EFFECT OF SUN DRYING ON THE NUTRITIVE

VALUE OF OKRA (Hibiscus esculents)

By

Almubark Musa Tibin Musa

B.SC. Agric. (Honours) (2005)

Faculty of Agriculture
Umdurman Islamic University

Dissertation submitted to the University of Khartoum in partial

fulfillment of the requirements for the Degree of Master of Science in
Food Science and Technology

Supervisor
Prof. El Gasim Ali El Gasim

Department of Food Science and Technology

Faculty of Agriculture
University of Khartoum

April 2008
 To my father and mother
For their continuous
Encouragement and blessings
And to whom I am always indebted
To my brothers, sisters and friends
For their tender love, care and encouragement
Hoping that we shall never separate
To all whom I love

i
 Acknowledgments
I am greatly indebted to my supervisor professor El- Gasim Ali El-
Gasim, for his keen guidance, assistance and continuous encouragement
during this work and for reading the draft manuscript.
Thanks and gratitude to the staff of Central Lab, Ministry of Science and
technology for material. The help of Prof. Abdelraheim Ahmed El
Hessian Dr Dina Ahmed For her helping, Azza babiker in statistical
analysis. My thanks are extended to staff of Biochemistry Department,
Central lab, Ministry of Science and technology Especial regards and
indebt ness are cordially extended to all Department members of Food
Science and Technology Department, Faculty of Agriculture, University
of Khartoum, for their great help and cooperation, and special regards to
my friend Abdelmonem Abdellah and Mahamoud Mohammed Siraj for
helps.
Best regard to my Father for helping me
Last but not least, my sincere thank and recognition to my family for
their helps.

ii
 Abstract
Two lines of Okra (Hibiscus esculentus) were collected to study the effect
of sun drying on the nutritive value of Okra.Chemical composition, anti
nutritional factors content, in vitro protein digestibility, amino acids and
minerals of Fresh and dried okra (hibiscus esculentus) was studied. Result
obtained showed that ash, crude protein, crude fiber and oil contents were
increased and the moisture was decrease after drying of okra pods.
Results obtained for anti nutritional factors showed that phytic acid
content of Fresh okra was found to be 33.96 mg/100g and after drying
decreased to 33.09 mg/100g, Tannin content of the fresh samples was
found to be 74.2 mg/100g and after drying it decreased to 22.55
mg/100g. Polyphenol content of the fresh samples was found to be 706.2
mg/100g and after drying decreased to 553 mg/100g. The protein
digestibility of fresh okra was found to be 30.72 % and after drying it was
increased to 82.57%. The result of amino acid analysis indicated that okra
is a good protein source of essential amino acid Drying was increases this
essential amino acid. essential amino acid content of fresh okra was
found to be Leucine , Valine , Arginine , Lysine , Isoleucine ,
Phenylalanine , Threonine , Histidine , Methonine 133.93,121.53,
119.77, 113.06, 90.46, 81.12, 77.57, 76.80, 35.47 mg/100g .respectively .
And after drying it was be increased and found to be 574.14, 547.08,
704.26, 346.98, 388.56, 326.28, 295.53, 322.15, 155.15mg/100g.
minerals content of fresh okra as found to be 0.069% , 0.26%, 0.36% ,
0.57% for Sodium , Phosphorus , Calcium , and Potassium
respectively,and after drying it increased to 0.09 %, 0.32%, 0.38%,
0.66% respectively.

iii
 ‫اﻟﺨﻼﺻﺔ‬
‫ﺗﻢ ﺟﻤﻊ ﺻﻨﻔﻴﻦ ﻣﻦ اﻟﺒﺎﻣﻴﺔ ﻟﺪراﺳﺔ ﻟﺪراﺳﺔ ﺗﺎﺛﻴﺮ اﻟﺘﺠﻔﻴﻒ اﻟﺸﻤﺴﻲ ﻋﻠﻲ اﻟﻘﻴﻤﺔ اﻟﻐﺬاﺋﻴﺔ‬
‫ﻟﻠﺒﺎﻣﻴﺔ ﺗﻤﺖ دراﺳﺔ اﻟﺘﺮآﻴﺐ اﻟﻜﻴﻤﻴﺎﺋﻲ وﻣﻀﺎدات اﻟﺘﻐﺬﻳﺔ وﻣﻌﺎﻣﻞ هﻀﻢ اﻟﺒﺮوﺗﻴﻦ واﻻﺣﻤﺎض‬
‫اﻻﻣﻴﻨﻴﺔ واﻟﻤﻌﺎدن اﻟﻤﺘﺎﺣﺔ ﻟﻠﻌﻴﻨﺎت اﻟﺨﺎم و اﻟﻤﻌﺎﻣﻠﺔ ﺑﺎﻟﺘﺠﻔﻴﻒ اﻟﺸﻤﺴﻰ ﻟﺼﻨﻒ اﻟﺒﺎﻣﻴﺔ ﺧﺮﻃﻮﻣﻴﺔ‬
‫)اﻟﺒﺎﻣﻴﺔ اﻟﻄﺎزﺟﺔ واﻟﻤﻌﺎﻣﻠﺔ ﺑﺎﻟﺘﺠﻔﻴﻒ اﻟﺸﻤﺴﻰ ( ‪.‬وأﻇﻬﺮت اﻟﺪراﺳﺔ أن ﻧﺴﺒﺔ اﻟﺮﻃﻮﺑﺔ وﻣﺤﺘﻮﻳﺎت‬
‫اﻟﺮﻣﺎد واﻟﺒﺮوﺗﻴﻦ اﻟﺨﺎم واﻷﻟﻴﺎف اﻟﺨﺎم واﻟﺪهﻦ زادت ﺑﻌﺪ اﻟﺘﺠﻔﻴﻒ‪ ،‬أﻣﺎ ﻣﻀﺎدات اﻟﺘﻐﺬﻳﺔ واﻟﺘﻲ‬
‫ﺗﺸﻤﻞ ﺣﻤﺾ اﻟﻔﺎﻳﺘﻚ ﻟﻠﺒﺎﻣﻴﺔ اﻟﺨﺎم واﻟﺬي آﺎن ‪33.96‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ وﺑﻌﺪ اﻟﺘﺠﻔﻴﻒ اﻧﺨﻔﻀﺖ إﻟﻲ‬
‫‪33.09‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ واﻟﺘﺎﻧﻴﻨﺎت ﻟﻠﺨﺎم ‪74.2‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ وﺑﻌﺪ اﻟﺘﺠﻔﻴﻒ اﻧﺨﻔﻀﺖ إﻟﻲ‬
‫ﻟﻠﺨﺎم آﺎﻧﺖ ‪706.2‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ وﺑﻌﺪ اﻟﺘﺠﻔﻴﻒ‬ ‫‪22.55‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ‪ .‬واﻟﻌﺪﻳﺪ اﻟﻔﻴﻨﻮﻻت‬
‫اﻧﺨﻔﻀﺖ إﻟﻲ ‪553‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ‪ .‬ﻣﻌﺎﻣﻞ هﻀﻢ اﻟﺒﺮوﺗﻴﻦ ﻟﻠﺒﺎﻣﻴﺔ اﻟﺨﺎم آﺎﻧﺖ ‪ %30.2‬وﺑﻌﺪ‬
‫اﻟﺘﺠﻔﻴﻒ زاد إﻟﻲ ‪ .%82.57‬اﻷﺣﻤﺎض اﻷﻣﻴﻨﻴﺔ ﻳﺘﻀﺢ ﻓﻲ آﻼ اﻟﺨﻄﻴﻦ أن اﻟﺒﺎﻣﻴﺔ ﻣﺼﺪر ﺟﻴﺪ‬
‫ﻟﻠﺒﺮوﺗﻴﻨﺎت وذﻟﻚ ﺑﺎﺣﺘﻮاﺋﻬﺎ ﻋﻠﻲ اﻷﺣﻤﺎض اﻷﻣﻴﻨﻴﺔ اﻷﺳﺎﺳﻴﺔ وﻗﺪ أدي اﻟﺘﺠﻔﻴﻒ إﻟﻲ زﻳﺎدة هﺬﻩ‬
‫اﻟﺤﻤﻮض اﻷﻣﻴﻨﻴﺔ‪ ،‬ﻓﻨﺠﺪ أن اﻷﺣﻤﺎض اﻷﻣﻴﻨﻴﺔ اﻷﺳﺎﺳﻴﺔ ﻓﻲ اﻟﻌﻴﻨﺔ اﻟﺨﺎم ﻟﻜﻞ ﻣﻦ ﻣﻦ اﻟﻠﻴﻮﺳﻴﻦ‪،‬‬
‫اﻟﻔﺎﻟﻴﻦ‪ ،‬اﻷرﺟﻨﻴﻦ‪ ،‬اﻟﻼﻳﺴﻴﻦ‪ ،‬اﻷﻳﺰوﻟﻴﻮﺳﻴﻦ‪ ،‬اﻟﻔﺎﻧﺎﻳﻞ‪ ،‬اﻟﺜﻴﺮوﻧﻴﻦ‪ ،‬اﻟﻬﺴﺘﻴﺪﻳﻦ‪ ،‬اﻟﻤﺜﻴﻮﻧﻴﻦ آﺎﻧﺖ‬
‫‪ 35.47 ،76.80 ،77.57 ،81.12 ،90.46 ،113.06 ،119.77 ،121.53 ،133.93‬ﻋﻠﻰ‬
‫اﻟﺘﻮاﻟﻰ ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪ .‬وﺑﻌﺪ اﻟﺘﺠﻔﻴﻒ زادت إﻟﻲ ‪، 346.98 ، 704.62 ، 547.08 ، 574.14‬‬
‫‪ 155.15، 322.15 ، 295.53، 326.28 ، 388.56‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻣﻦ اﻷﺣﻤﺎض اﻷﻣﻴﻨﻴﺔ‪.‬‬
‫اﻟﻌﻨﺎﺻﺮ اﻟﻤﻌﺪﻧﻴﺔ اﻟﻐﺬاﺋﻴﺔ ﻟﻠﺒﺎﻣﻴﺔ اﻟﻄﺎزﺟﺔ ﻟﻜﻞ ﻣﻦ اﻟﺼﻮدﻳﻮم واﻟﻔﺴﻔﻮر واﻟﻜﺎﻟﺴﻴﻮم واﻟﺒﻮﺗﺎﺳﻴﻮم‬
‫آﺎﻧﺖ ‪ %0.57 ، %0.36، %0.26 ، %0.069‬ﻋﻠﻰ اﻟﺘﻮاﻟﻰ ‪ .‬وﺑﻌﺪ اﻟﺘﺠﻔﻴﻒ زادت إﻟﻲ ‪0.09‬‬
‫‪ 0.66، 0.38، 0.32 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻣﻦ اﻟﻌﻨﺎﺻﺮ اﻟﻤﻌﺪﻧﻴﺔ اﻟﻐﺬاﺋﻴﺔ ﻋﻠﻲ اﻟﺘﻮاﻟﻲ‪.‬‬

‫‪iv‬‬
 TABLE OF CONTENTS

CONTENTS Page
Dedication i

Acknowledgements ii

Abstract iii

Arabic Abstract vi

List of Contents vii

List of Table viii

CHAPTER ONE: INTRODUCTION 1

CHAPTER TWO: LITERATURE REVIEW 4

2.1 Chemical composition 4

2.1.1 Moisture content 4

2.1.2 Crude protein content 5

2.1.3 Fat content 6

2.1.4 Ash content 6

2.1.5 Fiber Content 7

2.2 Minerals 7

2.2.1 The bioavailability of minerals elements 8

2.2.2 Potassium and sodium content of okra 9

2.2.2.1 Potassium 9

2.2.2.2 Sodium 9

2.2.3 Calcium and Phosphorous contents of Okra 10

2.2.3.1 Calcium 10

2.2.3.2 Phosphorous 11

2.3 Amino Acids 11

2.3.1 The Essential Amino acids 12

2.3.1.1 Isoleucine 12

2.3.1.2 Leucine 12

v
2.3.1.3 Valine 13

2.3.1.4 Histadine 13

2.3.1.5 Lysine 13

2.3.1.6 Methionine 13

2.3.1.7 Phenylalanine 13

2.3.1.8 Threonine 14

2.3.1.9 Tryptophan 14

2.3.2 Conditionally Dispensable Amino Acids 14

2.3.2.1 Arginine 14

2.3.2.2 Cysteine 14

2.3.2.3 Tyrosine 14

2.3.3 Nonessential Amino Acids 15

2.3.3.1 Alanine 15

2.3.3.2 Aspartic Acid 15

2.3.3.3 Cystine 15

2.3.3.4 Glutamic Acid 15

2.3.3.5 Glutamine 16

2.3.3.6 Glycine 16

2.3.3.7 Ornithine 16

2.3.3.8 Proline 16

2.3.3.9 Serine 16

2.3.3.10 Taurine 17

2.4 In vitro Protein Digestibility (IVPD) 17

2.5 Anti-nutrition factors of fresh and dried okra 17

2.5.1 Tannins content 17

2.5.2 Total polyphenols content 18

2.5.3 phytic acid 18

vi
CHABTER THREE :MATERIALS AND METHODS 20

3.1 Chemical composition 20

3.1.1Moisture content 20

3.1.2 Crude protein 21

3.1.3 Fat content 22
3.1.4 Ash content 23
3.1.5 Crude Fiber 24
3.2 Determination of mineral 25
3.2.1 Determination of Total minerals 25
3.2.2Phosphorus determination 25
3.3.2 Amino acid Determination 27
3.4 In vitro protein digestibility determination 29
3.5 Anti-nutritional factors determination 30
3.5.1 Determination of phytic acid 30
3.5.2 Determination of tannin content 31
3.5.2.1 Tannin standard curve 32
3.5.3 Total polyphenol (TP) determination 33
3.5.3.1 Standard curve preparation 33
CHAPTER FOUR :RESULTS AND DISCUSSION 35
4.1 Chemical Composition 35
4.1.1 Moisture Content. 35
4.1.2 Protein content 35
4.1.3 Fat content 35
4.1.4 Ash content 37
4.1.5 Fiber content 37
4.2 Minerals composition 37
4.3 Amino Acids profile 39
4.4 Protein Digestibility 41

vii
4.5 Anti-nutritional Factors: 41
CHAPTER FIVE :CONCOLUSIONS AND RECOMMENDATIONS 44
Conclusions 44
Recommendations 44
REFERNCES 45
 

viii
  

LIST OF TABLES

TABLE Pages
1. Chemical Composition of Okra 36

2. Minerals Content of Fresh and Dried Okra 38

3. Amino Acid Profile of Fresh and Dried Okra 40

4. In vitro Protein Digestibility of Fresh and Dried Okra. 42

5. Anti Nutritional Factors in Okra 43

iv
 CHAPTER ONE

INTRODUCTION

Vegetables play a very important role in the human diet

(Thompson and Kelly, 1957). They have been valued for their health–
giving qualities. Certain vegetables are rich sources of minerals needed
for body building and body regulating: some are good source of fat,
proteins and carbohydrate; others are rich source of vitamins.

Okra, (Hibiscus esculentus), is a vegetable that has been used

extensively in the Sudan as a major traditional diet, yet little basic
chemical research has been done on it. It originates probably from East
Africa and to day is widely distributed in the tropics, subtropics and
warmer portion of the temperate region (ECHO, 2003).

The economic importance of okra can not be over emphasized;

It contains carbohydrate, proteins, and vitamin C in large quantities
(Adeboye and Aoputa, 1996). The essential amino acids that okra
contains are comparable to that of soybean. Hence, it plays a vital role in
human diet. Young immature fruits are of high nutritional value and can
be consumed in different forms .They could be boiled, dried, fried or
cooked. Okra is usually boiled in water resulting in slimy soups and
sauces, which are relished. The fruit also serve as soup thickeners
(Schippers, 2000). The leaves buds and flowers are also edible. Okra
seeds could be dried and the dried seeds can be either used to prepare
vegetable cruds, or roasted and ground to be used as coffee additives or
substitute.

Okra leaves are considered good cattle feed, but this is seldom
compatible with the primary use of the plant. Okra mucilage is suitable
1
for medicinal and industrial application. It has medically found
application as plasma replacement or blood volume expander.
Industrially, okra mucilage is usually used in to glace certain papers and
also useful in confectionery among other uses (Markose and Apeter,
1990).

Worldwide production of okra as fruit vegetable is estimated as six

million tones per year. In western Africa, it estimated at 500.000 to
600.000 tones per year (Burkil, 1997). In Nigeria there are two distinct
seasons for okra, the peak and the lean season. During the lean seasons
okra fruit are produced in low quantity, scarce and expensive to get
(Bamireand Oke, 2003). In the peak season, it is produced in large
quantities much more than what the local population can consume. Proper
processing, preservation, marketing and utilization of okra are necessary
to arrest the wastage being experienced during the peak season. Such
efforts should involve the development of appropriate technologies for
processing and preservation okra to produce products of high market
value. This will enhance the living standard of the farmer's and
processors. It is however, important to assess the level of okra to
production and preservation to serve as a guide in selecting appropriate
methods and techniques.

In Sudan Okra is the major popular vegetable. It is cultivated

widely for its fruits. The leaves of the plant can also be cooked as pot
herp either in the green or dried state. (Cobley, 1967).

Okra, fresh or dried, in combination with sorghum bread (kisra) is

the staple food of the Sudanese people. The wide type (Sara) is usually
used as a dry powder while the cultivated type is mainly used fresh.
Efforts have been directed to utilize the great natural variability found in
2
the rain –fed-and irrigated types of okra in the country. Some lines from
these types showed outstanding performance compared with introduced
varieties (Geneif, 1986). Three lines were selected for their high yield and
superior market qualities. These were released by the ARC (Agrecultral
Research Center ) in 1987 under the names Raiba, Higairat and Sennar
(Mohammed, 1991).

Objective:

1- To determine the nutritional quality of fresh and dried okra.

2- To determine the Anti-nutritional Factor in fresh and dried Okra.
3- To determine the effect of drying on chemical composition of Okra.

3
 CHAPTER TWO

LITERATURE REVIEW

2.1 Chemical composition

Chemical composition of plant material consists of determining the

major classes of chemical components which include moisture, crude
fiber, fat content (ether extracts), crude protein, ash and carbohydrates
(by difference).

Proximate composition provides a good initial impression of relative

nutritive value and utility of an agricultural products and allows basis of
comparison between different species, plant part and cultivation condition

2.1.1 Moisture Content

The determination of moisture content of food stuff is very important

for both commercial and scientific applications. The moisture content in
food is so difficult to be determined accurately. (Standard Committee,
1979). EL Awad (1980) found that the moisture contents of vegetable
ranged from 66.8 to 96.6, %. Ahmed (1986) determined the moisture
content of vegetables in wider range from 77.2 to 95.6 g / 100 g. Ahmed
(1998) investigated some vegetables and reported that moisture content
range from 65.3 to 95.4 %. Yousif (1993) mentioned that moisture
content of okra was 76.95%. Boutros (1977) reported that the moisture
content of okra pods 90.5 g./100g AL-Wandawi (1983) reported that the
moisture content of okra seeds 6.96 % for Emerald variety and 17.66 %
for Ibtaira variety. Also Watt and Merril (1963) found that the Moisture
content of okra seeds 7.92 %. FAO (1970) reported that the moisture
content of okra fruit is 88.6 g /100 g.
4
2.1.2 Crude protein

Proteins play a central role in biological systems. In addition to

functioning as enzymes, proteins also function as structural components
of cell in complex organisms. They are highly complex polymers,
colloidal in nature, and made up of 20 amino acids. Different proteins
vary in molecular weight and amino acids sequence. At the elemental
level proteins contain 50-55% C, 6-7% H, 20-23% O, 12-19% Na and
0.2-3.0% S (Damodaran, 1996). Proteins are either homoproteins or
heteroproteins conjugated with nonprotein component. Structurally,
protein also can be classified into globular and fibrous.

The role of protein in the diet cannot be overemphasized, since

protein is the basis of life. Protein deficiency has been recognized for
more than a century as a major nutritional problem in the world. Since
1936, a number of international groups, including the League of Nations,
FAO, WHO, the United Nations University (UNU) and the international
Dietary Energy Consultative group (IDECG), have made
recommendations on the protein requirements of infants, and children.

Protein requirement is defined by the FAO/WHO/UNU (1985) group

as the lowest level of dietary protein intake that will balance the losses of
nitrogen from the body in persons maintaining energy balance at modest
levels of physical activity. In children and pregnant or lactating women,
the protein requirements for children have changed significantly over the
years. In1953, 3-4 g/Kg per day of protein were recommended for
children between about six months and three years of age NAS-
NRC(1953), whereas now the recommended level is about 1g/kg per day
(IDECG 1996).these changes have significantly affected views on the
causes of child malnutrition in developing countries. So protein quality is
5
important. FAO (1970) reported that protein content of okra fruit 14.2 %.
Boutros (1977) reported the crude protein 2.3 g / 100 g. Edward and
Miller (1947) analyzed samples of okra seed meal and found that protein
content 13.56 % Watt and Merril (1963) reported that whole dry okra
seed contained 20.58 % protein. AL-Wandawi (1983) reported that crude
protein of okra seed 21.82 % for Emerald variety and 17.66 for Ibtaria
variety.

2.1.3 Fat Content

Fat are heterogeneous compounds, which are classified according

to their solubility in organic solvents as chloroform, ethyl ether,
petroleum ether or benzene. This solubility differentiates them from other
constituents such as proteins, carbohydrates and nucleic acids in seeds.
Lipids include free fatty acids, mono-glycerides, di-glycerides, tri-
glycerides, phospholipids, sterol ester, glycerols or glycerides. Boutros
(1977) reported that the fat content of okra 0.1 g / 100 g. Edward and
Miller (1947) analyzed samples of okra seed meal and found that the fat
content to be 1.92%. Watt and Merril (1936) reported that fat content of
okra seed 20.06 %.

2.1.4 Ash Content

The ash content of food stuff is the inorganic residue that remains
after burning the organic matter, is used as starting point for
determination of elemental composition of food material. However,
mineral constituents of ash are recognized to perform essential function in
human nutrition. They required by human body for good healthy and
growth. EL Awad (1980) found that the ash contents of vegetable ranged
from 0.4 to 2.3 %, Ahmed (1998) investigated some vegetable and he

6
reported that ash content ranged from 0.2 to 1.9 %. Yousif (1993)
reported that ash content of okra 8.3 %. Boutros (1977) reported that ash
content of okra 1.3 g / 100 g. Watt and Merril (1963) reported that the ash
content of okra seeds 8.19 %. And AL-Wandawi (1983) mentioned that
ash content of okra seeds 4.33 % for Emerald variety and 17.66 for
Ibtaira variety.

2.1.5 Fiber Content

Scientists defined dietary fiber as the protein of food which is derived

from cellular walls of plants which is digested very poorly by human
beings (Gordon, 1991). It consists of two major fractions, insoluble and
soluble dietary fiber. Soluble dietary fiber (SDF) is of particular interest
to many consumers because of its effects on blood cholesterol (Jenkins et
al., 1993; and Marllett and Cheung, 1997), Blood glucose (Wood et al.,
1994; and Marllett and Cheung, 1997), and prevents heart diseases
(Gordoen, 1991). Insoluble dietary fiber has been linked to protection
against colon cancer and other bowel disorders, such as constipation
(Marrlett and Cheung, 1997).On the other hand, crude fiber is found to
influence the rate of digestion by decreasing the activity of proteolytic
enzymes (Schneeman, 1990) the fiber content of Hibiscus esculentus is
1.6 g / 100 g reported by Boutros (1977) and Edward and Miller (1947)
analyzed samples of okra seed meal and found that the crude fiber (8.14
%) AL-Wandawi (1983), reported that the crude fiber of okra seed 27.30
% for Emerald and 27.20 for Ibtaira variety.

2.2 Minerals contents

The minerals constituents matter comprises a large number of

inorganic elements present in food stuff with varying amount
7
(Karakoltsidis and Constantinides, 1975). Part of these are recognized to
perform essential function in human nutrition, therefore, they must be
continuously supplied by food-stuff. Accordingly, 14 different mineral
elements are essentially required by animal body for good health and
growth (Wilson, 1967), Ca, P, k, Mg, Na, S and Cl are needed in
appreciable amount, i.e., macro-element. Other such as Fe, Cu, Mn, I, Co
……etc. Are required in trace elements are considered to play tow main
roles in animal body: as building constituents and as regulating
substances. On the basis of quantitative considerations, essential minerals
can be classified into major and trace element .Okra seed seem to contain
four major element, namely, Na, K, Ca, and Mg, and five trace elements
namely, Fe, Cu, Mn , Zn, and Ni the values reported for the element is
found to be potassium . The seeds also contain abundant amount of the
elements Na, Mg, Ca, Fe, Zn, Mn, and Ni. The value reported for the
elements Ca and Fe are in good agreement with those

2.2.1 The bioavailability of minerals elements

Some anti-nutritional components such as phytate if present in

food stuff may impair the absorption of the essential element, by binding
with them, thus rendering these elements non-available (Erdman, 1979).
Moreover, an antagonistic action of certain element may hamper the
absorption of specified elements .In instance, a large intake of Fe and Mg
may interfere with phosphorous absorption through binding to form
insoluble phosphates.

8
2.2.2 Potassium and sodium content of okra

The human body contains approximately 1 % K and 0.4 % Na.

potassium exists primarily as cellular constituent. Contrarily, much
sodium is concentrated in extra cellular fluids.

2.2.2.1 Potassium

Food source of potassium include all vegetables especially green

leafy vegetables (Ahmed, 1998; Rajalakshmi and Soikantia, 1980). The
widest dietary source of potassium is unprocessed foods, especially fruit
and many vegetables (Williams, 1995). Generally, food of plant origin
are rich in potassium, and it ranged from 50 .0 to 1700.0 mg / 100 g (Paul
and Southgate, 1981). Yousif (1993) examined the potassium content of
okra. She found that contained 1642 mg /100g. Al-wandawi (1981)
reported the potassium content of defatted seed flour for Emerald variety
to be 1309 .00mg / 100g for Emerald variety and 1591.40 mg / 100 g for
Ibtaira variety.

2.2.2.2 Sodium

Most foods of vegetable origin have comparatively low in sodium

content (Hulume, 1979). According to Paul and Southgate (1981), sodium
content of vegetable ranges from 00.0 to 1190 mg /100 g. Yousif (1993)
found that the sodium content of okra to be 80.0 mg / 100g. Al-wandawi
(1981) reported the sodium content for Emerald variety to be 647.20 mg
/100g and 475 .60 mg / 100 g for Ibtaira variety.

9
2.2.3 Calcium and Phosphorous contents of Okra

Calcium and phosphorous, quantitatively, are believed to be the

major constituents of the elementary composition of the human body (4
% Ca and 2.5 % P). They are closely associated with each other in most
parts and therefore inadequate supply of either in the diet limits the
nutritive value of both. Accordingly, calcium and phosphorus levels in
the blood are important indicators of the state of nutrition. More over, it
should be emphasized that their levels represent a balance between
several opposing factors: absorption – as understood that half or more of
the phosphorous of most seeds and their product is so combined to
phytin-, excretion, deposition and mobilization (Maynard and Loosli,
1962).

2.2.3.1 Calcium

Calcium is present in relatively high concentration in most diary

and diary products, cereal, nuts, fresh eggs and certain vegetables
(Pomeranz and Meloan, 1992). El Awad (1980) found that the calcium
content of vegetables on fresh basis ranged from 9.0 to 429.0 mg / 100g.
Raghuvanshi et al., (2001) reported a range of 19 to 355 mg / 100g for
some vegetable and concluded that they consider as good source of
macronutrients. Yousif (1993) reported the calcium content of okra to be
483.3 mg /100g. Boutros (1977) found that the Calcium content of Okra
152 g/100g. Watt and Merril (1963). Reported that the calcium content of
okra seed 282.26 mg /100g. Al-wandawi (1983) reported that the Calcium
content for Emerald variety to be 375.5 mg / 100g and for Ibtaira variety
268.8 mg/100g.

10
2.2.3.2 Phosphorous

Phosphorus servs as a partner with calcium in the major task of bone

formation, besides the other metabolic processes (Williams, 1995). El
Awad (1980) showed that the phosphorus content of vegetable ranged
from 4.0 to 99.0 mg /100g on fresh basis. However, he found that
phosphorus content of dried Okra reached 144.0 mg /100g. Okra
determined on by Yousif (1993) and was found to be 21.33 mg / 100g.
Boutros (1977) found that the phosphorous content of Okra 99 g/100g.

2.3 Amino Acids

The 23 or so amino acids are the molecular building blocks of

proteins. According to one accepted classification, 9 are termed essential
amino acids, meaning that they must be supplied from some food or
supplement source; the others, which used to be classified simply as
nonessential, are now more correctly termed nonessential amino acids,
essential and non essential based on the body's ability to synthesize them
from other amino acids.

You may not give it much thought when you sink your teeth into a
chicken breast (or lentil stew), but the content and balance of amino
acids, particularly the ratio of essential to, non essential is what
determines the body and health building value of a protein food or
supplement. But that isn't all that matters.

In addition to being influenced by the carbohydrates, fats and total

calories associated with it, protein quality is related to the amount of the
specific amines within both the essential and non essential categories (for
example, the amount of glutamine and branched chain amino acids, or

11
BCAAs - leucine, isoleucine and valine). While the amounts of essential
amino acids are generally of greater importance, the nonessential are also
significant because they're synthesized too slowly to support maximum
growth. Even if a source has a perfect amino acid profile for a given
individual and lifestyle, another important factor - to what extent these
acids are actually delivered to the tissues when needed - must be
considered. That, in turn, raises the issues of digestion, absorption, actual
bioavailability and the potential value of supplementation. According to
Finnin and Peters (1996) there are three types of amino acids; the
essential amino acids, the conditionally dispensable amino acids, and the
non essential amino acids. Essential amino acids must be supplied to the
body from food or supplements. Conditionally dispensable amino acids
are based on the body's ability to actually synthesize them from other
amino acids. Nonessential amino acids can be synthesized by the body
from other amino acids. Here is the amino acid guide and their benefits.
Finnin and Peters (1996)

2.3.1 The Essential Amino acids

2.3.1.1 Isoleucine

A branched chain amino acid readily taken up and used for energy by
muscle tissue. In debilitated individuals, isoleucine is known to prevent
muscle wasting. Moreover it is essential in the formation of hemoglobin

2.3.1.2 Leucine

A branched chain amino acid used as a source of energy and helps

reduce muscle protein breakdown .Also; it promotes healing of skin and
broken bones.

12
2.3.1.3 Valine

A branched chain amino acid, which is not processed by the liver; but
rather actively taken up by muscle

2.3.1.4 Histadine

One of the major ultraviolet absorbing compounds in the skin, It is

important in the production of red and white blood cells; used in the
treatment of anemia. Also, it is used in the treatment of allergic diseases,
rheumatoid arthritis and digestive ulcers

2.3.1.5 Lysine

Low levels can slow protein synthesis, affecting muscle and

connective tissue. Additionally, lysine is known to Inhibits viruses; used
in the treatment of herpes simplex, and aids bone growth by helping form
collagen, the fibrous protein that makes up bone, cartilage and other
connective tissue.

2.3.1.6 Methionine

May increase antioxidant levels (glutathione) and reduce blood

cholesterol levels. Also it Helps remove toxic wastes from the liver and
assists in the regeneration of liver and kidney tissue.

2.3.1.7 Phenylalanine

The major precursor of tyrosine, It Enhances learning, memory, mood

and alertness, It is used in the treatment of some types of depression but it
Suppresses appetite too. From the other hand it was found to be a major
element in the production of collagen.
13
2.3.1.8 Threonine

Generally low in vegetarians, it helps prevent fatty buildup in the liver,

as well as it is important component of collagen.

2.3.1.9 Tryptophan

Precursor of key neurotransmitter serotonin, which exerts a calming

effect, it stimulates the release of growth hormones. It is only available in
natural food sources, and free form of this amino acid is unavailable in
the U.S.

2.3.2 Conditionally Dispensable Amino Acids

2.3.2.1 Arginine

Can increase secretion of insulin, glucagon, growth hormones It

aids in injury rehabilitationm formation of collagen and immune system
stimulation. Additionally, arginine may increase sperm count and T-
lymphocyte response.

2.3.2.2 Cysteine

Detoxifies harmful chemicals in combination with L-aspartic acid and

L-citruline. It helps prevent damage from alcohol and tobacco use and
stimulates white blood cell activity.

2.3.2.3 Tyrosine

Precurso of the neurotransmitters dopaminem, norepinephrine and

epinephrine, as well as thyroid and growth hormones and melanin (the
pigment responsible for skin and hair color).

14
2.3.3 Nonessential Amino Acids

2.3.3.1 Alanine

Major component of connective tissue, As well as helping build up the

immune system. It was found that alanine is a key intermediate in the
glucose alanine cycle, which allows muscles and other tissues to derive
energy from amino acids.

2.3.3.2 Aspartic Acid

Helps convert carbohydrates into muscle energy and reduces ammonia

levels after exercises. Also, aspartic acid, builds immune system
immunoglobulins and antibodies.

2.3.3.3 Cystine

Contributes to strong connective tissue and tissue antioxidant actions,

In healing processes, it stimulates white blood cell activity and helps
diminish pain from inflammation. Also it is essential for the formation of
skin and hair.

2.3.3.4 Glutamic Acid

amajor precursor of glutamine, proline, ornothine, arginine,

glutathione, and GABA. a potential source of energy and important in
brain metabolism and metabolism of other amino acid

15
2.3.3.5 Glutamine

Most abundant amino acid, It plays a key role in immune system

functions, besides; it is an important source of energy, especially for
kidneys and intestines during caloric restrictions.

2.3.3.6 Glycine

Aids in the manufacture of other amino acids and is a part of the structure
of hemoglobin and cytochromes enzymes involved in energy production.
Also, glycine has a calming effect and is sometimes used to treat manic
depressive and aggressive individual’s .Furthermore it produces
glucagon, which mobilizes glycogen, and can inhibit sugar cravings.

2.3.3.7 Ornithine

May help increase growth hormone secretion in high doses. Also

ornithine found to aid in immune, liver function, and promotes healing.

2.3.3.8 Proline

A major component in the formation of connective tissue and heart

muscle, It is readily mobilized for muscular energy and it is considered as
a major constituent of collagen.

2.3.3.9 Serine

Important in cells' energy production, It aids memory and nervous

system function. Moreover, it helps builds up immune system by
producing immuno-globulins and antibodies.

16
2.3.3.10 Taurine

Aids in the absorption and elimination of fats, as well as it may act as a

neurotransmitter in some areas of the brain and retina.

2.4 In vitro Protein Digestibility (IVPD)

The nutritive of protein, referred as proteins quality, depends on its

content of amino acids and the in vitro digestibility (Hahn et al., 1984).
The percentage of an essential amino acid that is in greatest deficiency as
compared to the amount in standard or reference protein i9s Known as the
limiting amino acid or chemical score of protein . However, today it is
more common for scores to be calculated as percentage of adequacy
rather than percentage deficiency.

The protein digestibility is determined to provide the most satisfactory

indication of seed utilization (FAO / WHO / UNV, 1985).

The protein quality of food or feed is defined by its amino acid

composition and its digestibility. The protein digestibility primarily
determines the availability of its amino acids (Hahn et al., 1981).

2.5 Anti-nutrition factors of fresh and dried okra

2.5.1 Tannins content

Tannins are believed generally, to play a significant role in nutrition by

binding the most essential nutrients, thus altering their chemical and
functional properties. Tannin has been extensively investigated because
of their harmful effects on growth of animals and Protein utilization
thought formation of insoluble complexes with them. This process is
responsible of reducing the digestibility of proteins, and consequently the

17
protein value of food (McLeod, 1974). As so much tannin have found to
exhibit the ability to inactivate several enzymes (Strumeyer and Malin,
1975)

2.5.2 Total polyphenols content

Phenolic compounds embrace a wide range of plant substances

which posses an aromatic ring bearing one or more hydroxyl constituents
(Harbone, 1962). Polyphenols constitute tannin a level varies even
between the cultivars of the same species. Genetic factors, as will as
environmental conditions largely influence their presence in plant foods.
Several functions have been attributed to polyphenols; they have anti-
pathogenic, anti-herbivore and allelopathic properties (Brice and
Morrison, 1982; Ray and Hastings, 1992). Phenolic compounds are
partially responsible for the sensory and ant nutritional quality of plant
foods (Bravo, 1998).

2.5.3 Phytic acid

Phytate represents a complex class of naturally occurring phosphorus

compounds that can significantly influence the functional and nutritional
properties of foods. Although the presence of these compounds has been
known for over a century, their biological role is not completely
understood. Phytic acid, myo-inositol 1, 2, 3, 4, 5, 6-hexaphosphoric acid
(dihydrogen phosphate), is the main phosphorus store in mature seeds.
Phytic acid has a strong binding capacity, readily forming complexes
with multivalent cations and proteins. Most of the phytate-metal
complexes are insoluble at physiological PH. Hence phytate binding
renders several minerals biologically unavailable to animals and humans.

18
 The effects of phytic acid in human and animal nutrition are related
to the interaction of phytic acid with proteins, vitamins and several
minerals, thereby restricts their bio-extractability. In view of the anti-
nutritional effects of phytic acid, many attempts to reduce phytate have
been made. phytic acid is said to chelate mineral cations and proteins,
forming insoluble precipitates, which leads to reduced bioavailability of
trace mineral cations and reduced digestibility of proteins (Ryden and
Selevndran, 1993).

19
 CHAPTER THREE

Materials and methods

3.1 Chemical composition

3.1.1 Moisture content

The moisture content was determined according to the method of

AOAC (1995). Two grams of well mixed ground sample were accurately
weighed in an uncovered clean, dry dish of known weight. It was then
heated in an oven provided with a fan at 125oC for 4 hours. The dish was
then covered and transferred to a desicater. After cooling at room
temperature, it was weighed again. The loss of weight was calculated as a
percentage from the sample weight and expressed as % moisture content.

Moisture content W1 – W2
X 100
(%) = W

-W1= Weight of sample + crucible before oven drying

-W2=Weight of sample + crucible after oven drying

- W= Weight of sample.

20
3.1.2 Crude protein

The nitrogen content was determined by the microkjeldhal method

as described by Pearson (1981).

Digestion procedure

About 0.5 g of sample was transferred into kjeldahl digestion flask.

The sample was neutralized with 10ml of concentrated sulphuric acid and
one tablet of the catalyst (potassium sulphate and mercuric oxides) was
added per digestion. The flask was heated moderately initially in a fume
hood and shaken from time to time until the mass was carbonized and the
frothy had disappeared. The heat applied was then increased and the
liquid was brought to a steady boiling. When the solution appeared clear
the boiling was continued for further one hour and then the solution was
allowed to cool and rapidly 5 ml of water were added. The digested
sample was transferred too 250 ml volumetric flask.

Distillation procedure:

Reagents:

- Bromocresol green –methyl red (0. 4 g) and methyl red (0.15 g)

were dissolved together as an indicator.

- Sodium hydroxide solution 20ml (33%).

- Hydrochloric acid solution 0.1 N aqueous solution

Procedure:

Aliquot of the digested sample was transferred to the Markham

distillation unit. 20 ml of 33% sodium hydroxide were added to make the
solution strongly alkaline, the funnel was then washed with few mls of
21
distilled water and the steam generator turned on. The ammonia was
collected in a 100 ml conical flask containing 50ml of 2% Boric acid and
four drops of mixed indicators .After three minutes the distillation was
stopped.

Titration procedure:

-The solution was titrated against 0.1N HCl

-The total crude protein was calculated from the equation below

T x 0.0014 x 100
C. N%=
W.S

T x 0.14 x 6.25
C. P%=
W.S

Where:

-T = Volume of sample titrate

-W.S = Weight of the sample

-6.25 = Protein factor to convert nitrogen to crude or generalized

protein.

-14 = Equivalent weight of nitrogen

1000 = Number of milligrams in one gram

3.1.3 Fat content

The total oil content of the sample was determined according to

AOAC (1995). Two grams of the ground sample were accurately
weighed in an empty thimble and was then plugged with a piece of
22
cotton, the solvent, petroleum ether B.P. (40 – 60oC) was poured into a
pre-weighed round bottomed 150 ml flask (up to 2/3 of the flask volume).
The soxhlet apparatus was then assembled and the sample was extracted
for 4 hours using a steam bath. After carefully dismantling the apparatus,
the solvent was evaporated to dryness using a rotatory evaporator. The oil
content was calculated according to the equation below:

Weight of the oil extracted in gm X 100

Oil content (%)
W.S

- W.S = Weight of the sample

3.1.4 Ash content

The method described by AOAC (1995) was used. Two grams

of the sample were accurately weighed into a weighed porcelain dish, and
dried at 100oC for 3 – 4 hours using drying oven. The dish and content
were then transferred to a muffle furnace and ignited at 500 – 600oC and
let to stay overnight or until free from carbon (residue appears grayish
white). The dish and content were then removed and placed into a
desicater. After cooling, it was weighed and the results were calculated
and expressed as below

Ash content (%) = (B – C)

X 100
A

Where:

-A = Sample weight (g)

-B = Weight of dish and ash content (g)

23
 -C = Weight of empty dish (g)

3.1.5 Crude Fiber

The crude fiber content was carried out by the method of AOAC (1984)
2.7 to3.0g of the dried and defatted sample, were transferred to a 600 ml
beaker with a few anti-bumping granules. The sample was digested with
200 ml of .255 N sulphuric acids for exactly 30 minutes, and the beaker
was periodically swirled. The contents were removed and filtered through
Buchner funnel, and washed with boiling water. The digestion was
repeated using 200ml of .313N sodium hydroxide for 30 minutes, and
treated similarly as above. After that the fiber was washed with 1%
hydrochloric acid to neutralize the sodium hydroxide and then rinsed with
distilled water. After the last washing the residue was transferred to
ashing dish, and dried in an oven at 103˚C for one hour then cooled and
weighed. The dried residue was ignited in muffle furnace at 500 ˚C over
night, cooled and reweigh. The crude fiber was calculated using

The following equation:

CF% = W1- W2 x10

WS

Where:

-CF% = crude fiber

-WS =weight of sample

-W1 =weight of crucible with sample.

24
-W2 = weight of crucible with ashed sample

3.2 Determination of mineral

3.2.1 Determination of total minerals

Total minerals which include Na, K, Ca, and P were determined

according to the method Champan and part (1982) as described below:

Two grams of sample were weighed in a clean dry crucible. The

crucible was placed in a muffle furnace for 4h at 550 C°, samples were
cooled and 10 ml of 5N HCl was added, covered with watch glass and
boiled gently for 10 minutes using sand bath, then cooled. The contents
were filtered through Whatman filter paper No. 4 and the volume was
made to 50 ml with distilled water, and were taken for mineral
determination either by titration for Ca and Mg determination or by
atomic absorption spectrophotometer (perkin- Elmer 2380)

Mg mineral \100g sample = mg\L X volume used X 100

1000 X wt. of sample

Where:

-Mg\L = ppm (curve reading)

-Volume used = Volume of extract

-1000 = Conversion from mg\L to mg\ml

-Wt. of sample = Two grams

3.2.2 Phosphorus determination

The phosphorus for both total and HCl- extractable for each sample
were carried out according to the method of Chapman and Pratt (1982).
25
One ml of the ash extract was pipetted into a 50 volumetric flask. Ten ml
of the ammonium molybdate ammonium vanadate reagent (22.5g of
(NH4)6 MO7 O24 4H2O in 400 ml distilled water+ 1.25g ammonium
vanadate in 300 ml boiling water + 250 ml con. HNO3 then diluted to one
Liter) were added.

The contents in the flask were mixed and diluted to volume. The density
of the color was read after 30 minutes at 470 nm using spectrophotometer
(JENWAY 6305 UV/Vis.).

Phosphorus was determined from standard curve.

Calculation:

P (mg/100g) = Mg/L X vol. Use X100

1000 X wt. of sample

Where:

-Mg/L = ppm (curve reading)

-Volume used = volume of extract

- 1000 = conversion from mg/L to mg/ml

-Wt of sample = 2grams

A standard and curve of phosphorus was plotted according to

Chapman and Pratt (1982). 0.2195g of KH2PO4 was dissolved in distilled
water up to the mark of a liter volumetric flask. This solution contains
50ppm phosphorus. A series of 0, 5,10,15,20 and 25ppm concentration
were prepared as follows:

26
0 ml, 5, 10,20and 25ml aliquots of the 50ppm standard solution were
prepared into a series of 50ml volumetric flasks of 50ml capacities. Two
ml of ammonium vanadate reagent were added to each of the volumetric
flasks. A yellow color is developed, usually after 30 minutes, the density
of color was measured using.

A standard curve was drawn by plotting the concentrations (ppm) against

corresponding readings of spectrophotometer. A straight line is
incidentally obtained.

3.3 Amino acid Determination

-Sample preparation for Hydrolysates Hydrolysates were prepared from

as following:

-200 mg of okra was weight in a hydrolysis tube to prepare Hydrolysate

sample.
-5 ml of 6 N HCl were added and the tube was tightly closed.
-The sample was incubated for 24 hours at 110 C.
-The sample was allowed to cool.
-125 mm filter paper was then use to filter the Hydrolysate.
-200 µl of the filtrate was taken into another tube and was evaporated
at 140 C ovens for about 1 hour. 1 ml of diluting buffer was then
added to the dried sample and transported to amino acid analyzer vial
and then Injected for analysis. After the separation of the injected
sample with a temperated cation separation column, Ninhydrine is
added continuously to the system. An integrated reagent dosing pump
is responsible for the delivery of this reagent, while an external buffer
pump is delivering the eluent. After the adding of Ninhydrine, the
eluent is lead through a high temperature reactor coil of about 16 m

27
length. With a typical flow rate of 0.7 m1/min the buffer/ninhydrine
mixture stays in the heated reactor for about 2 min. During this time
the mixture is heated to 130o C to accelerate the chemical reaction
outlined below:

28
The reaction products of the primary Amino Acids have a maximum
absorption at 570 nm, while the Secondary Amino Acids have their
maximum at 440 nm. After the reaction the mixture is lead through a
dual-channel photometer where both wavelengths (570 nm and 440 nm)
are measured continuously.

3.4 In vitro protein digestibility determination

Determination of protein digestibility of each sample was carried out
according to Sounders et al (1973) method. In a100 ml conical flask,
about 250 mg of material was suspended in 15 ml of 0.IN HCl contained
1.5 mg pepsin (1:10,000) and incubated at 37 C° for 3hr. the solution was
neutralized with 0.5N NaOH and treated with 4 mg pancreatin (Grade VI
porcine) in 7.5 ml of 0.2M phosphate buffer (pH 8.0) containing 0.005M
sodium azide. The mixture was incubated at 37 C° for 24hr. ten milliliters
10% trichloroacetic acid (TCA) was added to the mixture to stop the
reaction. The mixture was then centrifuged at 5000 rpm for 5 minutes.
Five milliliters a liquates from the supernatant were taken and analyzed
for nitrogen using semi microkjeldahl procedure (AOAC, 1984). The in
vitro protein digestibility was calculated using the formula:

- PD (%) = (N1-N2) X10

N3

- Where:
- PD = protein digested
- N1 = nitrogen in supernatant
- N2 = enzyme nitrogen
- N3 = total nitrogen

29
3.5 Anti-nutritional factors determination

3.5.1 Determination of phytic acid

The phytic acid content was determined according to themethod of

Wheeler and Ferrel (1971).

Two grams of sample were weighed in 125 conical flasks. The sample
was extracted with 40ml of 3% Trichloroacetic acid (TCA) for 3 hr. with
mechanical shaking. The suspension was centrifuged for 5 min. at 2500
rpm.

Ten milliliters aliquot of the supernatant were transferred to 40 ml

tube; and then 4 ml of FeCl3 (solution containing 2mg ferric iron (Fe+3)
per ml 3% TCA) were added to the aliquot. The tube was heated in a
boiling water bath for 45min. one or two drops of 3% Na2 SO4 in 3%
TCA were added to develop a precipitate. The tube was cooled and
centrifuged for 10-15min. at 2500 rpm. The clear supernatant was
decanted and the precipitate was washed twice by dispersing well in 25ml
3% TCA, heated for 10-15min in boiling water bath, then cooled and
centrifuged. The precipitate was washed one or two times with water, and
was dispersed in a few ml of water. Three milliliters of 1.5N NaOH were
then added and the volume completed to 30 ml with distilled water. The
tube was heated in a boiling water bath for 30min, and hot filtered using
Whatman No. 1 filter paper. The precipitate was washed with hot 60-
70ml of water and washings were decanted. The precipitate was dissolved
from the filter paper with 40 ml hot 3.2N HNO3 into 100 ml volumetric
flask and the paper was washed again with water in the same flask and
completed to volume with water.

30
 A volume of 0.5ml of the above suspension was transferred into 10 ml
volumetric flask. Two milliliters of 1.5N KSCN were added and
completed to volume with water, then immediately the absorbance was
read using spectrometer (JENWAY 6305 UV) at 480 nm.

A standard curve of different (Fe (NO3)2 Concentration was plotted to

calculate the ferric ion concentration. The phytate phosphorus was
calculated from the ferric ion concentration assuming 4:6 iron:
phosphorus molar ratio.

Calculation:

Phytate (mg/100g) = ( 6 /4 ) AX C X 20 X 10 X 50 X 100

1000 X S

Where:

-A = optical density

-C = Concentration corresponding to the optical density

-S = weight of sample

3.5.2 Determination of tannin content

Tannin content (TC) of pearl millet samples were estimated using

modified Vanillin-HCl in methanol as described by price et al (1978).
About 0.2g of ground sample was placed in 100ml conical flask. Ten
milliliters of 1% HCL in methanol (v/v.) were added, the contents were
mechanical shaking for 20 minutes and centrifuged at 2500pm for
5minutes. One ml of supernatant was pipettes into a test tube and 5
milliliters of vanillin-HCl reagent (mixing equal volume of 8%
concentrated HCl in methanol and 1% vanillin methanol) were added.
31
The optical density was read using a colorimeter (Lab System Analyzer-
9filters, J, Mitra and Bros. Pvt. Ltd). At 500nm after 20 minutes
incubation at 30C, a blank sample was carried out with each run of
samples. A standard curved was prepared expressing the result as
catechin equivalents, i.e. Amount of catechin (mg per ml) which gives
color intensity equivalent to that given by tannin after correcting for
blank.

Quantitative estimation of tannins was carried out using the reagent

prepared just at need to use by mixing equal volumes of 1% vanillin
methanol and 8% HCl methanol. It was discarded if a trace of color
appeared.

3.5.2.1 Tannin standard curve

Catechin was used to prepare the standard curve. This was done by
adding 600mg of D (+) catching to 100 ml of 1% HCl methanol. From
this stock solution various dilutions were prepared. Five ml of vanillin-
HCl reagent (0.5%) were added to 1ml of each dilution. The absorbance
was read using spectrophotometer (HENWAY 6305 UV/Vis.) at 500nm
after 20min. from addition of reagent at 30C. the absorbance was plotted
against catechin concentration Fig (6).

Procedure:

A weight of 0.2g of the samples was placed in a test tube. Then 10ml
of 1% HCl/ methanol were added. The test tube was capped and
continuously shaken for 20min. and then centrifuged at 2500 rpm for
5min. one ml of the supernatant was pippeted into each of the tubes and
then proceeding as was described in the standard cruve above.

32
 For zero setting prior to absorbance was read, 1ml blank solution
was mixed with 5ml 4% HCl/ methanol and 5ml vanillin reagent in a test
tub. The sample and blank test tubes were incubated for 20min. at 30C.

Absorbance was read determined from standard curve. Tannin

concentration was expressed as % as follows:

- CE (%) = C X 10 X100

200

Where:

- C= Concentration corresponding to the optical density

-10 = Volume of extract (ml)

- 200= sample weight (mg)

3.5.3 Total polyphenol (TP) determination:

Ployphenolic of each sample was estimated using Prussian blue assay,

as described by Price and Butter (1977). About 60mg of ground sample
extracted with 3ml methanol in a 50ml conical flask, and then poured into
a filter paper. The tube was quickly rinsed with additional 3ml methanol
and the content poured once into the filter paper. The filtrate was diluted
to 50ml with distilled water, mixed with 3ml 0.1M FeCL3 in 0.IN HCl
for 3 minutes, followed by the timed addition of 3ml 0.008M K3
Fe(CN)6. The absorption was read after 10 minutes at 720 nm on
spectrophotometer (corning, 259).

3.5.3.1 Standard curve preparation

33
Tannic acid standard curve was prepared by dissolved 100mg tannic acid
in distilled water in a. Liter volumetric flask and mad up to mark. This
prepared stock solution of 100ppm. Various standard concentrations (0, 2,
4,6, 8 and 10) were prepared. The Prussian blue assay described above
was then employed to the standard solution. The standard curve was
obtained by plotting concentration agnasit the corresponding absorbance
reading, which gave linear relationship.

Total polyphenol (%) = CX56 X100

60

Where:

-C= concentration corresponding to optical density

-56= Total volume

-60= weight of sample in milligrams

34
 CHAPTER FOUR

RESULTS AND DISCUSSION

4.1 Chemical composition

4.1.1 Moisture Content

With regard to the data shown in Table (1) the moisture content of
fresh okra was 83.6 % and 6.13 for dried okra. The moisture content
decreased through drying due to evaporation of water. The Result of this
study for fresh samples was slightly higher than that reported by Yousif
(1993) who gave a value 76.95%. Generally, dried okra had way less
moisture content than fresh okra (P≤.05)

4.1.2 Protein content

The protein content of fresh and dried Okra is preserved on table (1)
dried Okra had significantly (P≤0.5) higher protein Values than the fresh
Okra. Obviously such an increase in the protein value is due to the
concentration effect of drying. The value reported in this study were
slightly less than that reported by FAO (1970) who gave a value of 14.2%
but way higher than that reported by Boutros (1977) who reported a value
of 2.3g/100g .

4.1.3 Fat content

As illustrated in table (1) it is clear that fat content of fresh and

dried okra were significantly different (P≤0.5) fat content of dried okra
were (2.7%) were considerably higher than that for fresh Okra (1.8%) .
Boutros (1977) reported that fat content of Okra 0.1g/100g.

35
Table 1: Chemical Composition of Okra:

Content Fresh okra Dried okra

a b
Moisture (%) 83.6 ± 0.78 6.13 ± 0.15

D.M (%) 16.37a ±0.26 b

93.6 ±0.15
a b
Ash (%) 2.0 ±0.6 7.9 ±0.15
a b
C.P (%) 11.3 ± 0.15 13.7 ±0.21
a b
Fat (%) 1.8 ±0.06 2.7 ±0.10
a b
C.F (%) 28.6 ±0.31 34.5 ±0.42
Note:

*n=3

*a –b Means in the same row bearing different superscript letters are

significantly different at (P≤0.05)

36
4.1.4 Ash content

According to the results summarized in table (1) apparently dried okra

has way higher ash content than fresh okra (P≤.05).

4.1.5 Fiber content

The result of crude fiber content of the fresh and dried okra are shown
in Table (1) it is clear that fiber content of dried okra (34.5%) was higher
than that for fresh one (28.6%). The fiber content of (Hibiscus esculentus)
is 1.6 g/100g reported by Botrus (1977). Moreover Eduard and Miller
(1947). Analyzed samples of okra seed meal and found that the fiber
8.14%.

4.2 Minerals composition

The effect of drying on the minerals content of okra pods are shown
in Table (2).

From table (2) it can be seen that the sodium content of the dried okra
(0.09%) was found to be higher than that of fresh okra (0.07%), (P≤0.05).
There was significant difference (P≤0.05). In the Potassium contents
between fresh and dried okra while that Potassium of fresh okra was
higher than that data reported by Ahmad (1968) and lower than that
obtained by Yousif, (1993).This variation may be due to the variety
difference or type of soil . Clearly the calcium content of the dried okra is
higher than that of fresh okra table (2). The value of calcium content of
fresh okra was found to be lower than that reported by Yousif,N.E.
(1993) . There was significant increases (P≤0.05) in calcium content

37
thought the drying. Considerable differences (P≤0.05) in Phosphorous

Content Na P Ca K

a a a a
Fresh okra 0.07 ± 003 0.26 ±.05 0.36 ±.05 0.57 ±.05

a b a b
Dried okra 0.09 ±.003 0.32 ±.01 0.38 ±.05 0.66 ±01

content were observed between the fresh and dried Okra.

Table 2: minerals content okra:

Note:

*n=3

*a –b Means in the same column bearing different superscript letters are

significantly different at (P≤0.05) .

38
4.3 Amino Acids profile

The Effect of drying on the amino acids profile on okra (hibiscus

esculentus) is displayed on table (3) on the average fresh Okra contained
atotal of 1918.09 mg\100g of Okra pods Expectedly drying result in an
obviously (P<0.001) increases in all amino acids. Table (3) Increases in
the range of 337.4% To 438.3 % are reported. The highest and the least
amino acids in Okra (hibiscus esculentus) wither in weight wet basis or
dry weight basis are reported for Aspartic acid and Methionine
respectively. the ratio of essential to non essential Amino Acids
(ESAA\NEAA) range between a heights of 0.92 and a low of 0.49 for the
dried and fresh Okra (hibiscus esculentus ) respectively. El Gasim and Al
Kanhal (1992). Reported that the ratio of ESAA \NEAA of Beef to be
0.85 and for goat 0.9, on dry weight basis Okra hade ESAA \NEAA
similar to that of beef or goat .on weight basis Okra content an
appreciable amount of Lysine (113.1mg\100g ) its well Known that
Lysine is the limiting amino acid in Cereals ( Bressani et al .1963)
Kisraa is one of popular dishes in Sudan .Kisraa normally etan with Okra
Molah since Kisraa made from cereal can complement Kisra

Okra as a potential high –protein source witch due to its high Lysine
level and most limiting amino acids were found may serve as a
supplement to cereal based diets.

39
Table (3): Amino acid profile of Okra

Type of Fresh okra Dried okra

Amino Acid (mg/100g) (mg/100g)

Aspartic acid 394.70a ±66.02 2124.59b ±219.57

Threonine 77.57a ±15.08 295.53b ±35.72

Serine 68.48a ±10.18 202.25b ±19.66

Glutamic acid 163.68a ±25.05 163.68b ±25.05

Glycine 57.86a ±7.41 144.43b ±34.49

Alanine 141.15a ±31.13 563.03b ±64.65

Valine 121.53a ±27.53 547.08b ±64.97

Methionine 35.47a ±10.45 155.15b ±39.41

Isoleucine 90.46a ±20.19 388.56b ±48.17

Leucine 133.93a ±28.62 574.14b ±67.66

Tyrosine 46.43a ±10.81 131.58b±24.58

Phenylalanine 81.12a ±17.41 326.28b ±46.23

Histidine 76.80a ±14.37 322.15b ±38.30

Lysine 113.08a ±16.94 346.98b ±67.51

Arginine 119.77a ±22.08 704.62b ±90.36

Ammonia 250.59a ±32.15 1067.57b ±51.65

40
Note: * n = 3

*a –b Means in the same row bearing different superscript letters are

significantly different at (P≤0.001).

4.4 Protein Digestibility

Effect of drying on the invitro protein digestibility of Okra(hibiscus

esculentus) is preserved on Table (4) In vitro protein digestibility of
fresh Okra was quite low only 30.72 % Table (4) Drying Improved
greatly the in vitro protein digestibility and raised it to 82.37 % . Such an
Improvement in in vitro protein digestibility will improve the nutritive
value of Okra our knowledge's unfortunately no data is reported on the
effect of drying on the in vitro protein digestibility of Okra (hibiscus
esculentus)

4.5 Anti-nutritional Factors

When effect of drying on the anti-nutritional factors ( tannin ,

polyphenol, phytic ) of Okra hibiscus esculentusis presented in table (4)
The means concentration of tannin and phytic acid of dried Okra were
found to be significantly lower (P<.05) than that of fresh Okra Table (4).
However the extent of reduction in phytic acid was found to be way less
than that for tannin .Like wise polyphenols were greatly reduced by about
21.7%. polyphenols in fresh and dried Okra were 706.2 mg\100g and 533
mg\100g for dried Okra respectivel

41
Table (3): Protein Digestibility Of fresh and Dried Okra

Type Protein digestibility P% ± SD

Fresh b
30.72 ± 4.54

Dried 82.57a ±3.57

Note:
*n=3

*a –b Means in the same row bearing different superscript letters are

significantly different at (P≤0.001).

42
Table (5): Determination of anti nutritional factors in okra
mg/100g

Factor Tannin Polyphenol Phytic Acid

mg/100g mg/100g mg/100g

Type
Fresh a a a
74.2 ±8.60 706.2 ±47.73 33.96 ±1.4

Dried b b b
22.55 ±3.81 553 `±24.95 33.09 ±0.37

*n=3

*a –b Means in the same row bearing different superscript letters are

significantly different at (P≤0.001).

43
 CHAPTER FIVE

CONCOLUSIONS AND RECOMMENDATIONS

CONCOLUSIONS:

* Tradition sun drying method applied in the present investigation

resulted in a reduction of anti nutritional factors (phytic acid, tannin and
polyphenol).
*Quantitative reduction in anti nutritional factors was found to be
accompanied by improvement in protein and minerals availability as
evidenced by in vitro protein digestibility and minerals, respectively.
*The results indicated that tradition sun drying of Okra is a potential
process to produce an improved Okra diet with high nutritive value.

RECOMMENDATION:

* Tradition Sun Drying could be a useful means to improve Okra for

human nutrition.

* Further investigations are needed to optimize the processing techniques

for further improvement in protein digestibility and mineral extractability
by reducing the anti nutritional factors.

*Drying techniques could be good methods for preservation of okra.

44
* Okra as a potential high –protein source witch due to its high Lysine
level and most limiting amino acids were found may serve as a
supplement to cereal based diets.

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52