. Balandran- Quintana el al. (1998), observed similar

increase in water solubility index for extruded whole pinto bean.

Least Gela tion Concentration:

The effect of steaming on the least gelation concentration of untreated, sprouted and

fermented millet and pigeon pea flours is shown in Table 22. The least gelation concentration of

untreated pigeon pea and millet was 4%. Steaming caused a significant (Pc0.05) increase in the

least gelation concentration of sprouted and fermented millet and pigeon pea, when compared to

the untreated millet and pigeon pea. Least gelation concentration was observed to increase from

4% for the untreated pigeon pea to 10% for both fermented and sprouted pigeon pea. It was also

observed to increase from 4% for the untreated millet to 10% for sprouted millet and 8% for

fermented millet. However, the increase in least gelation concentration by steaming could

probably lead to a decrease in the ability of samples to form gel. This decease in ability to form

gel, could be attributed to heat denaturation.

Bulk Density:

Steaming caused a decrease in the bulk density of both sprouted and fermented pigeon

pea. The bulk density of untreated pigeon pea was 0.877gIrnl while for the sprouted steamed

pigeon pea it was 0.794g/rnl and 0.789g/ml for fermented steamed pigeon pea. This decrease

could be due to starch degradation resulting in less expansion due to high temperature effect.

Steaming caused a decrease in the bulk density of fermented steamed millet, whose value was

0.735gIml but an increase in the bulk density of the sprouted steamed millet, 0.777g/ml when

compared to the untreated millet, 0.768glml.

Reconstitution Time:

All samples reconstituted well in water. There was a reduction in the reconstitution time '

of the fermented steamed and sprouted steamed millet and pigeon pea when compared to the

, untreated samples.

4.3 Sensory Evaluation:

The mean sensory scores of the infant food formulations, casein and commercial diet are

shown in Table 23.

Colour:

Colour scores ranged fiom 6.8 to 5.1. The commercial diet had colour score of 6.8 which

was significantly (Pc0.05) higher than those of the formulated and casein diets. Casein diet had

comparable (P>0.05) colour score of 5.8 with diets B (untreated pigeon pea flour + fermented

steamed millet flour) and C (untreated pigeon pea flour + sprouted steamed millet flour) (5.4) but

significantly (P<0.05) differed fiom the rest. Furthermore, among the formulated diets, there was

no significant (P>0.05) difference in terms of colour score.

Flavour:

Flavour scores ranged fiom 5.9 to 4.0. The commercial diet had the highest flavour score

of 5.9 while diet B (untreated pigeon pea flour + fermented steamed millet flour) had the lowest

flavour score of 4.0. However, the flavour score of the commercial diet did not differ (P>0.05)

fiom the flavour score of diets H (sprouted steamed pigeon pea flour + fermented steamed millet

flour) (5.8), F (fermented steamed pigeon pea flour + sprouted steamed millet flour) (5.7), I

(sprouted steamed pigeon pea flour + sprouted steamed millet flour) (5.9, E (fermented steamed

pigeon pea flour + fermented steamed millet flour) (5.2) and casein (5.6).

Consistency: The commercial diet had significantly (P<0.05) higher consistency score compared to the

formulated and casein diets. On the other hand, casein diet had the lowest consistency score of

3.7. Diets A(4.4), B(4.4), C(4.1) and D (4.0) had comparable consistency score with casein.

Furthermore, diets E(5.5), F(5.2) and l(5.3) had comparable consistency score.

Reconstitution Time:

All samples reconstituted well in water. There was a reduction in the reconstitution time

of the fermented steamed and sprouted steamed millet and pigeon pea when compared to the

, untreated samples.

4.3 Sensory Evaluation:

The mean sensory scores of the infant food formulations, casein and commercial diet are

shown in Table 23.

Colour:

Colour scores ranged fiom 6.8 to 5.1. The commercial diet had colour score of 6.8 which

was significantly (P<0.05) higher than those of the formulated and casein diets. Casein diet had

comparable (P>0.05) colour score of 5.8 with diets B (untreated pigeon pea flour + fermented

steamed millet flour) and C (untreated pigeon pea flour + sprouted steamed millet flour) (5.4) but

significantly (P<0.05) differed fiom the rest. Furthermore, among the formulated diets, there was

no significant (P>0.05) difference in terms of colour score.

Flavour:

Flavour scores ranged fiom 5.9 to 4.0. The commercial diet had the highest flavour score

of 5.9 while diet B (untreated pigeon pea flour + fermented steamed millet flour) had the lowest

flavour score of 4.0. However, the flavour score of the commercial diet did not differ (P>0.05)

fiom the flavour score of diets H (sprouted steamed pigeon pea flour + fermented steamed millet

flour) (5.8), F (fermented steamed pigeon pea flour + sprouted steamed millet flour) (5.7), I

(sprouted steamed pigeon pea flour + sprouted steamed millet flour) (5.9, E (fermented steamed

pigeon pea flour + fermented steamed millet flour) (5.2) and casein (5.6).

Consistency: The commercial diet had significantly (Pc0.05) higher consistency score compared to the

formulated and casein diets. On the other hand, casein diet had the lowest consistency score of

3.7. Diets A(4.4), B(4.4), C(4.1) and D (4.0) had comparable consistency score with casein.

Furthermore, diets E(5.9, F(5.2) and l(5.3) had comparable consistency score.

Table 23: Sensory evaluation scores of the formulated products, commercial

diet and casein diet

Quality Attributes

Samples Colour Flavour Consistency Mouth Taste After- Overall

feel taste Acceptability

A 5.2 5.0 4.4 3.9 4.3 3.8 4.5

B 5.4 4.0 4.4 3.8 4.2 3.6 4.6

C 5.4 4.9 4.1 3.8 4.9 3.9 4.7

D 5.3 4.9 4.0 3.9 4.9 3.8 5.0

E 5.1 5.2 5.5 3.5 5.9 3.7 5.6

F 5.3 5.7 5.2 4.0 5.9 3.6 5.7

G 5.2 5.1 4.6 4.1 4.0 4.1 4.8

H 5.3 5.8 5.8 3.7 5.5 3.6 5.4

I 5.1 5.5 5.3 3.8 6.0 3.9 5.5

J 6.8 5.9 6.6 6.4 6.4 4.3 6.8

K 5.8 5.6 3.7 5.7 6.0 3.9 5.8

LSD (0.05) 0.42 0.73 0.76 0.79 0.62 0.92 0.72

Mean of 30 replicates

A-UPF+UMF . J - Commercial Diet

B - UPF + FSMF K - Casein Diet

C - UPF + SSMF UPF -Untreated Pigeon pea Flour

D - FSPF + UMF UMF - Untreated Millet Flour

E - FSPF + FSMF FSMF - Fermented Steamed Millet Flour

F - FSPF +-SSMF SSMF - Sprouted Steamed Millet Flour

G - SSPF + UMF

H - SSPF -t FSMF

I - SSPF + SSMF

SSPF - Sprouted Steamed Pigeon pea Flour

FSPF - Fermented Steamed Pigeon pea Flour

Mouth feel:

Commercial diet had comparable (P>0.05) mouth feel score of 6.4 with those of casein

diet (5.7). On the contrary, commercial diet had significantly (W0.05) highcr mouth fccl score

than the formulated diets. Furthermore, there was no significant (PB0.05) difference among the

mouth feel scores of formulated diets.

Taste:

The taste score ranged from 6.4 to 4.0. Commercial diet had the highest taste score of 6.4.

Furthermore, commercial diet had comparable (P>0.05) taste score with casein (6.0), diets I(6.0),

E(5.9) and F(5.9) but was significantly (P<0.05) higher than those of other formulated diets. Diet

G(4.0) had the lowest taste score which was comparable to the taste. score of diets A(4.3) and

B(4.2).

Aftertaste

The aftertaste score ranged from 4.3 to 3.6. Commercial diet had the highest aftertaste

score (4.3) while diets B, F and H had the lowest level of 3.6. There was no significant (PB0.05)

difference in the aftertaste score of commercial, casein and formulated diets.

Overall Acceptability:

The overall acceptability of the diets were influenced by the organoleptic attributes of

colour, flavour, taste, mouthfeel etc. The overall acceptability score ranged from 6.8 to 4.5.

Commercial diet had the highest overall acceptability score of 6.8. There was significant

(P<0.05) difference in the overall acceptability score of the commercial diet to those of casein

(5.8) and formulated diets. On the contrary, there was no significant (P>0.05) difference in the

overall acceptability score between the casein diet (5.8) and those of diets F (5.7), E (5.6), I(5.5)

and H (5.4). Diet A had the lowest overall acceptability score of 4.5, which was comparable to

the overall acceptability scores of diets B(4.6), C(4.7), and D(5.0).

The preference for the commercial diet was not surprising. The panelists were familiar

with the organoleptic properties of gruels from commercial diet (Cerelac) which is popularly

used as complementary food in Nigeria, and is vade from only maize. Promotion will be \

\

required to popularize the composites from other sources of cereals and legumes which have a

lot of nutritional benefits over cereal complementary diet alone (Nnarn, 2001). Some of the

organoleptic properties of the infant food formulation can be improved by addition of edible

flavouring and colouring materials (Nwanekezi, 200 1).

Based on the organoleptic qualities-colour, flavour, mouthfeel and overall acceptability

of diets studied, samples A for negative control, E, F, H, I, J and K, were chosen for fkrther w

studies. These samples were analysed for hnctional properties and subsequently used for animal

bioassay. For the animal bioassay diets A, E, F, H, 1, J and K were designated, 1, 11, 111, 1V, V, VI

and VII respectively.

4.4 Animal Bioassay

4.4.1 Proximate Composition of Diets used for Animal Bioassay Table 24 shows the proximate composition of blends from pigeon yea and millet. The

proximate composition of the blends were compared with casein and a commercial diet. The

protein values for the different diets were diet I, 9.43%, diet 11, 9.43%, diet 111, 8.8%, diet IV,

8.70% and diet V, 10.50%. The protein content of diet I and I1 were comparable (DO.05) to the

protein content of the casein diet, 9.53% which is comparable to the FA0 standard protein value

of 10% (FAO, 1994). Furthermore, the protein content of diet V was significantly (K0.05)

higher than those of casein. It could be seen that the use of composite flours could serve as a

practical means of upgrading protein levels of millet flour, as the protein level of the composite

flours were influenced by supplementation (Nnam, 2001). Commercial diet had significantly

(P<0.05) higher protein value of 19.53%, than those of the composite, but it was con~parable to

the 15% FAO/WHO requirement for commercial diet. The casein diet had comparable (PB0.05)

lipid content (1 1.43%) with diets I (1 1.13%), IV (1 1.33%) and V (1 1.73%), which were

significantly (P<0.05) higher than the lipid value (8.4%) of commercial diet. Furthermore, diet I1

and 111 had significantly (W0.05) lower lipid content than those of casein diet. Casein diet had a

crude fibre content of 2.7% which is comparable (P0.05) to the crude fibre content of diet V,

but differs significantly (P<0.05) from those of other composite and commercial diets. Similarly,

commercial diet had comparable (P30.05) crude fibre content (2.47%) with diets I and 111. The

moisture content of the composite flour varied. Casein diet had comparable (P>0.05) moisture

content (1.5%) with those of diets 11, IV and V. On the contrary, commercial diet had

significantly (P<0.05) higher moisture content (7.63%), which was comparable to those of diet I

(7.27%). Commercial diet had a comparable (P>0.05) ash content (5.00%) with those of diet IV,

but was significantly (P<0.05) higher than that of casein. Similarly, casein diet had a comparable

(P>0.05) ash content of 3.07% with those of diet V (3.4%). Commercial diet had significantly

(P<0.05) lower carbohydrate (56.96%) and energy (329.2kcaI/lOOg) levels than those of the

Table 24: P

roximate com

position of diets used for animal bioassay (%

dry weight basis)

Samples

Crude protein

Crude fat

Crude fibre

Moisture content

Ash

Nitrogen F

ree Extract

Enerw

T

rue motein

I 9.43

11.13 2.47

7.27 4.10

65.60 400.29

6.70

I1 9.43

9.80 2.40

1 SO

2.60

74.27 423 .OO

5.10

I11 8.80

9.67 2.47

4.35 2.60

72.1 1 4 10.67

5.37

n(r 8.70

11.33 2.50

1 SO

4.80

71.17 42 1.45

4.90

V

10.50 1 1.73

2.70 1 S

O

3.40 70.17

428.25 7.07

VI

19.53 8.40

2.47 7.63

5.00 56.97

379.20 10.40

VII

9.53 1 1.43

2.70 1 S

O

3.07 71.77

428.07 7.30

LSD

(0.05) 0.443

0.463 0.064

0.645 0.449

1.697 4.282

0.572

Mean of 30 replicate determ

inations

I - U

PF + UM

F U

MF -

Untreated M

illet Flour

I1 - FSPF + FSM

F U

PF -Untreated Pigeon pea Flour

I11 -FSPF + SSMF

FSPF - Ferm

ented Steamed Pigeon pea Flour

IV- SSPF + FSM

F SSPF -

Sprouted Steamed Pigeon pea Flour

V-

SSPF + SSMF

FSMF -

Fermented Steam

ed Millet Flour

VI -

Com

mercial D

iet SSM

F - Sprouted Steam

ed Millet Flour

VII -

Casein D

iet

composite flours. Unlike commercial diet, casein diet had a comparable carbohydrate content

(71.77%) with diet 111 (72.1 I%), diet 1V (71.17%) and diet V (70.17%). Casein diet had high

energy level that was comparable (P0.05) to diet V. Similar to the crude protein, the true

protein content of commercial diet was significantly higher (P<0.05) than those of the composite

flours and casein

4.4.2 Mineral Composition of diets used for animal bioassay

The mineral content of the composite flour is shown in Table 25. There was a variation in

the calcium content of composite flours with diet V having the lowest level. The commercial diet

(4.58mglg) had comparable (P>0.05) calcium content with casein diet (4.63mdg). On the

contrary, diets 1(6.63mg/g), III(5.07mg/g), IV(4.97mgk) and 11(4.80mg/g) had significantly

(P<0.05) higher calcium content than those of casein and commercial diets. Commercial diet had

iron content (6.98mglg) that is significantly (PC0.05) higher than that of casein diet(4.76mdg).

Diet II(7.10mglg) had the highest iron content that was comparable (P>0.05) to those of

commercial diet and diet 111. However, casein diet had a low iron content that was comparable to

diet V (4.68mg/g). The composite diet had significantly (K0.05) higher iron content than those

of casein except for diet V. This could be compared to the work by Nkama and Malleshi (1998)

in which iron content was observed to be high in the diet formulated with rice, pearl millet,

cowpea and groundnut. The zinc content of composite flours ranged from 46.30mg/g (diet I), to . 45.67mdg (diet III), to 45.00mg/g (diet V), to 43.33mg/g (diet 1V) to 29.00mglg (diet 11). Diets

I, 111 and V had comparable zinc content which were significantly (Pc0.05) higher than those of

casein and commercial diets (41.33mg/g and 40.0mglg respectively). Diet I1 had the lowest zinc

content, though it had the highest iron content. The increase in the zinc content of the formulated

diet could be compared to the increase observed by Nkama and Malleshi (1998) in zinc content I, \

of formulated rice-cowpea and rice -cowpea - groundnut.

Table 25: Mineral Composition of diets used for animal bioassay

Samples Calcium (mglpr) Iron (mglg) Zinc (mglg)

1 6.63 6.66 46.30

I1 4.80 7.10 29.00

111 5.07 6.98 45.67

I V 4.97 5.00 43.33

V 1.03 4.68 45.00

VI 4.58 6.98 40.00

VII 4.63 4.76 41.33

LSD (0.05) 0.189 0.3 16 1.610

Mean values of triplicate determinations

I - UMF + UPF UMF - Untreated Millet Flour

I1 - FSPF + FSMF UPF - Untreated Pigeon pea Flour

111 - FSPF + SSMF FSPF - Fermented Steamed Pigeon pea Flour

IV - SSPF + FSMF SSPF - Sprouted Steamed Pigeon pea Flour

V - SSPF + SSMF FSMF - Fermented Steamed Millet Flour

VII - Commercial Diet SSMF - Sprouted Steamed Millet Flour

VII - Casein Diet

4.4.3 Functional Properties of Diets

The hnetional properties of diets fed to the animals are shown in Table 26.

Viscosity I /

The viscosity measurements of the different composite flours varied. Commercial diet

had the highest viscosity value of 303cp while casein diet had the lowest viscosity value of I,

\

190cp. The viscosities of the diets were significantly (P<0.05) lower than those of the

commercial diet and they ranged fiom 201cp (diet IV), 205cp (diet I), 207cp (diet II), 209cp (diet

V) and 210cp (diet 111). The viscosity of the composite blends was higher than casein diet but

lower than the commercial diet. Diet IV had the lowest viscosity amongst the composite blends.

The viscosity of the composite blends (Table 26) was lower (P<-0.05) when compared to the

viscosity of the fermented steamed and sprouted steamed flours. Nnam (2001) observed similar *

lower viscosity in porridges f?om composite flours when compared to the control, fermented

sorghum flour. This low viscosity is a significant property in weaning formula because ,it

facilitates easier consumption, digestion and greater nutrient intake.

Water Absorption Index: The ability to absorb water by the blends varied. The water absorption index of

commercial diet was low, 1.40mVg. Diet I11 had a high water absorption index of 1.83 mllg. Diet

I, 11, IV and V had a comparable water absorption index with casein diet of 1.57mllg. The water

absorption ability of the fermented steamed and sprouted steamed pigeon pea and millet flours

(Table 22) was higher than the water absorption ability of the composite blends (Table 26). This

reduction in the water absorption index of diets could be attributed to the complementing effects

of the proteins in the diets.

Water Solubility Index:

Commercial diet had significantly (P<0.05) higher water solubility index of 428.0% than

those of casein diet (369.5%). Casein diet had comparable (P>0.05) water solubility index with

diets I and V (370.5% and 376.0% respectively). Diet 11 and IV had comparable water solubility

index which are significantly (P<0.05) lower than those of the casein diet. Diet 111 had the lowest

water solubility index of 306.9%. The reduced water solubility index of formulated diets (JI, 111,

IV) indicates that the diets have easier digestibility when consumed.

Least Gelation Concentration:

The least gelation concentration of composite blend varied. The variation in least gelation

concentration of composite flours was from 4% (diet V), 6% (diet I), 8% (diets 1JI and IV) to

10% (diet 11). Commercial diet had a high least gelation concentration value of 12%. This

implies that its ability to form stable gel is low. Casein did not form gel up to 20%(w/v)

concentration range. This result suggests that gelation is not only the function of quantity of \

protein but also the type of protein(s) and the non-protein components as well.

Bulk Density Determination:

The bulk density of the composite blends varied. Commercial diet had the lowest bulk

density value of 0.641g/ml. Casein diet had comparable bulk density level with diets 111 and V,

0.828g/ml and 0.807dml respectively. Similarly, diet I and IV had comparable bulk density

level. Among the composite blends, diet IT had the lowest bulk density level of 0.773gIm1, but

which was significantly higher than that of the ~ommercial diet.

Reconstitution Time:

All composite blends reconstituted well in water. Comn~ercial diet had the longest

reconstitution time of 78s that was comparable to that of diet I11 (72s). However, diets 1, 1V and

V had lower reconstitution time of 63s, 69s and 69s respectively which compared with the

reconstitution time of casein (66s). Diet I1 had the lowest reconstitution time of 54s. The low

reconstitution time suggests that diet I1 was very dispersible, highly solublc and easily

rehydrated.

4.4.4 Animal Bioassay:

Table 27 shows the result of food intake (g), gain in body weight (g), protein efficiency

ratio (PER), N intake (g), feacal N (g), Urinary N (g), digested N (g), retained N, biological

value (BV) and Net protein utilization (NPU) of rats fed mixed protein diets based on fermented

steamed and sprouted steamed millet and pigeon pea flours. There were differences in food

intake among the groups of rats. The higher food intake of the group of rats fed commercial diet

(173.54g) which was significantly (P<0.05) different from the rest, might bc attributcd to its

palatability. The surprisingly, high food intake (123.428) of group of rats fed diet 1V compared

to those rats fed other mixed protein diets could be attributed to the mutual supplementation

effects of the diet that promoted palatability. This is a common phenomenon as observed by

Obizoba (1 990). The comparable food intake of groups of rat fed diets I11 and I1 as against casein

(1 13.5 lg, 11 1.34g vs 130.798 respectively) might be because of improved flavour or palatability

or both (Obizoba, 1990). The weight gain of rats in various groups differed. The group of rats fed

the commercial diet had the highest weight gain (56.82g), which was significantly higher

(P<0.05) than those groups of rats fed other mixed protein diet. The high maintenance body

weight of group of rats fed commercial diet showed that it contained desirable and adequate

essential amino acid, which these rats used to synthesize new body protein tissue (Ossai and

Malomo, 1988). On the other hand, the comparable value in maintenance body weight of rats fed

diet 111 (13.1 0g) and casein (15.968) indicates that when plant protein are mixed to complement

each other, it is possible to produce adequate protein that will be equal or higher than that of

animal protein (casein) (Ossai and Malomo, 1988). The low maintenance weight gain of groups

of rats fed diet I, could be attributed to poor protein utilization (Obizoba, 1986), or that the

Table 27: F

ood intake, gain in body weipht, protein efficiency ratio (P

ER

), N intake, feacal N

, urinary N,

di~

ested N, retained N

, biological value (BV) and net protein utilization of rats fed diets based on ferm

ented and sprouted m

illet and pipeon pea. D

iets I

I1 I11

IV

V

VI V

II

N ratios

70:30 70:30

70:30 70:30

70:30 100:O

100:O

Food intake (g) 90.89

11 1.34 113.51

123.42 102.08

173.54 130.79

Wt gain (g)

6.98 10.41

13.10 10.25

10.00 56.82

15.96

* PER

0.584

0.712 0.876

0.630 0.748

2.5 0.925

N intake (g)

0.322 0.364

0.372 0.471

0.364 0.756

0.514

Feacal N (g)

0.0182 0.01 19

0.0098 0.0101

0.01 82 0.0289

0.0209

Digested N

(g) 0.3040

0.3525 0.3689

0.4609 0.3458

0.7271 0.493 1

Urinary N

(g) 0.1081

0.1066 0.1080

0.1097 0.1083

0.1383 0.0785

Retained N

(g) 0.1957

0.2455 0.2609

0.3512 0.2375

0.5888 0.4146

BV

64.42

69.73 70.72

76.20 80.98

84.08 68.68

NPU

60.77

67.45 70.13

74.57 65.25

77.88 80.66

*PE

R converted to that of casein standard (2.5)

Means of 35 replicates

I - U

MF + U

PF U

MF -

Untreated M

illet Flour

I1 - FSPF + FSM

F U

PF -Untreated Pigeon pea Flour

I11 -FSPF + SSMF

FSPF - Ferm

ented Steamed Pigeon pea Flour

IV- SSPF + FSMF

FSMF -

Fermented Steam

ed Millet Flour

V-

SSPF + SSMF

SSPF - Sprouted Steam

ed Pigeon pea Flour

VI - C

omm

ercial Diet

SSMF -

Sprouted Steamed M

illet Flour

VII -

Casein D

iet

protein is deficient in one or more essential amino acids (Ossai and Malomo, 1988), or due to

type of legume and differences in essential amino acid content of proteins (Obizoba, 1990). The '

higher food intake of diet IV as against that of diet V and their comparable maintenance body

weight indicate that both are equally good to maintain body N, even though diet IV had higher

food intake, and vice versa. The PER (protein efficiency ratio) values were converted to a value

of 2.5 for casein. There were variations in PER for all groups, which was an indication that PER

is influenced by weight gain and source of N (Obizoba, 1990). The higher (P<0.05) PER for the

group of rats fed commercial diet (2.5) might he associated with the higher food intake as well as

the maintenance body weight. The comparable PER value for groups of rats fed diet I11 and

casein (0.876 and 0.925 respectively), confirms the observation made over !he maintenance body

weight. The comparable PER of rats fed diets 11, IV and V (0.712, 0.6:30 and 0.748 respectively)

indicates that the three protein diets had comparable nutritive vidue. This is a common

phenomenon as observed by Okeke and Obizoba (1986). The 7-day N intake for all groups of

rats varied. The N intake of the groups of rats followed a trend towards higher values for those

rats with higher food intake, for example, the commercial diet group of rats that had the highest

food intake (174.54g), maintenance body weight (56.828) and PER (2.5) had the highest N

intake of 0.7568. Feacal N was a hnction of source of N and intake. The group of rats fed casein

diet had high N intake, high feacal N and also high digested N. Feacal N affects digestibility. The

lower feacal excretion of groups of rats favoured their increased N digestibility. The digested N

for all group of rats varied and was a hhction of N intake and feacal output. The group of rats

fed commercial diet had a significantly higher (P<0.05) digested N value (0.72718). which

indicated that it provided the essential amino acid required for high digestibility (Onuoha and /

Obizoba, 2001). The digested N value (0.4931g) of casein which was cdmparable (P>0.05) to

that of diet IV indicated that diet IV provided protein with desirable essential amino acids which \

were equal to that of casein and were equally digestible (Onuoha and Obizoba, 2001). The higher \

digested N value of commercial diet (0.72718) indicated superiority over the lower digested N

values of diets I11 (0.3689g), I1 (0.3525g), V (0.34588) and I (0.304g) (Obizoba,1986). The

urinary N excretion for all groups of rats varied and might be due to biological variation of the

animals and protein quality (Obizoba, 1986). The groups of rats fed commercial diet had high

urinary N excretion of 0.13838, which adversely affected their retained N, biological value (BV)

and net protein utilization (NPU). However, the groups of rats fed commercial diet that had 1.

higher digested N and higher urinary N, also had higher N retention but reduced biological value

(BV) and net protein utilization (NPU), when compared to the group of rats fed casein diet.

Casein diet had high digested N but low urinary N, which resulted in high retained N, biological

value and subsequently net protein utilization (NPU). The retained N of the group of rats fed

formulated diets ranged from 0.1957g to 0.3512g. Diet IV had significantly (P<0.05) higher

retained N (0.35 12g) than those of other formulated diets, which resulted in high biological value

and net protein utilization. The groups of rats fed diets 11, 111 and V had comparable retained N

values of 0.2455g, 0.2609g and 0.2373g respectively. The retained N value, however, was a

hnction of food intake, N intake, feacal N and urinary N. The trend towards high net protein

utilization for group of rats fed diets 111, IV, commercial diet and casein diet suggests that the

protein source was of good quality. Similarly, it suggests that the diet combinations provided a

good pattern of essential amino acids, which the rats used for body tissue synthesis. As judged by

net protein utilization (NPU), diets IV and 111 were superior to others.

CHAPTER FIVE

5.0 Conclusions, Recommendations and Suggestions for Further

Studies

5.1 Conclusions

The result of this study shows the formulation complementary (weaning) formula. The

production of this complementary formula was through simple but adequate processing methods

that can easily be adopted by many families. The findings of this study could be summarized as

follows:

Fermentation increased the protein content of pigeon pea from 22.04% to 23.46% and

millet fiom 9.18% to 9.46%. Sprouting also increased the protein content of pigeon pea from

22.04% to 24.85% but reduced that of millet fiom 9.18% to 7.88%. Fermentation reduced the

antinutrients-tannin (2.7mdg to 2.26mg/g), phytate (0.117mglg to 0.0018mglg) and cyanide

(4.9mg/g to 4.5mgJg) levels of millet and pigeon pea. There was a variation, however, in the

antinutrient level of sprouted samples. Steaming reduced the protein, calcium, phytate and

cyanide content of the fermented and sprouted pigeon pea and millet samples, but increased the

zinc, iron and tannin content. The protein content of the formulated products was comparable to

the protein content of casein diet (9.53%) and the FA0 standard protein value of 10%. Among

the most acceptable formulation (E, F, H and I which were designated as 11, 111, 1V and V

respectively) used for animal assay, group' of rats fed diet lV, had food intake (123.42g), digested

N (0.4609g) and net protein utilization (74.57), which were comparable with those of casein diet

(1 30.7913, 0.493 1g and 80.66 respectively).

It is evident fiom the results that the blending of fermented millet and sprouted pigcon

pea in the proportion used in the present study provides a formulation that compares favourably

with casein.

5.2 Recommendations

From the conclusion made in this research work we recommend the following:

Raw materials for food, especially, weaning food should be fermented or germinated and

steamed before formulation. This increases the nutritive quality.

A 72-hour fermentation period and a 24hr-sprouting period for millet arid pigeon pea may

be recommended for production of flour with high nutritive content.

Formulation of weaning formula from cereals like millet and legume like pigeon pea is

recommended to improve the protein quality of the cereals.

The diet IV fiom millet and pigeon pea blended in the ratio of 70:30 as shown in the

result of the animal bioassay is therefore recommended for use as weaning food for infants

particularly for the low income earners who can not afford commercial weaning foods.

5.3 Suggestion for Further Studies Like all intellectual endeavours, this research work generates additional studies. Hence,

we suggest that hrther studies be conducted to ascertain the nutritional quality of millet and

pigeon pea that have undergone the dual process of germination and fermentation.

To evaluate the essential amino acids in a cereal and the particular legumc that could

complement it through an essential amino acid assay.

To assay the effect of the millet and pigeon pea mixed protein diets on the animal organs

through a histopathological analysis.

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APPENDIX A

A

0 24 48 72 96

Fermentation time (hour)

C 'G 15 4-8

2 a 10 a, u 3

0 ' 5

- . - ~ ,. . . .. . . . - - . - . . . - .. r~ .~ ..-T --- ... ~ . . . - 1

0 24 48 72 96

Fermentation time (hour)

-- - 1: Crude protein (millet)

a C~ude protein (P. pea)

/ t crude fat (Millet) I 1 C~ade fat ( P pea) 1

It Crude fibre nille let) 1 1 Crude fibre (P. pea) 1

Fermentation time (hour)

Figure 1: Effect of Fermentation time on (A) crude protein, (B) crude fat and

(C) crude fibre content of millet and Digeon Dea.

(millet)

~ r u d e fat (millet)

Sprouting time (hour)

I 1

24 48 72 96

Sprouting time (hour)

3.5

3

- - - - --. - -. --- -- 2

L1 &Crude fibre (millet)

IE a 1.5 .-a- Crude fibre (P. pea)

Figure 2: Effect of Sprouting on (A) crude protein, (B) crude fat and (C)

TJ 3

G 1

0.5

0

crude fibre content of millet and pigeon pea. ,

I

0

Fermentation time (hour)

1 -+Tannin (millet) I

~ I -+ Phytate (millet) I

0 24 48 72 96

Fermentation time (hour)

a- Phytate (P. pea) 1 11 _

1% cyanide (millet) 1 1 --t Cyanide (P. .. . pea) - .. 1

Fermentation time (hour)

Figure 3: Effect of Fermentation time on (A) Tannin, (B) Phytate qnd (C) Cyanide content of millet and pigeon pea. , r l

Sprouting time (hour)

0.14

h

0.12

P 0.1 E' ... - 0.08 I-+ Phytate (millet)

g 0.06 1-6 Phytate (P. pea) 5;.

0." I

0.02

0 0 24 48 72 96

Sprouting time (hour)

Sprouting time (hour)

Figure 4: Effect of Sprouting time on (A) Tannin, (B) Phytate and (C) Cyanide content of millet and pigeon pea.

+Viscosity (Millet) --m- Viscosity (P. pea) .- -- - -. -

Fermentation time (hour)

+ WAI (Millet) *- WAI (P. pea)

. . 1 0 1 I I I I I

0 24 48 72 96

Fermentation time (hour)

+ WSI (Millet) + WSI (P.pea) 1

Fermentation time (hour)

Figure 5: Effect of Fermentation time on (A) Viscosity, (B) Water Absorption

Index and (C) Water Solubility Index of millet and pigeon pea.

Sprouting time (hour)

B

--

I-+-\liscosity (Millet) / /t ~ l s c o s i t ~ (P. pea)i

Sprouting time (hour)

I I I I I

0 24 48 72 96

Sprouting time (hour)

Figure6: Effect of Sprouting time on (A) Viscosity, (B) Water Absorption Index and (C) Water Solubility Index of millet and pigeon pea.

APPENDIX B CALCULATION OF THE COMPOSITION OF DIETS FED TO RATS:

No. of Rats -- 6 No. of Days - 28 Quantity of diets per day - 15 g/day Total weight of diet: 6 x 28 x 15 = 2520 Plain of Nutrition (Protein level) = 10% Based on 10% Plain of Nutrition, 7% of the protein in the diet will be supplied from pigeon pea) legume and 3% protein from millet (cereal).

Protein Content: Crude protein content of UPF = 22.04%

@.in5 To calculate the quantity of UPF that will give 7% protein, this expression used:

22.04 = 7 - 100 X

22.04~ = 700 x = 700 -

22.04 = 31.76

If 3 1.76 is equivalent to 7% protein level, To calculate the weight of UPF from the Total weight:

31.76 x 2520 100

= 800.35 g/g sample

Crude protein content of FPF,, = 23.46% acir To calculate the quantity of FPF7, that will give 7% protein, this expression b, used:

23.46 = - 7 100 X

23 .46~ = 700 X = 700

23.46 x = 29.84

If 29.84 is equivalent to 7% protein level, To calculate the weight of FPF,, from the Total weight:

29.84 x 2520 100

= 75 1.97 g/g sample

Crude protein content of SPFz4 = 24. 95% c * ( ( r i To calculate the quantity of SPF24 that will give 7% protein, this expression ,, used:

2 4 . 9 5 = 7 100 X

24 .95~ = 700 X = 700

24.95 x = 28.06

If 28.06 is equivalent to 7% protein level, To calculate the weight of SPF,, from the Total weight

28.06 x 2520 100

= 707.1 l g/g sample

Crude protein content of UMF = 9.18% To calculate the quantity of UMF that will give 3% protein, this expression is used:

9 . 1 8 = 3 - 100 X

9 . 1 8 ~ = 300 X = 300

9.18 x = 32.68

If 32.68 is equivalent to 3% protein level, To calculate the weight of UMF from the Total weight:

. 32.68 x 2520 100

= 823.54 g/g sample

Crude protein content of FMF,, = 9.46% i+i~ To calculate the quantity of FMFZ4 that will give 3% protein, this expression ,t, used:

9 . 4 6 = 3 - 100 X

9 . 4 6 ~ = 300 X = 300

9.46 x = 31.71

If 3 1.71 is equivalent to 3% protein level, To calculate the weight of FMFZ4 from the Total weight:

. 31.71 x 2520 100

= 799.09 g/g sample

Crude protein content of SM17,, = 7.88% a n 5 To calculate the quantity of SMFZ4 that will give 3% protein, this expression j\ used:

7 . 8 8 = 3 - 100 X

7 . 8 8 ~ = 300 x = - 300

7.88 x = 38.07

If 38.07 is equivalent to 3% protein level, To calculate the weight of SMF,, fiom the Total weight:

.: 38.07 x 2520 100

= 959.36dg sample

Crude protein content of Casein = 83.2% '4Q To calculate the quantity of casein that will give 7% protein, this expression used:

83.2 = - 10 100 X

8 3 . 2 ~ = 1000 X -

83.2 X

- - 12.019

If 12.0 19 is equivalent to 10% protein level, To calculate the weight of casein from the Total weight:

.: 12.019 x 2520 100 - 302.88 g/g sample

Fat content = 5% To calculate the 5% weight of fat from the Total weight

.: 0.05 x 2520 = 126

Mineralcontent = 3.5% To calculate the 3.5% weight of mineral fiom the Total weight

.: 0.035 x 2520 = 88.2

Vitamin content = I % To calculate the 1% weight of vitamin from the Total weight

.: 0.01 x 2520 = 25.2

Fibre content = 1% 'To calculate the 1% weight of fibre fiom the Total weight

.: 0.01 x 2520 = 25.2

Energy = 100 - (Protein + fat + mineral + vitamin + fibre contents ) = 100- ( lO+5+3 .5+1 + I ) = 100- 20.5 = 79.5

Diet I = 2520 - 1888.49 = 631.51

Sucrose and cornstarch supply energy, . The value for Energy is divided by 2

To give values for cornstarch and sucrose .: 631.5112 = 315.755

Diet I1 = 2520 - 181 5.66 = 704.34

.: Energy value for sucrose and cornstarch .: 704.34 I 2 = 353.17

Diet 111 = 2520 - 1975.93 = 544.07

.: Energy value for sucrose and cornstarch .: 544.07 1 2 = 272.035

Diet lV = 2520 - 1770.8 = 749.2

.: Energy value for sucrose and cornstarch . 749.2 12 = 374.6

Diet V = 2520 - 193 1.07 = 588.93

.: Energy value for sucrose and cornstarch . 599.93 12 = 294.465

Diet VII = 2520 - 567.48 - - 1952.52

.: Energy value for sucrose and cornstarch 1952.5212 = 976.26

APPENDIX C

DEPARTMENT OF FOOD SCIENCE & TECHNOLOGY UNIVERSITY OF NIGERIA, NSUKKA

SENSORY EVALUATION SCORE SHEET Name: Sex: Dept: Age: 30-30

30 4 0 40 -50

Please taste the products presented and score them according to your degree of acceptance using the scoring scheme provided.

Tick (4 ) to indicate acceptance 1 - 7 indicates the degree of acceptance; 7 = the highest, 1 = the least A - K shows the number of sam~les

Neither attractive nor unattractive 4 Slightly unattractive 3 Moderately unattractive 2 Extremely unattractive 1

I Any other comment:

1 ii. FLAVOUR I A I B I C I D I E I F I G I H 1 1 I J I K I Extremely strong 7 Moderately strong 6 Slightly strong 5 Neither strong nor weak 4 Slightly weak 3 Moderatelv weak 2

I I 1 1 I I I I I I

Extremely weak 1 I Anv other comment: I

I Anv other comment: 1

iii. FLAVOUR Extremely thick 7 Moderately thick 6 Slightly thick 5 Neither thick nor thin 4 Slightly thin 3 Moderately thin 2 Extremely thin 1

A B c D E F G J K -

other comment:

-

LAny other comment:

B C D E F G H I J K 1V. MOUTH FEEL y<tremely smooth 7 Moderately smooth 6 Slightly smooth 5

Neither smooth nor gritty 4 Slightly gritty 3 Moderately gritty 2 Extremely gritty 1

A

B C D E F G H I J K V. TASTE Extremely tasty 7 Moderately tasty 6 Slightly tasty 5 Neither tasty nor tasteless 4 Slightly tasteless 3

-Moderately tasteless 2 Extremely tasteless 1

L A ~ Y other comment: . .- --..--A

A

VI. AFTER TASTE Extremely strong 7 Moderately strong 6 Slightly strong 5 Neither strong nor weak 4 Slightly weak 3 Moderately weak 2 Extremely weak 1

A

VI. OVERALL ACCEPTABILITY Extremelv acce~table t 7

l ~ n y other comment:

B C D E F G H I J K

A

Moderately acceptable 6 Siightly acceptable 5 Neither acceptable nor unacceptable 4 Slightly unacceptable 3 Moderately unacceptable 2 Extremely unacceptable /:I 1

-

I I

-

B C D E F G H I J K ~ 1

I