Effect of processing on Chemical Composition and

Transcription

Effect of processing on Chemical Composition
and biological value of cowpeas
By
Gaafar Abdelatif Nugdallah
B.Sc. (Agric.), Al Azhar University – 1977
M.Sc. (Agric.) University of Khartoum - 1996
Supervisor: Prof. Abdullahi Hamid El Tinay
A Thesis
Submitted to the University of Khartoum in fulfillment for the
requirements of the degree of Doctor of Philosophy (Agric.)
Department of Food Science and Technology
Faculty of Agriculture, University of Khartoum
October – 2003
1
DEDICATION
This humble effort is
dedicated with all love and
gratitude to our prophet
Mohamed (Peace and Blessing be
upon him) through whom Allah
the Al Mighty says in Glorious
Quran.
In the name of Allah, the
Beneficent the Merciful
‫ﺑﺴﻢ اﷲ اﻟﺮﺣﻤﻦ اﻟﺮﺣﻴﻢ‬
24. Then let man look at his
food, (And how We provide
it)
25. For that We pour forth
Water in Abundance,
26. And We split the earth In
fragments,
27. And produce therein
grain,
28. And Grapes and The fresh
vegetation,
29. And Olives and Dates,
30. And enclosed Gardens,
Dense with lofty trees,
31. And fruits and Fodder,
32. A provision For you and
2
(24) ‫ﻓﻠﻴﻨﻈﺮ ﺍﻹﻧﺴﺎﻥ ﺇﱃ ﻃﻌﺎﻣﻪ‬
(25) ‫ﺃﻧﺎ ﺻﺒﺒﻨﺎ ﺍﳌﺎﺀ ﺻﺒﹰﺎ‬
(26) ‫ﰒ ﺷﻘﻘﻨﺎ ﺍﻷﺭﺽ ﺷﻘﺎ‬
(27) ‫ﻓﺄﻧﺒﺘﻨﺎ ﻓﻴﻬﺎ ﺣﺒﺎ‬
(28) ‫ﻭﻋﻨﺒﹰﺎ ﻭﻗﻀﺒﺎ‬
(29) ‫ﻭﺯﻳﺘﻮﻧﹰﺎ ﻭﳔﻼ‬
(30) ‫ﻭﺣﺪﺍﺋﻖ ﻏﻠﺒﺎ‬
(31) ‫ﻭﻓﺎﻛﻬﺔ ﻭﺃﺑﺎ‬
(32) ‫ﻣﺘﺎﻋﹰﺎ ﻟﻜﻢ ﻭﻷﻧﻌﺎﻣﻜﻢ‬
your cattle
Sura Abasa, Ayat (24 – 32).
ZttytÜ
TABLE OF CONTENTS
Page
Dedication..................................................................................................................................
ii
List of Contents …...............................................................................................................
iii
List of Tables............................................................................................................................
viii
List of Structure .....................................................................................................................
ix
List of Figures ….....................................................................................................................
x
Acknowledgement ................................................................................................................. xi
Abstract .....................................................................................................................................
xii
Arabic Abstract
xx
..................................................................................................................
CHAPTER ONE: INTRODUCTION ..................................................................... 1
1.1. Importance of cowpea.................................................................................................
2
1.2. Classification of cowpea............................................................................................. 4
1.3. Antinutritional factors.................................................................................................
5
1.4 Processing of cowpeas.................................................................................................. 7
1.5 Utilization of cowpea....................................................................................................
8
1.6. Production of cowpea................................................................................................
10
3
Objective of the study .........................................................................................................
11
CHAPTER TWO: LITERATURE
REVIEW........................................
12
2.1. Nutritional value of cow pea
..................................................................................
13
2.2. Protein fractionation....................................................................................................
16
2.2.1. Nitrogen solubility (NS) ........................................................................................
16
2.2.2. Protein fractions classification............................................................................
17
2.2.2.1 Albumins and globulins.....................................................................................
19
2.2.2.2. Prolamin..................................................................................................................... 20
2.2.2.3 Glutelin.......................................................................................................................
21
2.2.2.3.2. G1 - glutelin.........................................................................................................
22
2.2.2.3.2. G2 - glutelin.........................................................................................................
22
2.2.2.3.3 G3 - glutelin..........................................................................................................
23
2.2.2.4. Insoluble protein...................................................................................................
24
2.3 Anti-nutritional factors.................................................................................................
25
2.3.1. Chemical nature.........................................................................................................
26
2.3.1.1. Chemical nature of tannin.................................................................................
26
2.3.1.2. Chemical nature of phytic acid.......................................................................
30
2.3.1.3 Chemical nature of the inhibitors....................................................................
33
4
2.3.2 Anti-nutritional effect.....................................................................................
33
2.3.2.1 Anti-nutritional effect of the Inhibitors........................................................ 33
2.3.2.2 Anti-nutritional effect of the tannin................................................................ 36
2.3.2.3 Anti-nutritional effect of phytic acid............................................................
38
2.3.3 Anti-nutrients content ..............................................................................................
41
2.3.3.1 Tannin content of cowpea.................................................................................... 41
2.3.3.2 Trypsin inhibitor activity.....................................................................................
44
2.3.3.3 Content of phytic acid.........................................................................................
47
2.4. Processing........................................................................................................................
48
CHAPTER THREE: MATERIALS AND METHODS………………......…
64
3.1 Material..............................................................................................................................
64
3.1.1 Food materials..............................................................................................................
64
3.1.1.1 Chemical and reagents.....................................................................................
64
3.1.2 Apparatus.......................................................................................................................
64
3.2 Methods..............................................................................................................................
65
3.2.1 Preparation of cowpea samples...........................................................................
65
3.2.1.1 Cleaning......................................................................................................................
65
3.2.1.2 Autoclaving................................................................................................................ 65
3.2.1.3 Roasting....................................................................................................................... 66
3.2.1.4 Germination of cowpea samples.....................................................................
5
66
3.2.1.5 Cooking of sprouted cowpea samples............................................................ 67
3.3 Analytical methods......................................................................................................... 67
3.3.2 Protein fractionation..................................................................................................
67
2.3.3. In-vitro protein digestibility IVPD....................................................................
71
3.3.4 Determination of tannin in raw and treated samples................................
74
3.3.5 Determination phytic acid.....................................................................................
77
3.3.6 Determination of trypsin inhibitory factor....................................................... 81
3.3.7 Statistical analysis....................................................................................................... 82
CHAPTER FOUR: RESULTS AND DISCUSSION………………..……......… 83
4.1 Proximate composition................................................................................................
84
4.2 Protein fraction................................................................................................................
86
4.1.3. Ash content................................................................................................................... 87
4.1.4. Fiber content................................................................................................................
88
4.1.5. Oil content...................................................................................................................
89
4.2. Protein fraction ..............................................................................................................
89
4.2.1 Globulin fractions........................................................................................................ 89
4.2.2 Albumin fraction........................................................................................................
90
4.2.3. Prolamin fraction........................................................................................................ 93
4.2.4 G1-glutelin fraction..................................................................................................... 93
4.2.5 G2-glutelin fraction..................................................................................................... 94
6
4.2.6. G3-glutelin fraction.................................................................................................... 95
4.2.7 Residue fraction........................................................................................................... 96
4.3. Anti-nutrients ................................................................................................................... 101
4.3.1. Tannins............................................................................................................................ 101
4.3.2. Trypsin inhibitor activity.......................................................................................
102
4.3.3. Phytic acid..................................................................................................................... 102
4.3.4 In-vitro protein digestibility IVPD...................................................................... 104
CHAPTER FIVE SUMMARY, CONCLUSION AND
RECOMMENDATIONS…………………………………………………………….……......
111
Recommendation........................................................................................................................
113
REFERENCES……………..…………….………………………….……......…………………...... 114
7
LIST OF TABLES
Table Title
No.
1.
Protein extraction Procedure for sequence Ao and Bo.................
70
2.
Proximate composition of some cowpea preparations................... 85
3.
Effect of germination on protein fractions of cowpea cultivars
4.
Effect of cooking on protein fractions of germinated cowpea
cultivars...............................................................................................................
5.
98
99
Phytic acid (mg/100g dry weight), Tannins (g/100g dry
weight), Trypsin inhibitor activity (TUI/mg protein) of raw
and processed cowpea varieties and IVPD.........................................
6.
100
Effect of varying concentration of treated sample (roasting,
autoclaving) + raw cowpea seed flour on IVPD............................... 106
7.
Effect of autoclaving and roasting on protein fractions of
cowpea cultivars............. ............. ............. ............. ............. ............. ..........
8
110
LIST OF STRUCTURES
Struc. Title
No.
1.
Glalotannin......................................................................................................... 28
2.
Gallic acid........................................................................................................... 28
3.
Hexahydroxy-diphenic acid (Ellagitannin) ......................................... 28
4.
Ellagic acid (Dilact) ......................................................................................
5.
Flavanzol (Catechin) ..................................................................................... 29
6.
Flavan 3-4-diol (leucoantho-cyanidin) ................................................
7.
3-Hydroxy flavylium (Anthocyanidin) ................................................. 29
8.
Apossible structure of grain sorghum condensed tannin............... 29
I
Myo – inositol.....................................................................................................
32
II
Phytate ions..........................................................................................................
32
III
Phytate ions..........................................................................................................
32
IV
Polymeric structutre..........................................................................................
32
V
Phytate ions.........................................................................................................
32
9
28
29
LIST OF FIGURES
Fig.
1
2
Page
Catchen concentration mg/ml relationship between optical
density at 500nm and catechin concentration.................................
76
Phytic acid standard curve ................... ................... ................... ..........
80
10
Acknowledgements
I wish to express my deep sense of gratitude and sincere
thanks to my supervisor A.H. El Tinay for his continuous interest
helpful, guidance, encouragement, criticisms, advice and
supervision throughout the progress of this work.
My sincere thanks and due to my colleagues and technical
staff and workers of the Department of Food Science and
Technology for their assistance, in particular to Mr. El Habib and
Ms. Asma, Mr. Abass and Madam Ihsan.
Best regards are due to Mr. Abdelhamed for his unfailing
patience and skill in typing the manuscript.
I am grateful to my family members who, with great
patience, tolerated all the inconveniences resulting and for being
of help in this work that no words can express my feelings
towards them.
Lastly, my thanks are extended to others who offered help
in one way or another.
Above all my special praise and unlimited thanks are to
Allah, who helped me and gave me health and patience to
complete this study.
11
ABSTRACT
Two cowpea cultivar (Vigna unguiculata) namely "Ain Elgazal"
and "Buff" were obtained from Elobeid Research Station and were
used in this study.
The seeds were analyzed for their chemical composition,
protein soluble fractions, in-vitro protein digestibility (IVPD), phytic
acid, trypsin inhibitor activity and tannin content of raw, boiled,
roasted, autoclaved, germinated for four days, and germinated cooked
samples.
The proximate composition showed that moisture content
ranged from 4.4% to 5.9% for Ain Elgazal cultivar and ranged from
4.4% to 5.5% for Buff cultivar. Protein content ranged from 25.6% to
31.0% for Ain Elgazal cultivar and ranged from 24.6% to 28.9% for
Buff cultivar. Ash content ranged from 3.2% to 4.3% for Ain Elgazal
cultivar and ranged from 3.5% to 4.3% for Buff cultivar. Fibre content
ranged from 2.5% to 3.2% for Ain Elgazal cultivar and ranged from
2.2% to 3.1% for Buff cultivar. Oil content ranged from 1.5% to 1.6%
for Ain Elgazal cultivar and ranged from 1.6% to 1.7% for Buff
cultivar. All the processing treatments had little effect on oil content
and significantly (P ≤ 0.05) increased protein content, ash content and
fibre content for both cultivars.
The raw and germinated samples were fractioned, the major
protein fraction Globulin showed significant (P ≤ 0.05) decrease, they
were 87.5%, 80.6%, 76.4%, 71.4% and 70.2% for Ain Elgazal raw
seeds, the first, second, third and fourth days respectively. Similarly
for buff raw and germinated seeds, they were 89.8, 81.3, 78.3, 75.6
and 72.7% respectively. The Albumin fraction significantly (P < 0.05)
12
decreased they were 4.0%, 3.8%, 2.3%, 1.9% and 2.5% for Ain
Elgazal raw seeds, the first, second, third and fourth days respectively.
Similarly for Buff raw and germinated seeds, 3.6%, 1.5%,
2.2%, 2.2% and 1.2% respectively. The prolamin fraction for Ain
Elgazal raw and germinated seeds were 4.0%, 3.3%, 4.0%, 1.6% and
4.6% respectively. Similarly for buff raw and germinated seeds were
4.5%, 2.7%, 2.8%, 3.9% and 1.4% respectively. The G1-glutelin
fraction for Ain Elgazal raw and germinated seeds showed significant
(P ≤ 0.05) decrease, they were 2.4%, 2.1%, 1.3%, 0.8% and 1.7%
respectively. Similarly for Buff raw and germinated seeds were 2.3%,
1.2%, 1.3%, 2.1% and 1.3% respectively. The G2-glutelin fraction for
Ain Elgazal raw and germinated seeds were 2.4%, 2.1%, 1.9%, 1.4%
and 1.7% respectively. For Buff raw and germinated seeds were 2.5%,
6.4%, 5.1%, 3.9% and 4.5% respectively. The G3-glutelin fraction
showed significantly (P ≤ 0.05) increase, they were 4.8%, 9.8%,
12.0%, 13.5% and 10.4% for Ain Elgazal raw seeds, the first, second,
third and fourth days respectively. Similarly for Buff, they were 4.5%,
10.0%, 12.5%, 11.1% and 11.7% respectively. The insoluble protein
fraction showed significant (P ≤ 0.05) increase, they were 1.2%, 2.5%,
2.1%, 9.2% and 6.0% for Ain Elgazal raw seeds, the first, second,
third and fourth days respectively. Similarly for Buff raw and
germinated seeds were 1.2%, 1.7%, 1.9%, 3.6% and 6.1%
respectively.
Cooking significantly (P ≤ 0.05) reduced globulin, they were
16.4%, 19.0%, 18.1%, 18.0% and 17.8% for Ain Elgazal raw first,
second, third and fourth days respectively. Similarly for Buff raw and
13
respectively. Cooking significantly (P ≤ 0.05) reduced Albumin, they
were 1.9%, 2.8%, 3.1%, 1.8% and 1.4% for Ain Elgazal raw, first,
second, third and fourth days respectively. Similarly for Buff raw and
respectively. Cooking significantly (P ≤ 0.05) increased prolamin
fraction, they were 4.2%, 6.3%, 4.8%, 5.1%, and 5.0% for Ain
Elgazal raw, first, second, third and fourth days respectively.
Similarly for Buff raw and first and then decreased second, third and
fourth days they were 4.6%, 5.1%, 2.7%, 2.7% and 1.8% respectively.
Cooking significantly (P ≤ 0.05) decreased G1-glutelin fraction
they were 2.0%, 1.4%, 1.3%, and 1.3% for Ain Elgazal raw, first,
second, and third days respectively, but it increased to 2.6% in the
fourth day. Similarly for Buff raw and germinated seeds were 2.7%,
1.2%, 1.1%, 1.6% and 1.0% respectively. Cooking significantly (P ≤
0.05) increased G2-glutelin fraction, they were 3.4%, 4.4%, 4.4%,
4.0% and 5.7% for Ain Elgazal raw, first, second, third and fourth
days respectively. Similarly for Buff raw and germinated seeds were
5.1%, 7.5%, 7.5%, 7.9% and 8.8% respectively.
Cooking significantly (P ≤ 0.05) increased G3-glutelin fraction,
they were 63.8%, 61.0%, 62.0%, 61.6% and 57.0% for Ain Elgazal
raw first, second, third and fourth days respectively. Similarly for Buff
raw and germinated seeds were 69.3%, 59.0%, 62.3%, 60.1% and
62.1% respectively. Cooking significantly (P ≤ 0.05) increased the
insoluble protein fraction, they were 5.5%, 8.0%, 8.8%, 9.6% and
10.2% for Ain Elgazal raw, first, second, third and fourth days
respectively. Similarly for Buff raw and germinated seeds were 5.5%,
6.6%, 6.5%, 6.1% and 6.5% respectively.
14
Autoclaving at 120ºC under 15 psi for 30 min for Ain Elgazal
raw seeds significantly (P ≤ 0.05) reduce globulin fraction from 87.5%
to 29.2% and albumin from 4.0% to 1.2% but increased prolamin from
4.3% to 4.5%, reduced G1-glutelin from 2.3% to 1.2% and G2-glutelin
from 2.4% to 1.2% and increased G3-glutelin from 4.8% to 59.5% and
insoluble protein fraction from 1.2% to 8.0%.
Autoclaving at 150ºC under 20 psi for 30 min for Ain Elgazal
raw seeds significantly (P ≤ 0.05) reduce globulin fraction from 87.5%
to 27.6%, albumin fraction from 4.0% to 1.2% but increased prolamin
from 4.3% to 4.8%, decreased G1-glutelin from 2.3% to 2.0%,
increased G2-glutelin from 2.4% to 4.0%, increased G3-glutelin from
4.8% to 60.2% and increased insoluble protein fraction from 1.2% to
6.0%.
Roasting Ain Elgazal raw seeds at 90ºC for 60 min significantly
(P ≤ 0.05) reduce globulin fraction from 87.5% to 85.0%, albumin
showed changeless, prolamin fraction reduced from 4.3% to 3.6%,
G1-glutelin fraction reduced from 2.3% to 2.0%, G2-glutelin fraction
reduced from 2.4% to 1.2%, G3-glutelin fraction increased from 4.8%
to 6.0% and insoluble protein fraction increased from 1.2% to 3.0%.
Roasting Ain Elgazal raw seeds at 120ºC for 60 min significantly (P ≤
0.05) reduced globulin fraction from 87.5% to 41.0%, albumin
fraction reduced from 4.0% to 1.2%, prolamin fraction reduced from
4.3% to 4.2%, G1-glutelin fraction reduced from 2.3% to 1.2%, G2glutelin fraction increased from 2.4% to 3.2%, G3-glutelin fraction
increased from 4.8% to 48.0% and insoluble protein fraction increased
from 1.2% to 6.0%. Autoclaving at 120ºC under 15 psi for 30 min for
Buff raw seeds significantly (P ≤ 0.05) reduced globulin fraction from
15
89.8% to 32.0%, albumin fraction from 3.6% to 1.0%, prolamin from
4.5% to 3.0%, decreased G1-glutelin from 2.3% to 2.0%, G2-glutelin
fraction increased from 2.5% to 3.0%, G3-glutelin increased from
4.5% to 55.0% and insoluble protein fraction increased from 1.2% to
6.0%.
Autoclaving at 150ºC under 20 psi for 30 min for Buff raw
seeds significantly (P ≤ 0.05) reduce globulin fraction from 89.5% to
36.0%, albumin fraction from 3.6% to 1.0% prolamin fraction from
4.5% to 4.0%, G1-glutelin from 2.3% to 1.2%, G2-glutelin increased
from 2.5% to 3.2%, G3-glutelin increased from 4.5% to 50.8% and
increased insoluble protein fraction from 1.2% to 7.0%. Roasting Buff
raw seeds at 90ºC for 60 min significantly (P ≤ 0.05) decreased
globulin fraction from 89.5% to 88.0%, albumin showed changeless,
prolamin fraction decreased from 4.5% to 4.0%, G1-glutelin from
2.3% to 2.2%, G2-glutelin fraction increased from 2.5% to 2.8%, G3glutelin fraction showed changeless, insoluble protein fraction
increased from 1.2% to 1.4%. Roasting Buff raw seeds at 120ºC for 60
min significantly (P ≤ 0.05) decreased globulin fraction from 89.8% to
45.5%, albumin fraction from 3.6% to 1.0%, prolamin fraction
increased from 4.5% to 6.0%, G1-glutelin decreased from 2.3% to
2.0%, G2-glutelin fraction increased from 2.5% to 3.0%, G3-glutelin
fraction increased from 4.5% to 42.0% and insoluble protein fraction
increased from 1.2% to 6.0%.
Samples were analyzed for phytic acid, tannin and trypsin
inhibitor activity. Germination reduced phytic acid from 310.3
(mg/100g dry weight) of raw seeds to 286.1 mg/100g, 248.9 mg/100g,
201.7 and 139.8mg/100g for Ain Elgazal germinated seeds in the first,
16
second, third and fourth days respectively. Similarly for Buff raw
seeds phytic acid was 376.3 which reduced by germinated to 346.2,
301.0, 225.7 and 180.7mg/100g for the first, second, third and fourth
days respectively.
Cooking showed significant (P ≤ 0.05) decreased of phytic acid
for Ain Elgazal raw and germinated seeds, they were 290.3mg/100g,
268.9mg/100g, 229.0mg/100g, 181.5mg/100g and 128.6mg/100g for
raw, first, second, third and fourth days respectively. Similarly for
Buff they were 353.7mg/100g, 311.6mg/100g, 270.9mg/100g,
205.4mg/100g and 162.6mg/100g for raw, first, second, third and
fourth days respectively. Autoclaving at 150ºC under 20 psi for 30
min reduced phytic acid for Ain Elgazal raw seeds from 310.3 to
300.0mg/100g. Similarly for Buff raw seeds autoclaving reduced
phytic acid from 376.3 to 350.0mg/100g. Roasting at 120ºC for 60
min reduced phytic acid for Ain Elgazal raw seeds from 310.3 to
301.0mg/100g. Similarly for Buff roasting reduced phytic acid from
376.3 to 352.0mg/100g.
Germination significantly (P ≤ 0.05) decreased tannins from
0.48g/100g of Ain Elgazal raw seeds to 0.36g/100g, 0.30g/100g,
0.24g/100 and 0.20g/100g in the first, second, third and fourth days
respectively. Similarly for Buff raw seeds tannins was 0.50g/100g it
reduced by germination to 0.42g/100g, 0.36g/100g, 0.30g/100g and
0.22g/100g in the first, second, third and fourth days respectively.
Autoclaving at 150ºC under 20 psi for 30 min reduce tannins for
Ain Elgazal raw seeds from 0.48g/100g to 0.38g/100g. Similarly for
Buff autoclaving reduce tannins from 0.50g/100g to 0.40g/100g.
Roasting at 120ºC for 60 min reduced tannins for Ain Elgazal raw
17
seeds from 0.48g/100g to 0.36g/100g. Similarly for Buff roasting
reduced tannins from 0.50g/100g to 0.42g/100g.
Cooking showed significant (P ≤ 0.05) further reduction of
tannins for Ain Elgazal raw and germinated seeds, they were
0.26g/100g, 0.18g/100g, 0.15g/100g, 0.12g/100g and 0.10g/100g for
raw, first, second, third and fourth days respectively. Similarly for
Buff they were 0.30g/100g, 0.22g/100g, 0.20g/100g, 0.18g/100g and
0.11g/100g for raw, first, second, third and fourth days respectively.
Germination significantly (P ≤ 0.05) reduced trypsin inhibitor activity
for Ain Elgazal raw seeds from 22.0 TUI/mg protein of raw to 11.8
TUI/mg protein, 10.6 TUI/mg protein, 8.0 TUI/mg and 8.0 TUI/mg
protein for first, second, third and fourth days respectively. Similarly
for Buff they were 25.0 TUI/mg protein, 12.5 TUI/mg protein, 10.0
TUI/mg protein, 9.6 TUI/mg and 9.0 TUI/mg protein for raw, first,
second, third and fourth days respectively. Cooking autoclaving at
150ºC under 20 psi for 30 min and roasting at 120ºC for 60min
eliminates trypsin inhibitor activity.
Germination improved in-vitro protein digestibility (IVPD), it
were 73.4%, 75.3%, 77.9%, 80.4% and 84.4% for Ain Elgazal raw
seeds, first, second, third and fourth days, respectively. Similarly for
Buff they were 74.2%, 75.7%, 79.3%, 82.4% and 83.6% respectively.
Cookin germinating seeds showed significant (P < 0.05) further
increase of in-vitro protein digestibility, they were 86.2%, 87.2%,
87.5%, 88.8%, and 88.5% for Ain Elgazal raw, first, second, third and
fourth days respectively. Similarly for Buff were 85.4%, 86.3%,
86.6%, 87.9% and 88.3% for raw, first, second, third and fourth days
respectively. Autoclaving at 150ºC under 20 psi for 30min increased
18
IVPD for Ain Elgazal raw seeds from 73.4% to 86.0% and increased
by roasting at 120ºC for 60min to 84.0% Similarly Buff increased by
autoclaving from 74.2% to 87.0% and by roasting to 84.0%.
Effect of substrate on in-vitro protein digestibility was carried
out using mixed samples. Increasing the proportion of heat treated
material in comparison to raw material significantly (P ≤ 0.05)
increased in-vitro protein digestibility. For Ain Elgazal the proportion
were 0.192g raw materials plus 0.192 heat treated material, 0.096g
plus 0.282g, 0.048g plus 0.336g, 0.0 plus 0.384g, 0.288g plus 0.096g,
0.336g plus 0.048g and 0.384g plus 0.0g. The heat treated material
were roasted at 90ºC and 120ºC for 30min, 45min and 60min. The
material were autoclaved at 115.5ºC under 10psi and at 120ºCunder
15 psi for 15 min, 30 min and 45min. The IVPD ranged from 73.4%
to 84.0% and 73.4% to 84.2 for raw plus roasted mixed samples
respectively. The IVPD ranged from 73.4% to 82.0%, 73.4% to
83.0%, 73.4% to 84.0%, 73.4% to 85.0%, 73.4 to 85.5% and 73.4% to
86.2 for raw plus autoclaved (mixed samples) for Ain Elgazal
respectively.
Similarly for Buff IVPD were ranged from 74.2% to 79.7%,
74.2% to 79.9%, 74.2% to 79.8%, 74.2% to 83.8%, 74.2 to 84.0% and
74.2% to 84.6 for raw plus roasted (mixed samples) respectively. The
IVPD ranged from 74.2% to 80.2%, 74.2% to 84.8%, 74.2% to 84.8%,
74.2% to 85.0%, 74.2 to 85.0% and 74.2% to 85.4 for raw plus
autoclaved (mixed samples) respectively.
‫ﺑﺴﻢ اﷲ اﻟﺮﺣﻤﻦ اﻟﺮﺣﻴﻢ‬
19
‫ﺧﻼﺻﺔ ﺍﻷﻃﺮﻭﺣﺔ‬
‫ﺖ ﰲ ﻫﺬﻩ ﺍﻟﺪﺭﺍﺳﺔ ﺻﻨﻔﲔ ﻣﻦ ﺍﻟﻠﻮﺑﻴﺎ ﺍﻟﻄﻴﺐ ﺃﹸﺳﺘ‪‬ﺠﻠﺒﺖ ﻣﻦ ﻫﻴﺌﺔ ﺍﻟﺒﺤﻮﺙ ﺍﻟﺰﺭﺍﻋﻴﺔ ﳏﻄﺔ‬
‫ﺃﹸﺳﺘ‪‬ﺨﺪِﻣ ‪‬‬
‫ﺍﻷﺑﻴﺾ ﻭﻫﻲ ﻋﲔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﺍﻟﻌﻴﻨﺔ ﺑﻒ‪.‬‬
‫ﰎ ﻋﻤﻞ ﺍﻟﺘﺤﻠﻴﻞ ﺍﻟﺘﻘﺮﻳﱯ ﻟﻠﻌﻴﻨﺎﺕ ﻭﻛﺬﻟﻚ ﲡﺰﺋﺔ ﺍﻟﱪﻭﺗﲔ ﺣﺴﺐ ﺧﺎﺻﻴﺔ ﺍﻟﺬﻭﺑﺎﻥ ﻭﰎ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ‬
‫ﻣﻌﻤﻠﻴﹰﺎ ﻭﺗﻘﺪﻳﺮ ﲪﺾ ﺍﻟﻔﺎﻳﺘﻚ ﻭﻧﺸﺎﻁ ﻣﺜﺒﻂ ﺇﻧﺰﱘ ﺍﻟﺘﺮﺑﺴﲔ ﻭﺍﻟﺘﺎﻧﲔ ﻟﻠﻌﻴﻨﺎﺕ ﺍﳋﺎﻡ ﻭﺍﻟﱵ ﰎ ﺗﻮﺯﻳﻌﻬﺎ ﳌﺪﺓ ﺃﺭﺑﻌﺔ ﺃﻳﺎﻡ‬
‫ﻭﻟﻠﻤﻌﺎﻣﻠﺔ ﻃﺮﺩﻳﹰﺎ ﺑﺎﻟﻄﺒﺦ ﺍﻟﺘﻘﻠﻴﺪﻱ ﻭﺍﶈﻤﺼﺔ ﻭﺍﻟﻄﺒﺦ ﲢﺖ ﺿﻐﻂ ﺑﺎﻟﺒﺨﺎﺭ ﻭﺍﻟﻌﻴﻨﺎﺕ ﺍﻟﱵ ﰎ ﺗﺰﺭﻳﻌﻬﺎ ﰒ ﻃﺒﺨﺖ‪.‬‬
‫ﺃﻭﺿﺤﺖ ﻧﺘﺎﺋﺞ ﺍﻟﺘﺤﻠﻴﻞ ﺍﻟﺘﻘﺮﻳﱯ ﺃﻥ ﻧﺴﺒﺔ ﺍﻟﺮﻃﻮﺑﺔ ﺗﺮﺍﻭﺣﺖ ﺑﲔ ‪ %5.9 – 4.4‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﻭﺗﺮﺍﻭﺣﺖ ﺑﲔ ‪ %5.5-4.4‬ﻟﻠﻌﻴﻨﺔ ﺑﻒ‪ .‬ﻭﺗﺮﺍﻭﺣﺖ ﻧﺴﺒﺔ ﺍﻟﱪﻭﺗﲔ ﺑﲔ ‪ %30.0-25.6‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﻭﺗﺮﺍﻭﺣﺖ ﺑﲔ ‪ %28.9-24.6‬ﻟﻠﻌﻴﻨﺔ ﺑﻒ‪ .‬ﳏﺘﻮﻯ ﺍﻟﻌﻴﻨﺎﺕ ﻣﻦ ﺍﻟﺮﻣﺎﺩ ﺗﺮﺍﻭﺡ ﺑﲔ ‪ %4.3-3.2‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ‬
‫ﺍﻟﻐﺰﺍﻝ ﻭﺗﺮﺍﻭﺡ ﺑﲔ ‪ %4.3-3.5‬ﻟﻠﻌﻴﻨﺔ ﺑﻒ‪ .‬ﻭﺗﺮﺍﻭﺡ ﳏﺘﻮﻯ ﺍﻷﻟﻴﺎﻑ ﺑﲔ ‪ %3.2-2.5‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﻭ‪ %3.1-2.2‬ﻟﻠﻌﻴﻨﺔ ﺑﻒ‪ .‬ﳏﺘﻮﻯ ﺍﻟﺰﻳﺖ ﺗﺮﺍﻭﺡ ﺑﲔ ‪ %1.6-1.5‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭ ‪ %1.7-1.6‬ﻟﻠﻌﻴﻨﺔ‬
‫ﺑﻒ‪ .‬ﻛﻞ ﺍﳌﻌﺎﻣﻼﺕ ﻛﺎﻧﺖ ﺫﺍﺕ ﺗﺄﺛﲑ ﻗﻠﻴﻞ ﻋﻠﻰ ﳏﺘﻮﻯ ﺍﻟﺪﻫﻦ ﺑﻴﻨﻤﺎ ﻛﺎﻧﺖ ﻫﻨﺎﻙ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﰲ ﳏﺘﻮﻯ‬
‫ﺍﻟﱪﻭﺗﲔ ﻭﺍﻟﺮﻣﺎﺩ ﻭﺍﻷﻟﻴﺎﻑ ﻟﻠﻌﻴﻨﺘﲔ‪.‬‬
‫ﰎ ﲡﺰﺋﺔ ﺑﺮﻭﺗﲔ ﺍﻟﻠﻮﺑﻴﺎ ﺍﳋﺎﻡ ﻭﺍﻟﱵ ﰎ ﺗﺰﺭﻳﻌﻬﺎ ﻭﺃﻇﻬﺮ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ‬
‫)‪ (0.05‬ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %70.2 ، %71.4 ، %76.4 ، %80.6 ، %87.5 :‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﺍﳋﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﰲ ﺍﻟﻴﻮﻡ ﺍﻷﻭﻝ ﻭﰲ ﺍﻟﻴﻮﻡ ﺍﻟﺜﺎﱐ ﻭﰲ ﺍﻟﻴﻮﻡ ﺍﻟﺜﺎﻟﺚ ﻭﰲ ﺍﻟﻴﻮﻡ ﺍﻟﺮﺍﺑﻊ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻭﺑﺎﳌﺜﻞ ﻛﺎﻥ ﻫﻨﺎﻙ‬
‫ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ )‪ (0.05‬ﰲ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻵﰐ‪، %75.6 ، %78.3 ، %81.3 ، %89.8 :‬‬
‫‪ %72.6‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻫﺬﺍ ﺑﺎﻟﻨﺴﺒﺔ ﻟﻠﱪﻭﺗﲔ ﺍﻷﻛﱪ ﻭﻫﻮ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ‪.‬‬
‫ﻭﻛﺬﻟﻚ ﻧﺴﺒﺔ ﺍﻷﻟﺒﻴﻮﻣﲔ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ )‪ (0.05‬ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪، %4.0 :‬‬
‫‪ %2.5 ، %1.9 ، %2.3 ، %3.8‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﺑﺎﳌﺜﻞ ﻛﺎﻥ ﻫﻨﺎﻙ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ‬
‫)‪ (0.05‬ﰲ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻵﰐ‪ %1.2 ، %2.2 ، %2.2 ، %1.5 ، %3.6 :‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
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‫ﻭﻛﺎﻧﺖ ﻧﺘﺎﺋﺞ ﺍﻟﱪﻭﻻﻣﲔ ﰲ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻛﺎﻵﰐ‪، %1.6 ، %4.0 ، %3.3 ، %4.0 :‬‬
‫‪ %4.6‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﻧﺘﺎﺋﺠﻪ ﻛﺎﻵﰐ‪ %1.4 ، %3.9 ، %2.8 ، %2.7 ، %4.5 :‬ﻋﻠﻰ‬
‫ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻭﻗﺪ ﺃﻇﻬﺮ ﺍﻟﱪﻭﺗﲔ ﺝ‪-1‬ﺟﻠﻮﺗﻠﲔ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ )‪ (0.05‬ﻭﻛﺎﻧﺖ ﻧﺘﺎﺋﺠﻪ ﰲ ﺍﻟﻌﻴﻨﺔ ﻋﲔ‬
‫ﺍﻟﻐﺰﺍﻝ ﻛﺎﻵﰐ‪ %1.7 ، %0.8 ، %1.3 ، %2.1 ، %2.4 :‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ‬
‫ﻛﺎﻵﰐ‪ %2.1 ، %1.3 ، %1.2 ، %2.3 :‬ﻭ ‪ %1.3‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻭﻛﺎﻧﺖ ﻧﺘﺎﺋﺞ ﺍﻟﱪﻭﺗﲔ ﺝ‪ -2‬ﺟﻠﻮﺗﻠﲔ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻛﺎﻵﰐ‪، %1.9 ، %2.1 ، %2.4 :‬‬
‫‪ %1.7 ، %1.4‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪، %5.1 ، %6.4 ، %2.5 :‬‬
‫‪ %3.9‬ﻭ ‪ %4.5‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﺍﻟﱪﻭﺗﲔ ﺝ‪-3‬ﺟﻠﻮﺗﻠﲔ ﺃﻇﻬﺮ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ )‪ (0.05‬ﻭﻛﺎﻧﺖ ﻧﺘﺎﺋﺠﻪ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﻛﺎﻵﰐ‪ %13.5 ، %12.0 ، %9.8 ، %4.8 :‬ﻭ ‪ %10.4‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ‬
‫ﻛﺎﻵﰐ‪ %11.1 ، %12.5 ، %10.0 ، %4.5 :‬ﻭ‪ %11.7‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻛﻤﺎ ﺃﻇﻬﺮﺕ ﻧﺘﺎﺋﺞ ﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﰲ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %9.2 ، %2.1 ، %2.5 ، %1.2 :‬ﻭ ‪ %6.0‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ‬
‫ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %3.6 ، %1.9 ، %1.7 ، %1.2 :‬ﻭ‪ %6.1‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻛﺎﻥ ﻫﻨﺎﻙ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻟﻠﺠﻠﻮﺑﻴﻮﻟﲔ ﻧﺘﻴﺠﺔ ﻟﻄﺒﺦ ﺍﻟﻌﻴﻨﺎﺕ ﻏﲑ ﺍﳌﺰﺭﻋﺔ‬
‫ﻭﺍﳌﺰﺭﻋﺔ ﰲ ﺍﻟﻴﻮﻡ ﺍﻷﻭﻝ ‪ ،‬ﺍﻟﺜﺎﱐ ‪ ،‬ﺍﻟﺜﺎﻟﺚ ﻭﺍﻟﺮﺍﺑﻊ ﻛﺎﻵﰐ‪ %18.0 ، %18.1 ، %19.0 ، %16.4 :‬ﻭ‬
‫‪ %16.8‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪، %19.9 ، %15.7 :‬‬
‫‪ %22.6 ، %22.5‬ﻭ‪ %23.0‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻛﺬﻟﻚ ﻛﺎﻥ ﻫﻨﺎﻟﻚ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪(0.05‬‬
‫ﻟﻸﻟﺒﻴﻮﻣﲔ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %1.8 ، %3.1 ، %2.8 ، %1.9 :‬ﻭ ‪ %1.4‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ‬
‫ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﰲ ﺍﻟﻴﻮﻡ ﺍﻷﻭﻝ ‪ ،‬ﺍﻟﺜﺎﱐ ‪ ،‬ﺍﻟﺜﺎﻟﺚ ﻭﺍﻟﺮﺍﺑﻊ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
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‫‪ %2.1 ، %1.6 ، %1.6 ، %2.2‬ﻭ‪ %1.3‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻛﻤﺎ ﺃﻇﻬﺮ ﺍﻟﱪﻭﻻﻣﲔ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﻧﺘﻴﺠﺔ‬
‫ﻟﻠﻄﺒﺦ ﻟﻠﻌﻴﻨﺎﺕ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ ‪%5.1 ، %6.3 ، %4.2‬‬
‫ﻭ‪ %5.0‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﻫﻨﺎﻟﻚ ﺯﻳﺎﺩﺓ ﰲ ﺍﻟﻌﻴﻨﺔ ﺍﻟﻐﲑ ﻣﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﰲ ﺍﻟﻴﻮﻡ ﺍﻷﻭﻝ‬
‫ﻛﺎﻵﰐ ‪ %5.1 ، %4.6‬ﰒ ﺗﻨﺎﻗﺺ ﰲ ﺍﻟﺜﺎﱐ ﻭﺍﻟﺜﺎﻟﺚ ﻭﺍﻟﺮﺍﺑﻊ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪، %2.7 ، %2.7 :‬‬
‫‪ %1.8‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﺃﻇﻬﺮ ﺗﻨﺎﻗﺾ ﻣﻌﻨﻮﻱ ﰲ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﺍﻟﻐﲑ ﻣﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﰲ ﺍﻟﻴﻮﻡ‬
‫ﺍﻷﻭﻝ ‪ ،‬ﺍﻟﺜﺎﱐ ‪ ،‬ﻭﺍﻟﺜﺎﻟﺚ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %1.3 ، %1.6 ، %1.4 ، %2.0 :‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﰒ ﺃﻇﻬﺮ‬
‫ﺯﻳﺎﺩﺓ ﰲ ﺍﻟﻴﻮﻡ ﺍﻟﺮﺍﺑﻊ ﺇﱃ ‪ .%2.6‬ﻭﻛﺬﻟﻚ ﰲ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﺃﻇﻬﺮ ﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ‬
‫ﻛﺎﻵﰐ‪ %1.6 ، %1.1 ، %1.2 ، %2.7 :‬ﻭ ‪ %1.0‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﺃﻇﻬﺮ ﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﺍﻟﻐﲑ ﻣﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﰲ ﺍﻟﻴﻮﻡ ﺍﻷﻭﻝ ‪،‬‬
‫ﺍﻟﺜﺎﱐ ‪ ،‬ﺍﻟﺜﺎﻟﺚ ﻭﺍﻟﺮﺍﺑﻊ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %4.0 ، %4.4 ، %4.4 ، %3.4 :‬ﻭ ‪ %5.7‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻭﺑﺎﳌﺜﻞ ﰲ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %7.9 ، %7.5 ، %7.5 ، %5.1 :‬ﻭ‪ %8.8‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﺃﻇﻬﺮ ﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪، %63.8 :‬‬
‫‪ %61.6 ، %62.0 ، %61.0‬ﻭ ‪ %57.0‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﰲ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪ %61.0 ، %62.4 ، %59.5 ، %69.3‬ﻭ‪ %62.1‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﻛﺬﻟﻚ ﺃﻇﻬﺮﺕ ﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ‬
‫ﺍﻟﺬﺍﺋﺒﺔ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪، %8.8 ، %8.0 ، %5.5 :‬‬
‫‪ %9.6‬ﻭ‪ %10.2‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ﻭﺑﺎﳌﺜﻞ ﻛﺎﻧﺖ ﻧﺘﺎﺋﺞ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻵﰐ‪، %6.5 ، %6.6 ، %5.5 :‬‬
‫‪ %6.1‬ﻭ‪ %6.5‬ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻃﺒﺦ ﺭﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 15‬ﺭﻃﻞ‪/‬ﺑﻮﺻﺔ‪ 2‬ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ﺃﻇﻬﺮ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﺗﻨﺎﻗﺼﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻣﻦ ‪ %87.5‬ﺇﱃ ‪120º %29.2‬‬
‫ﻭﺍﻷﻟﺒﻴﻮﻣﲔ ﻣﻦ ‪ %4.0‬ﺇﱃ ‪ %1.2‬ﺑﻴﻨﻤﺎ ﺯﺍﺩ ﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.3‬ﺇﱃ ‪ %4.5‬ﻭﺗﻨﺎﻗﺺ ﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ‬
‫‪ %2.3‬ﺇﱃ ‪ %1.2‬ﻭﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.4‬ﺇﱃ ‪ %1.2‬ﻭﺯﺍﺩ ﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %4.8‬ﺇﱃ ‪%59.5‬‬
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‫ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ ‪ %1.2‬ﺇﱃ ‪ %8.0‬ﻭﻋﻨﺪ ﻃﺒﺦ ﻧﻔﺲ ﺍﻟﻌﻴﻨﺔ ﻃﺒﺦ ﺭﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 20‬ﺭﻃﻞ‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ﺇﳔﻔﺾ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﻣﻦ ‪ %87.5‬ﺇﱃ ‪º %27.5‬ﻟﻠﺒﻮﺻﺔ ﺍﳌﺮﺑﻌﺔ ﰲ ﺩﺭﺍﺟﺔ ﺣﺮﺍﺭﺓ ‪150‬‬
‫ﻭﺍﻷﻟﺒﻴﻮﻣﲔ ﻣﻦ ‪ %4.0‬ﺇﱃ ‪ %1.2‬ﻭﺯﺍﺩ ﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.3‬ﺇﱃ ‪ %4.8‬ﻭﺗﻨﺎﻗﺺ ﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ‬
‫‪ %2.3‬ﺇﱃ ‪ %2.0‬ﻭﺗﺰﺍﻳﺪ ﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.4‬ﺇﱃ ‪ %4.0‬ﻭﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %4.8‬ﺇﱃ ‪%60.2‬‬
‫ﻡ ﳌﺪﺓ ‪º‬ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ ‪ %1.2‬ﺇﱃ ‪ %6.0‬ﻭﻋﻨﺪ ﲢﻤﻴﺺ ﻧﻔﺲ ﺍﻟﻌﻴﻨﺔ )ﺍﻟﻄﺒﺦ ﺍﳉﺎﻑ( ﻋﻨﺪ ‪90‬‬
‫‪ 60‬ﺩﻗﻴﻘﺔ ﺗﻨﺎﻗﺺ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﺗﻨﺎﻗﺼﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻣﻦ ‪ %87.5‬ﺇﱃ ‪ %85.0‬ﻭﺃﻇﻬﺮ‬
‫ﺍﻷﻟﺒﻴﻮﻣﲔ ﺛﺒﺎﺗﹰﺎ ﻭﺗﻨﺎﻗﺺ ﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.3‬ﺇﱃ ‪ %3.6‬ﻭﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.3‬ﺇﱃ ‪ %2.0‬ﻭﺝ‪-2‬‬
‫ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.4‬ﺇﱃ ‪ .%1.2‬ﻭﺗﺰﺍﻳﺪ ﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %4.8‬ﺇﱃ ‪ %6.0‬ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ‬
‫ﻡ ﳌﺪﺓ ‪ 60‬ﺩﻗﻴﻘﺔ ﺃﻇﻬﺮ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ‪ %1.2º‬ﺇﱃ ‪ .%3.0‬ﻭﻋﻨﺪ ﲢﻤﻴﺺ ﻧﻔﺲ ﺍﻟﻌﻴﻨﺔ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪120‬‬
‫ﺗﻨﺎﻗﺼﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻣﻦ ‪ %87.5‬ﺇﱃ ‪ %41.0‬ﻭﺍﻷﻟﺒﻴﻮﻣﲔ ﻣﻦ ‪ %4.0‬ﺇﱃ ‪%1.2‬‬
‫ﻭﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.3‬ﺇﱃ ‪ %4.2‬ﻭﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.3‬ﺇﱃ ‪ %1.2‬ﻭﺗﺰﺍﻳﺪ ﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ‬
‫‪%2.4‬ﺇﱃ ‪ %3.2‬ﻭﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %4.8‬ﺇﱃ ‪ %48.0‬ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ ‪ %1.2‬ﺇﱃ‬
‫‪.%6.0‬‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ﻟﻠﻌﻴﻨﺔ ‪º‬ﻭﻋﻨﺪ ﺍﻟﻄﺒﺦ ﺍﻟﺮﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 15‬ﺭﻃﻞ‪/‬ﺑﻮﺻﺔ‪ 2‬ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪120‬‬
‫ﺑﻒ ﺃﻇﻬﺮ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﺗﻨﺎﻗﺼﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻣﻦ ‪ %89.8‬ﺇﱃ ‪ %32.0‬ﻭﺍﻷﻟﺒﻴﻮﻣﲔ ﻣﻦ‬
‫‪ %3.6‬ﺇﱃ ‪ %1.0‬ﻭﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.5‬ﺇﱃ ‪ %3.0‬ﻭﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.3‬ﺇﱃ ‪ %2.0‬ﻭﺗﺰﺍﻳﺪ‬
‫ﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.5‬ﺇﱃ ‪ %3.0‬ﻭﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %4.5‬ﺇﱃ ‪ %55.0‬ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ‬
‫‪ %1.2‬ﺇﱃ ‪ %6.0‬ﻭﻋﻨﺪ ﻃﺒﺦ ﻧﻔﺲ ﺍﻟﻌﻴﻨﺔ ﻃﺒﺦ ﺭﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 20‬ﺭﻃﻞ ﻟﻠﺒﻮﺻﺔ ﺍﳌﺮﺑﻌﺔ ﰲ ﺩﺭﺍﺟﺔ ﺣﺮﺍﺭﺓ‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ﺗﻨﺎﻗﺺ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﺗﻨﺎﻗﺼﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻣﻦ ‪ %89.8‬ﺇﱃ ‪150º %36.0‬‬
‫ﻭﺍﻷﻟﺒﻴﻮﻣﲔ ﻣﻦ ‪ %3.6‬ﺇﱃ ‪ %1.0‬ﻭﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.5‬ﺇﱃ ‪ %4.0‬ﻭﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.3‬ﺇﱃ‬
‫‪ %1.2‬ﻭﺗﺰﺍﻳﺪ ﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.5‬ﺇﱃ ‪ %3.2‬ﻭﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %4.5‬ﺇﱃ ‪ %50.8‬ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ‬
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‫ﻡ ﳌﺪﺓ ‪ 60‬ﺩﻗﻴﻘﺔ ﺃﻇﻬﺮ ‪º‬ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ ‪ %1.2‬ﺇﱃ ‪ .%7.0‬ﻭﻋﻨﺪ ﲢﻤﻴﺺ ﻧﻔﺲ ﺍﻟﻌﻴﻨﺔ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪90‬‬
‫ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﺗﻨﺎﻗﺼﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﻣﻦ ‪ %89.8‬ﺇﱃ ‪ %88.0‬ﻭﺃﻇﻬﺮ ﺍﻷﻟﺒﻴﻮﻣﲔ ﺛﺒﺎﺗﹰﺎ ﻭﺗﻨﺎﻗﺺ‬
‫ﺍﻟﱪﻭﻻﻣﲔ ﻣﻦ ‪ %4.0‬ﺇﱃ ‪ %4.5‬ﻭﺝ‪-1‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪ %2.3‬ﺇﱃ ‪ %2.2‬ﻭﺗﺰﺍﻳﺪ ﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﻣﻦ ‪%2.5‬‬
‫ﺇﱃ ‪ .%2.8‬ﻭﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﺃﻇﻬﺮ ﺛﺒﺎﺗﹰﺎ ﻭﺯﺍﺩﺕ ﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﺍﻟﺬﺍﺋﺒﺔ ﻣﻦ ‪ %1.2‬ﺇﱃ ‪ .%1.4‬ﻋﻨﺪ ﲢﻤﻴﺺ ﺍﻟﻌﻴﻨﺔ‬
‫ﻡ ﳌﺪﺓ ‪ 60‬ﺩﻗﻴﻘﺔ ﺃﻇﻬﺮﺕ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺍﳉﻠﻮﺑﻴﻮﻟﲔ ﻣﻦ ‪º‬ﺑﻒ ﻏﲑ ﺍﳌﻌﺎﻣﻠﺔ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪120‬‬
‫‪ %89.8‬ﺇﱃ ‪ %45.5‬ﻭﺍﻷﻟﺒﻴﻮﻣﲔ ﻣﻦ ‪ %3.6‬ﺇﱃ ‪ %1.0‬ﻭﺍﻟﱪﻭﻻﻣﲔ ﺯﺍﺩ ﻣﻦ ‪ %4.5‬ﺇﱃ ‪ %6.0‬ﻭﺝ‪-1‬‬
‫ﺟﻠﻮﺗﻴﻠﲔ ﻧﻘﺺ ﻣﻦ ‪ %2.3‬ﺇﱃ ‪ %2.0‬ﻭﺝ‪-2‬ﺟﻠﻮﺗﻴﻠﲔ ﺯﺍﺩ ﻣﻦ ‪%2.5‬ﺇﱃ ‪ %3.0‬ﻭﺝ‪-3‬ﺟﻠﻮﺗﻴﻠﲔ ﺯﺍﺩ ﻣﻦ‬
‫‪ %4.5‬ﺇﱃ ‪ %42.0‬ﻭﺍﻟﱪﻭﺗﻴﻨﺎﺕ ﻏﲑ ﺍﻟﺬﺍﺋﺒﺔ ﺯﺍﺩﺕ ﻣﻦ ‪ %1.2‬ﺇﱃ ‪.%6.0‬‬
‫ﰎ ﺇﺧﺘﺒﺎﺭ ﺍﻟﻌﻴﻨﺎﺕ ﺍﳌﻌﺎﻣﻠﺔ ﻣﻘﺎﺑﻞ ﺍﻷﺧﺮﻯ ﻏﲑ ﺍﳌﻌﺎﻣﻠﺔ ﳊﻤﺾ ﺍﻟﻔﺎﻳﺘﻚ ﻭﺍﻟﺘﺎﻧﲔ ﻭﻣﺜﺒﻂ ﺇﻧﺰﱘ ﺍﻟﺘﺮﺑﺴﲔ‪.‬‬
‫ﻭﻗﺪ ﻭﺟﺪ ﺇﳔﻔﺎﺽ ﻣﻌﻨﻮﻱ )ﰲ ﺣﺪﻭﺩ ﺛﻘﺔ ‪ (0.05‬ﰲ ﲪﺾ ﺍﻟﻔﺎﺗﻴﻚ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪310.3‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻋﻠﻰ ﺃﺳﺎﺱ ﺍﻟﻮﺯﻥ ﺍﳉﺎﻑ‪386.1 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪248.9 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪،‬‬
‫‪201.7‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻭ ‪139.8‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﻌﺎﻣﻠﺔ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ‪،‬‬
‫ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪376.3‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪346.2 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪301.0 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪225.7 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻭ‬
‫‪180.7‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﻌﺎﻣﻠﺔ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ‬
‫ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻭﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺎﺕ ﺍﳌﺰﺭﻋﺔ ﻭﻏﲑ ﺍﳌﺰﺭﻋﺔ ﺃﻇﻬﺮ ﲪﺾ ﺍﻟﻔﺎﻳﺘﻚ ﺗﻨﺎﻗﺺ ﻣﻌﻨﻮﻱ ﰲ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ‬
‫ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪290.3 :‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪268.9 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪229.0 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪،‬‬
‫‪181.5‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻭ ‪128.6‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ‪ ،‬ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ‬
‫‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪353.7‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪311.6 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪270.9 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ‪205.4 ،‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻭ‬
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‫‪162.6‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﻌﺎﻣﻠﺔ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ‪ ،‬ﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ‬
‫ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻭﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻃﺒﺦ ﺭﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 20‬ﺭﻃﻞ ﻟﻠﺒﻮﺻﺔ ﺍﳌﺮﺑﻌﺔ ﰲ ﺩﺭﺍﺟﺔ‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ﺇﳔﻔﺾ ﲪﺾ ﺍﻟﻔﺎﻳﺘﻚ ﻣﻦ ‪310.3‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﺇﱃ ‪º 300.0‬ﺣﺮﺍﺭﺓ ‪150‬‬
‫ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﺇﳔﻔﺾ ﲪﺾ ﺍﻟﻔﺎﻳﺘﻚ ﻣﻦ ‪ 376.3‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﺇﱃ‬
‫ﻡ ﳌﺪﺓ ‪ 60‬ﺩﻗﻴﻘﺔ ﺍﳔﻔﺾ ‪ 300.0º‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ‪ .‬ﻭﻋﻨﺪ ﲢﻤﻴﺺ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪120‬‬
‫ﲪﺾ ﺍﻟﻔﺎﻳﺘﻚ ﻣﻦ ‪ 310.3‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﺇﱃ ‪ 301.0‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﺇﳔﻔﺾ ﲪﺾ‬
‫ﺍﻟﻔﺎﻳﺘﻚ ﻣﻦ ‪ 376.3‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ ﺇﱃ ‪ 352.0‬ﻣﻠﺠﻢ‪100/‬ﺟﻢ‪ .‬ﻭﺃﻇﻬﺮﺕ ﺍﻟﺘﺎﻧﻴﻨﺎﺕ ﺍﳔﻔﺎﺿﹰﺎ ﻣﻌﻨﻮﻳﹰﺎ ﻋﻨﺪ‬
‫ﺗﺰﺭﻳﻊ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ 0.48 :‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.36 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪0.30 ،‬‬
‫ﺟﻢ‪100/‬ﺟﻢ ‪ 0.24 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.20 ،‬ﺟﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ‬
‫ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺍﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‬
‫ﻛﺎﻵﰐ‪ 0.50 :‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.42 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.36 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.30 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪،‬‬
‫‪ 0.22‬ﺟﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ‬
‫ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ‪ .‬ﻭﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻃﺒﺦ ﺭﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 20‬ﺭﻃﻞ ﻟﻠﺒﻮﺻﺔ ﺍﳌﺮﺑﻌﺔ ﰲ‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ﺇﳔﻔﻀﺖ ﺍﻟﺘﺎﻧﻴﻨﺎﺕ ﻣﻦ ‪ 0.48‬ﺟﻢ‪100/‬ﺟﻢ ﺇﱃ ‪ 0.38‬ﺟﻢ‪100/‬ﺟﻢ ‪º‬ﺩﺭﺍﺟﺔ ﺣﺮﺍﺭﺓ ‪150‬‬
‫ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﺇﳔﻔﻀﺖ ﺍﻟﺘﺎﻧﻴﻨﺎﺕ ﻣﻦ ‪ 0.50‬ﺟﻢ‪100/‬ﺟﻢ ﺇﱃ ‪ 0.40‬ﺟﻢ‪100/‬ﺟﻢ‪ .‬ﻭﻋﻨﺪ ﲢﻤﻴﺺ‬
‫ﻡ ﳌﺪﺓ ‪ 60‬ﺩﻗﻴﻘﺔ ﺍﳔﻔﻀﺖ ﺍﻟﺘﺎﻧﻴﻨﺎﺕ ﻣﻦ ‪ 0.48‬ﺟﻢ‪100/‬ﺟﻢ ﺇﱃ ‪º‬ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪120‬‬
‫‪ 0.36‬ﺟﻢ‪100/‬ﺟﻢ ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﺇﳔﻔﻀﺖ ﺍﻟﺘﺎﻧﻴﻨﺎﺕ ﻣﻦ ‪ 0.50‬ﺟﻢ‪100/‬ﺟﻢ ﺇﱃ ‪0.42‬‬
‫ﺟﻢ‪100/‬ﺟﻢ‪ .‬ﻭﺃﻇﻬﺮﺕ ﺍﻟﺘﺎﻧﻴﻨﺎﺕ ﺗﻨﺎﻗﺺ ﻣﺘﺰﺍﻳﺪ ﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺔ ﺑﻒ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ‪ ،‬ﻳﻮﻣﲔ‬
‫‪ ،‬ﺛﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺃﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ 0.26 :‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.18 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪0.15 ،‬‬
‫ﺟﻢ‪100/‬ﺟﻢ ‪ 0.12 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.10 ،‬ﺟﻢ‪100/‬ﺟﻢ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ‬
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‫ﻛﺎﻵﰐ‪ 0.30 :‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.22 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.20 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪ 0.18 ،‬ﺟﻢ‪100/‬ﺟﻢ ‪،‬‬
‫‪ 0.11‬ﺟﻢ‪100/‬ﺟﻢ ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ‬
‫ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻛﻤﺎ ﺃﻇﻬﺮ ﻧﺸﺎﻁ ﺇﻧﺰﱘ ﺍﻟﺘﺮﺑﺴﲔ ﺇﳔﻔﺎﺽ ﻣﻌﻨﻮﻱ ﺑﺎﻟﺘﺰﺭﻳﻊ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪ 22.0‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 11.8 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 10.6 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 8.0 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 8.0 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ‬
‫ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ‪ 25.0‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 12.5 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 10.0 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪9.6 ،‬‬
‫ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ‪ 9.0 ،‬ﻭﺣﺪﺓ‪/‬ﻣﻠﺠﻢ ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ‬
‫ﻼ ﺑﺎﳌﻌﺎﻣﻼﺕ ﺍﳊﺮﺍﺭﻳﺔ ﺍﳌﺨﺘﻠﻔﺔ‪.‬‬
‫ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﰎ ﺗﻜﺴﲑ ﻣﺜﺒﻂ ﺇﻧﺰﱘ ﺍﻟﺘﺮﺑﺴﲔ ﺗﻜﺴﲑﹰﺍ ﻛﺎﻣ ﹰ‬
‫ﻭﻗﺪ ﰎ ﺗﻘﺪﻳﺮ ﺩﺭﺟﺔ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ ﻣﻌﻤﻠﻴﹰﺎ ﻟﻠﻌﻴﻨﺎﺕ ﺍﳌﺰﺭﻋﺔ ﻭﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪ %80.4 ، %77.9 ، %75.3 ، %73.4‬ﻭ ‪ %84.4‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ‬
‫ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪:‬‬
‫‪ %82.4 ، %79.3 ، %75.7 ، %74.2‬ﻭ‪ %83.6‬ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ‬
‫ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﻋﻨﺪ ﻃﺒﺦ ﺍﻟﻌﻴﻨﺎﺕ ﺍﳌﺰﺭﻋﺔ ﺃﻇﻬﺮﺕ ﻗﻴﻤﺔ ﻫﻀﻢ‬
‫ﺍﻟﱪﻭﺗﲔ ﺯﻳﺎﺩﺓ ﻣﻌﻨﻮﻳﺔ ﻭﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %88.8 ، %87.5 ، %87.2 ، %86.2 :‬ﻭ‪%88.5‬‬
‫ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ‬
‫ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﻛﺎﻧﺖ ﺍﻟﻨﺘﺎﺋﺞ ﻛﺎﻵﰐ‪ %87.9 ، %86.6 ، %86.3 ، %85.4 :‬ﻭ‪%88.3‬‬
‫ﻟﻠﻌﻴﻨﺔ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻡ ﻭﺍﺣﺪ ﻭﺍﳌﺰﺭﻋﺔ ﻟﻴﻮﻣﲔ ﻭﺍﳌﺰﺭﻋﺔ ﻟﺜﻼﺛﺔ ﺃﻳﺎﻡ ﻭﺍﳌﺰﺭﻋﺔ ﻷﺭﺑﻌﺔ ﺃﻳﺎﻡ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫ﻡ ﳌﺪﺓ ‪ 30‬ﺩﻗﻴﻘﺔ ‪º‬ﻋﻨﺪ ﺍﻟﻄﺒﺦ ﺍﻟﺮﻃﺐ ﲢﺖ ﺿﻐﻂ ‪ 20‬ﺭﻃﻞ ﻟﻠﺒﻮﺻﺔ ﺍﳌﺮﺑﻌﺔ ﰲ ﺩﺭﺍﺟﺔ ﺣﺮﺍﺭﺓ ‪150‬‬
‫ﺯﺍﺩﺕ ﻗﻴﻤﺔ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻏﲑ ﺍﳌﺰﺭﻋﺔ ﻣﻦ ‪ %73.4‬ﺇﱃ ‪ %86.0‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﺯﺍﺩﺕ‬
‫ﻡ ﳌﺪﺓ ‪ 60‬ﺩﻗﻴﻘﺔ ﺯﺍﺩﺕ ‪º‬ﻣﻦ ‪ %74.0‬ﺇﱃ ‪ .%87.0‬ﻭﻋﻨﺪ ﲢﻤﻴﺺ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪120‬‬
‫‪26‬‬
‫ﻗﻴﻤﺔ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ ﻣﻦ ‪ %73.4‬ﺇﱃ ‪ %84.0‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﺯﺍﺩﺕ ﻗﻴﻤﺔ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ ﻣﻦ ‪ %74.2‬ﺇﱃ‬
‫‪.%84.0‬‬
‫ﻭﻗﺪ ﲤﺖ ﺩﺭﺍﺳﺔ ﺗﺄﺛﲑ ﺍﻟﺴﺒﺴﺘﺮﺍﺕ ﻋﻠﻰ ﻗﻴﻤﺔ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ ﺑﺎﺳﺘﺨﺪﺍﻡ ﻋﻴﻨﺎﺕ ﳐﻠﻮﻃﺔ ﻣﻦ ﺍﻟﻌﻴﻨﺔ ﺍﳌﻌﺎﻣﻠﺔ‬
‫ﺣﺮﺍﺭﻳﹰﺎ ﻣﻊ ﺍﻟﻌﻴﻨﺔ ﻏﲑ ﺍﳌﻌﺎﻣﻠﺔ ﻭﻛﺎﻧﺖ ﺍﳋﻠﻄﺔ ﰲ ﺍﻟﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻋﻠﻰ ﺍﻟﻨﺤﻮ ﺍﻷﰐ‪0.192 :‬ﺟﻢ ‪0.096 ،‬‬
‫ﺟﻢ ‪ 0.048 ،‬ﺟﻢ ‪ 0.00 ،‬ﺟﻢ ‪ 0.288 ،‬ﺟﻢ ‪ 0.336‬ﺟﻢ ﻭ‪ 0.384‬ﺟﻢ ﻣﻦ ﺍﻟﻌﻴﻨﺔ ﺍﳋﺎﻡ ﻣﻊ ‪0.192‬‬
‫ﺟﻢ ‪ 0.288 ،‬ﺟﻢ ‪ 0.336 ،‬ﺟﻢ ‪ 0.384 ،‬ﺟﻢ ‪ 0.096 ،‬ﺟﻢ ‪ 0.048 ،‬ﺟﻢ ﻭ ‪ 0.00‬ﺟﻢ ﻣﻦ‬
‫ﻡ ‪ º‬ﻡ ﻭ ‪º120‬ﺍﻟﻌﻴﻨﺎﺕ ﺍﳌﻌﺎﻣﻠﺔ ﺣﺮﺍﺭﻳﹰﺎ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﻗﺪ ﻛﺎﻧﺖ ﺍﳌﻌﺎﻣﻠﺔ ﺍﳊﺮﺍﺭﻳﺔ ﻟﻠﻌﻴﻨﺎﺕ ﲢﻤﻴﺼﹰﺎ ﰲ ﺩﺭﺟﺔ ‪90‬‬
‫ﻡ ﲢﺖ ﺿﻐﻂ ‪º 10‬ﳌﺪﺓ ‪ 45 ، 30‬ﻭ‪ 60‬ﺩﻗﻴﻘﺔ ﺑﺎﻹﺿﺎﻓﺔ ﻋﻠﻰ ﻃﺒﺦ ﺭﻃﺐ ﰲ ﺩﺭﺟﺔ ﺣﺮﺍﺭﺓ ‪115.5‬‬
‫ﻡ ﲢﺖ ﺿﻐﻂ ‪ 15‬ﺭﻃﻞ‪/‬ﺑﻮﺻﺔ‪ 2‬ﳌﺪﺓ ‪ 45 ، 30‬ﻭ‪ 60‬ﺩﻗﻴﻘﺔ ﻭﻗﺪ ﺗﺮﺍﻭﺣﺖ ﻧﺘﺎﺋﺞ ﻫﻀﻢ ‪º‬ﺭﻃﻞ‪/‬ﺑﻮﺻﺔ‪ 2‬ﻭ ‪120‬‬
‫ﺍﻟﱪﻭﺗﲔ ﺑﲔ‪ %84.2-73.4 ، %84.0-%73.4 :‬ﺑﺎﻟﻨﺴﺒﺔ ﻟﻠﻌﻴﻨﺎﺕ ﺍﶈﻤﺼﺔ ﻛﻤﺎ ﺗﺮﺍﻭﺣﺖ ﰲ ﺍﻟﻄﺒﺦ‬
‫ﺍﻟﺮﻃﺐ ﺑﲔ ‪ %82.0-73.4‬ﻭ ‪-73.4 ، %85.0-73.4 ، %84.0-73.4 ، %83.0-73.4‬‬
‫‪ %85.5‬ﻭ‪ %86.2-73.4‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪ .‬ﻭﺑﺎﳌﺜﻞ ﻟﻠﻌﻴﻨﺔ ﺑﻒ ﺗﺮﺍﻭﺣﺖ ﻗﻴﻤﺔ ﻫﻀﻢ ﺍﻟﱪﻭﺗﲔ‬
‫ﻣﻦ ‪ %84.0-74.2 ، %83.8-74.2 ، %79.8-74.2 ، %84.6-74.2 ، %79.7-74.2‬ﻭ‬
‫‪ %84.6-74.2‬ﺑﺎﻟﻨﺴﺒﺔ ﻟﻠﻌﻴﻨﺎﺕ ﺍﶈﻤﺼﺔ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ ‪ ،‬ﻭﺗﺮﺍﻭﺣﺖ ﰲ ﺍﻟﻄﺒﺦ ﺍﻟﺮﻃﺐ ﻣﺎ ﺑﲔ ‪-74.2‬‬
‫‪-74.2 ، %85.0-74.2 ، %85.0-74.2 ، %84.8-74.2 ، %84.8-74.2 ، %80.2‬‬
‫‪ %85.4‬ﻟﻠﻌﻴﻨﺔ ﻋﲔ ﺍﻟﻐﺰﺍﻝ ﻋﻠﻰ ﺍﻟﺘﻮﺍﱄ‪.‬‬
‫‪27‬‬
CHAPTER ONE
INTRODUCTON
28
1. INTRODUCTON
1.1. Importance of cowpea:
Cowpeas (Vigna unguiculata and Vigna Senensis) are important grain
legumes in East and West Africa as well other developing countries (Dovlo et al.
1976). The beans are also known by other names such as blackeye beans,
southern peas, colossus peas and crowther peas. The pulse is indigenous to Africa
(Okigbo, 1986), though it is now grown in other continents, such as Central
America (Bressani et al. 1961) as well as North and South America and Asia.
Higher meat prices during recent years and the need for protein- rich foods have
led people in the most developing countries to shift their consumption to
cowpeas and other grain legumes. Cowpea is grown mainly for the seed and
sometimes pods in West Africa, India and South America, but it is grown for both
seeds, pods and leaves in East Africa. It is also utilized as fodder and as quick
growing cover-crop. It improves soil fertility because of its ability to fix nitrogen
efficiently (up to 240 kg N per hectare) and can leave a fixed- N deposit in the soil
of up to 60 – 70 kg/ha for the succecding crop (Rechie, 1985; Kachare, et al.
1988). Like other grain legumes, cowpeas are a good source of energy, proteins
(amino acids), vitamins, minerals and deitary fiber. Legumes are sometimes
refserred to as “poor man’s meat” or the “rich man’s vegetable” ( Walker, 1981).
Cowpea is most widely consumed legume in Nigeria as it represents a cheap
source of dietary protein. Of the different species of bean consume in the country,
cowpeas in particular have attracted attention as possible home grown sources of
protein and successive selections have made it possible to introduce several high
yielding varieties with desirable packages of nutrients and with resistance to pest
29
and microbial infections (Ologhobo et al, 1983). Cowpea are major sources of
protein in developing countries where consumption of certain animal foods is
taboo because of religious or customary beliefs. This trend has been obsorved in
parts of northern Nigeria where pork and pork products, horse meat, donkey meat
and meat of camels and asses are avoided by muslims. Cowpeas are a major
source of thiamin and niacin and also contain reasonable amount of other watersoluble vitamins, riboflavin, pyridoxine and folacin. In addition, they supply the
essential minerals, calcium, magnesium, potassium, iron, zinc and phosphorus
(Aykroyed et al. 1982). In Sudan, people relied mostly on other available
leguminous crops (Ahmed and Nour, 1990).
Cowpea proteins provide the three esential amino acids lysine threonine
and methionine, (Dhankher et al 1990. Sosulski, 1987; Fashakin, 1986; Philips,
1987; Saeed, 1977; El Hardallou, 1980). Intense efforts to find alternative sources
of proteins from plants adapted to adverse conditions are being conducted around
the world (Siddhuraju et al. 1995; Bravo et al., 1994; Bhattacharya et al. 1994).
Despite the importance of cowpea (Vigna unguculata L. Walp.) as afood crop in
tropical and subtropical regions, especially in West Africa (Rachie, 1973; R.D
phillips 1982 a), little work has been carried out on the characterization of its
major seed proteins in comparison to other legume species such as Glycine max or
Phaseolus vulgaris. A major globulin protein was identified by Joubert (1957).
and (Carasco et al. 1978) reports further details of protein subunits present in both
mature and developing cowpeas. Sefa–Dedeh and Stany, 1979a,b, reports further
details of the solubility parameters of cowpea proteins.
1.2. Classification of Cowpea:
30
Cowpea belongs to the family leguminosae . It is known as black eye beans or
southern peas, colossus peas and crowpeas . The pulse is indingenous to Africa
though it is now grown in other non African countries (Uzogara, 1992a, b).
31
1.3. Antinutritional factors:
Legumes are generally known to contain various natural constitunets
which affect their nutritional quality. Some of these components are proteins
which inhibit specific enzyme activities for example the inhibitors of protease and
amylases. Orhers are haemagglutinins, saponins, tannins and antivitamins
(Witaker and Feeney, 1973; Liener, 1969). Cowpeas contain antinutritients such
as polyphenols, tripsin inhibitors, lectins and phytates which may decrease protein
digestibility and reduce protein quality (Bressani and Elias, 1978). The
availability of nutrients in plant foods may be affected by natural complexing
agents. This is particularly true in the case of seeds containing phytic acid, such as
legumes, cereals, and oil seeds.
In fact, because of its highly reactive structure, at different pH levels
phytic acid can complex proteins as well as mono- and divalent cations . Phytic
acid and its chemical and nutritional effects have been extensively reviewed
(Cheryan 1980; Reddy et al. 1982). From a nutritional point of view, many studies
have concentrated on the metal ion chelating property of phytic acid (Evdman
1981), its binding of zinc and formation of less soluble complexes that reduce zinc
availability (Prasad, 1979; Morris and Ellis, 1980). The interaction of phytic acid
with proteins has been studied mainly in soy beans. Such studies describe the
formation of phytate protein complexes, their effects on protein solubility and
related properties, and methods for phytic acid removal (Okubo et al., 1975, 1976;
O’ Dell and DeBoland, 1976; De Rham and Jost 1979; Omosaiye and Cheryan
1979; Honing et al. 1984). However, the nature of the phytate- protein interaction
is not completely understood, and its nutritional effects on protein availability still
32
needs clarification. Phytate–protein binding is affected by several factors, such as
the characteristics of the protein matrix (Carnoval E., et al .1987). However, the
acceptability and utilization of legumes as food has been limited due to the
presence of relatively high concentrations of certain antinutritional factors such as
lectins, protease inhibitors, α-amylase inhibitors, allergens, polyphenols, and
phytic acid (Liener, 1994). Phytic acid, the hexaphosphate ester of myoinositol, is
a major phosphorus storage constituent of most cereals, legumes and oilseeds, The
amount of phytic acid is these products varies from 0.5 to 6% and accounts for
between 50 and 90% of the total phosphorus. The ability of phytate to complex
with proteins and with minerals and the consequences of these interactions have
attracted considerable interest from both chemical and nutritional view points
(Kevin et al. 1987). Tannins are high molecular weight polyphenolic compounds
that have the ability to bind with proteins, through hydrogen bonding with
peptides linkages, tannins precipitate proteins from aqueous solutions rendering
plant proteins relatively indigestible and reducing enzyme activity (Van sumere et
al. 1975). Although heat treatment will effectively eliminate most of these
undesirable substances; careful control of processing conditions is essential to
prevent both functional as well as nutritional damage to the protein. On the other
hand, the breeding of varieties or strains of cowpea with low levels of one or more
of the antinutritional factors offers a much more satisfactory long- term solution
to this problem (Ologhobo et al. 1983).
1.4 Processing of cowpeas:
Processing of cowpeas and legumes in general is essential to make them
nutritious, nontoxic, palatable and acceptable. The constraints to maximum
33
utilization of cowpeas can be overcome by appropriate processing technology.
Processing can be classified into domestic and industrial techniques. The domestic
processing techniques that are practiced in villages in developing countries
include dehulling, grinding, soaking, germination, fermintation, addition of salts,
wet and dry heat treatments, cooking and roasting. Theses processes can also be
achieved in the food industry. The industrial processing techniques that are not
common in the villages include canning, roasting, extrusion cooking, formation of
protein concentrates and isolates and texturized vegtable proteins. Processing can
further be divided into primary and secondary processes. Primary processes yield
storable products to be used as and when required and include smoaking,
dehulling, grinding and milling. Secondary processes are involved in the
preparation of final consumer products from cowpeas and include various forms
of heat treatment, such as boiling, steaming, cooking, alkaline treatment, roasting
and deep fat frying (Uzogara et al. 1992).
1.5 Utilization of cowpea:
The potential of cowpeas to contribute more to African or tropical diets
has been investigated (IDRC, 1973) with an emphasis on reducing postharvest
losses (Beucchat, 1983), on developing appropriate technologies to alleviate the
heavy labor imputs required in many traditional preparations (Reichert et al. 1979;
Hudda, 1983). Cowpeas were chosen as the source of nutrients because of their
relatively high protein content and their amino acid profile, which is both
superior to and complementary to that of cereal grains (Dovlo et al. 1976;
Rechie, 1985). Protein- energy malnutrition of infants is one of the major
nutritional problems in the world. It is due to several causes including lack of
34
weaning foods, the preparation of weaning foods with inadequate protein content,
and to the use of foods too low in energy density to satisfy the needs of the
growing infants (Oyus et al., 1985). A weaning food commonly used in Nigeria is
composed largely of sorghum with limiled amount of dried–milk powder, usually
in the ratio 5:1. Such mixtures have been shown to be poor in protein content and
quality (Akinrele and Bassir, 1967; Oyeleke, 1977) Amore suitable weaning food
has been prepared based on soybean and maize flour enriched with vitamins
(Akinrele et al., 1970).
An attempt was made to improve the quality of the popular mixture by
replacing part of the sorghum with cowpeas, Vigna unguiculatea, (Oyus et al.,
1985). Cowpea is being successfully used in child-feeding programme (Ologhobo,
1983). In tropical Africa cowpeas are primarily used in the form of dry seed
cooked as a pulse in large variety of dishes. Preference is for brown, white or
cream seeds with a small eye and wrinkled or rough seed coat. In many areas of
both West and East Africa the tender green leaves are cooked like spinach or as
relish. Green beans or cut green pods are used as avegetable of secondary
importance. Cowpeas are also grown for fodder, ground cover or green manure
but to a much lesser extent than for pulse. Cowpea is practically never grown as a
sole crop in Africa. In Asia the pulse uses of cowpeas are important primarily in
the drier regions such as India where increasing amounts of crop are used in dhal.
In South America cowpeas are grown mainly as pluse. In the USA dry seed is
grown mainly in California and Western Texas, whereas green peas for canning
and freezing (Rachie, 1985).
1.6. Production of Cowpea:
35
Worldwide cowpea production in 1981 was estimated at 2.27 million
tonnes from 7.7 million hectares. Cowpeas are grown extensively in 16 African
countries, with this continent producing two–thirds of the total. Two countries–
Nigeria and Niger- prduce 850,000 t annually or 49.3 per cent of the world crop.
The second–highest producing country is Brazil where 600,000 t of dry seeds or
26.4 percent of the worldwide total was produced in 1981. Other major producers
in Africa include Burkina Faso (95,000t), Ghana (57,000t), Kenya (48,000 t).
Uganda (42,000t ), and Malawi (42,000 t). Tanzania, Senegal and Togo each
produces annually from 20,000 ton to 20.000 ton. Current estimates of production
vary widely according to source, but the statistics are probably conservative for
example, Asian production, including that of long beans as a vegetable, may be
under estimated by a factor of 10 or at a level of about 1 millon hectares,
concentrated in India, Sirlanka, Burma, Bangladesh, Philippines, Indonesia,
Thailand, Pakistan, Nepal, China and Malaysia. India alone is estimated to
cultivate more than half or 500,000 ha for dry seed fodder, green pods and green
manure. Similarly, production estimates may be low for Africa and the Western
Hemisphere where cowpeas are traditionally included as associated crops in
peasant – farming system. Thus, realistic production levels may approach or
exceed 2.5 million tonnes of dry seeds on about 9 million hectares.
The only developed country producing large amounts of cowpeas is USA.
(60.000 ton). Low yields are significant attribute of production estimates,
particularly in Africa and Asia where 240-300 kg/ha are typical (K.O Rachie et
al., 1985). In Sudan the yield is 1070 – 1190 kg /hectare.
Objectives:

The obectives of this study were to assess:
36

The effect of antinutritional factors (tannin, phytic acid , and trypsin
inhibitors) on cowpea protein digestibility.

The effect of dry cooking (roasting), wet cooking (autclaving) on
antinutritional factors as well as protein digestitibity.

The effect of germination on antinutritional factors and protein
digestibility.

The effect of heat tretment on protein fractions and protein
digestibility.
37
CHAPTER TWO
LITERATURE REVIEW
38
2. LITERATURE REVIEW
2.1. Nutritional value:
Cowpea is eaten in the form of dry seeds, green pods and tender green
leaves (Rachie 1985). Like other grain legumes, cowpeas are a good source of
energy (Carbohydrate range was 54.4–63.6 g/100g, fat ether extract range was
1.2–1.4 g /100g). Protein range was 23.1–31.3 g/100g, Vitamins, thiamin range
was 0.77–0.8 mg/100g, riboflavin range was 2.5 –2.0 mg/100g. Niacin range was
3.48–2.8 mg/100g, minerals (ash) range was 3.5-3.0g/100g and deitary fiber
(crude fiber) range was 6.30–0.64 g/100g, amino acids range were lysine 6.6–8.1,
Histidine 2.9–4.47, Arginine 5.4–8.0, Threonine 3.6-45, valine 4.9–57, Isoleucine
4.2–4.8, leucine 7.6–8.5, Tyrosine 2.2–3.6, Phenylalanine 5.5 – 6.2, Methionine
1.5 – 2.3, crude protein 23.1–31.3 (Walker, 1981). They are useful sources of
good quality protein during hungry season. Cowpea contain about 20-30 %
protein. But protein digestibility is low in the beans (Onigbinde et al. 1990). The
food value of cowpeas is highly rated by the nutritionists, as they can provide
supplementary proteins to traditional diets based on cereals, starchy roots and
tubers (Aykroyd et al., 1982; Matthews 1989). Cowpeas are rich in lysine and
other essential amino acids but low in sulphur amino acids (Bressani 1985;
Kochhar et al., 1988; Armu 1990). The high lysine content of cowpea makes it an
excellent improver of the protein quality of foods low in lysine, such as cereals,
which are low in lysine but rich in sulphur amino acids.
The quality of a food legume is highest when such food contains high level
of sulphur amino acids (Bressani et al. 1980).
39
Maximum nutritional benefits are therefore achieved by complementing
cereals with cowpeas in the right amounts so that cereal-cowpea mixes yield
amino acid scores closer to the FAO/WHO/UNU standard found in meats, fish
and egg. Addition of methionine to cowpea protein (Bressani 1985; Sherwood et
al., 1954) significantly increased protein quality estimated by biological value
(BV) and net protein utilization (NPU). Cowpeameal (CM) BV % was 58.17 ±
2.31 and NPU% was 50.6 ± 1.83, CM + cystine BV% was 80.25 ± 1.87 and
NPU% was 72.74 ± 0.94, CM + cystine + methionine BV % was 94.61 ± 1.26 and
NPU % was 82.12 ± 1.07, CM + methionine BV% was 95.84 ± 1.45 and NPU %
was 81.46 ± 0.87, Albumin BV % was 101.72 ± 2.54 and NPU% was 99.52 ±
1.85. Cowpea – cereal mixtures provide the highest quality protein at a weight
ratio of 45 parts cereal to 15 parts cowpea (Bressani 1985).
Cowpeas are a major source of protein, water soluble vitamin and essential
minerals in developing countries (Aykroyd et al., 1982).This is important, since
in most developing countries milk is hardly an important part of the diet; the need
for calcium can be met by consuming cowpeas and other vegetable foods.
The low sodium content of cowpeas makes it a good food for
individuals on low sodium diets, while their high potassium content
should be of special interest to those individuals who take diurectics to
control hypertension and who need increased intake of K+ to replace
that excreted. Cowpeas are also low in fat and contain no cholestrol.
Immature cowpea seeds are good source of vitamin A, beta carotene
and vitamin C (Eheart et al. 1948). Cowpeas also have a great
40
potential in up grading traditional weaning foods based on cereal paps
(Odum et al. 1981; Oyeleke et al. 1985). The beans are sometimes fed
as the main food to infants in developing countries unless such
infants show intolerance to the cowpea diet. A food product for
weaning small children made of 75 % cereal grain and 25% cowpea
would be about 13% good quality protein (Bressani, 1985). Cowpea
are relatively cheap compared to meat foods and, as they have a high
(50 – 65%) carbohydrate content, act as high energy foods for
peasants and nomadic farmers (Longe 1980). Cowpeas also add
variety to monotonous high carbohydrate staples common in the
tropics. Starch contributes about 30 –50% of cowpea carbohydrate and
as in other food legumes over 50% of the starch is in the form of
amylose. Srinavasa– Rao (1976) showed that high amylose content
caused slow digestibility. However, carbohydrate digestibility of
cawpea was increased in vitro by baking, roasting and germination
and these processes might also facilitate in vivo carbohydrate
digestibility (Srinavasa–Rao 1976; Geervani et al., 1981; Reddy et al.
1984; Carbezas et al . 1982). In addition, cowpeas also contain some
indigestible sugars known as oligosaccharides.
41
These oligosaccharides, stachyose, raffinose and verbaseose
cause gas or flatus in some individuals who consume cowpeas
(Uzogara et al . 1992a).
2.2. Protein fractionation
2.2.1. Nitrogen solubility (NS):
Cowpea protein extractability at the isoelectric point 40% of the extractable protein; unlike
most legumes which showed a protein extractability of about 10% or less at the isoelectric
point. This high extraction at the isoelectric pH has been attributed to presence of neutral salts
in the extraction buffers or the composition of the proteins since some albumins and globulins
may not precipitate at the isoelectric pH (4.4) which affected by ionic strength and pH (Sefa –
Dedeh et al. 1979). Solubility is an important property governing the functional behavior of
proteins and their potential application to food processing . Dennaturation implicates damages
to functionality and is usually measured as a loss of solubility. Generally soluble proteins
posses superior functional attributes for moat applications in food processing. Protein
functionality is dependant on hydrophobic, electrostatic, and steric parameters of the proteins,
which are essential for defining the protein structure (Nakai, 1983).
2.2.2. Protein fractions classification:Protein from cowpea seeds can be recovered in sex solubility fractions.
Fraction I contained salt soluble protein globulins, the value for fraction I ranges
from 65.7 to 79.7 %. Fraction II contains water soluble protein albumins which
ranged from 4.0 to 12.3 % Fraction III contained alcohol soluble protein. Prolamin
from 1.4 to 4.0 % . Fraction IV contained G1-glutelins range from (0.9 to 3.0% .
Fraction V. contains G2–glutelins range from 1.4 to 2.9% . Fraction VI contains
G3–glutelin range from 9.0 to 14.0% and insoluble protein rangs from 0.5 to 3.0%
(Nugdallah and El Tinay, 1997).
42
Cowpea proteins classification depends on selective solubility of proteins in
defferent extraction solvents
According to Fruton et al. (1959) proteins are divided into five major groups,
albumins which are water–soluble, globulins which are salt-soluble, prolamins
which are 70–80% ethanol–soluble, glutelins which are sodium hydroxide–
soluble, and sclero–proteins which are insoluble in aqueous solvents.
Osborne (1924) reported that globulins are the major storage proteins of
legumes and they require appreciable salt concentration for solubilization and
they account for 50–70% of the total seed proteins. Other studies on legume
proteins have reached the same conclusion (Romero et al., 1975; Cjakrborty et al.,
1979). Chan-CW. et al (1994), reported that the abundance of cowpea CV.
California Blackeye No.5 seed protein fractions was in the following order:
globulins > albumins > glutelins > prolamins. The globulins contained 4 major
polypeptides with molecular mases of 65, 60, 59 and 50 KDa, 3 of which were
covalently bound with carbohydrate. The albumins cotained 4 major polypeptides
with molecular masses of 99, 91 32 and 30 KDa. The alkali- soluble glutelin
fraction was mainly composed of polypetides in the molecular mass range 44-62
KDa. Polypeptides of molecular mass 105, 62,59 and 54 KDa, were found in the
prolamin fraction. The glutelin and prolamin fractions were high in essential
amino acids compared with the other 2 fractions.
2.2.2.1 Albumins and globulins:
43
Albumins and globulins are very heterogeneous and differed in
polypeptide composition. Extraction with water and then with NaCl to solubilize
what is termed albumins and globulins respectively. These descriptions are
inaccurate since water also extracts the lower molecular weight nitrogen (LMWN)
as well as albumin. In addition, because of endogenous salt in the grain some
globulins are also extracted (Wilson et al., 1981). The globulins constituted 82.6%
of the total seed proteins and the albumins 8.6%. Albumins were richest (3.76
g/100 g protein) in methionine, (Dhankher et al., 1990). The cross contamination
of the water–and salt soluble proteins of legumes attributed mainly to the native
ionic strength of been flours and the low flour–to–solvant ratio. Thus a sharp
distinction between albumins and globulins on the basis of the solubility can not
made (Bhatty, 1982). Water – and saltsoluble proteins together accounted for 63
– 83 % of the total nitrogen of chickpeas, cowpeas and dry beans (Deshpande et
al., 1987) . Six chickpeas were analyzed for their protein fractions, albumin
content ranged from 8.4 to 12.3% and globulin content ranged from 53.4 to 60.3
% (Dhawan et al., 1992).Several genetic lines of field bean seeds were anayzed
for electrophoretic patterns of the globulin and albumin fractions. Globulin
constituted the major fraction of seed protein (Pasqualini et al., 1992). The general
deficiency of lysine in most cereals e.g sorghum and corn is essentially the
consequence of their low content of albumins and globulins (FAO, 1981).
Globulins account for 50 – 75% of the total seed protein (Osborne, 1924).
Nugdallah and El Tinay (1997) reported that Albumins and globulins ranged form
4.0 – 12% and 66 – 80% respectively.
2.2.2.2. Prolamin:
44
Prolamin can be defined as a portion material extracted at room
temperature by aqueous alcohol; free of reductant and salt, from a corn meal
deprived of lipids and salt soluble protein. Prolamin proteins are synthesized in
the developing endosperm where they form protein bodies–within the rough
endoplasmic reticulum because they account for more than half of the total seed
protein (Landry and Moureaux, 1981). Prolamin content ranged from 3.1 to 6.9%
of total seed protein in chickpea seeds (Dhawan et al., 1991). Prolamin content
was 2.6% in cowpea seeds (Dhankher et al. 1990). The ethanol soluble protein
fraction was 0.97% in cowpea seeds (Deshpande and Nielsen, 1987). Nugdallah
and El Tinay (1997) reported that prolamins ranged from 1.4 – 4.0% for cowpea.
2.2.2.3 Glutelin:
Glutelines are defined as including these proteins that are either soluble in
dilute aqueous alkali or insoluble in neutral aqueous solutions, saline solutions or
alcohol (Osborne, 1924). After albumin, globulin and prolamin proteins were
removed from corn, it was found that the addition of 2 – ME to 70% ethanol and
0.5% sodium acetate removed protein from corn (Paulis et al., 1969 ). This protein
was thought be zein like in solubility but later it was termed alcohol – soluble
reduced glutelin based on the definition that all proteins remaining after removel
of salt and alcohol soluble proteins were glutelins (Paulis and Wall, 1971).
Glutelins consists of several different polypeptide chains linked by disulfide
bonds to form an isoluble three–dimensional matrix (Nielsen et al., 1970 ).
Glutelins are associated with lower molecular weight proteins through
noncovalent bonding: they consist mainly of two categories of polypeptide linked
45
by disulfide bonds. Alcohol soluble and alcohol insoluble glutelins are two types
of polypeplide deposited in different subcellular structures. The alcohol soluble
polypeptides resemble prolamins but have significant structural differences
(Paulis, 1982).
Corn grain contains three glutelin subgroups called G1, G2 , and G3 –
glutelin (Landry and Moureaux, 1970). Cowpea cultivars contain three glutelin
fractions called G1, G2, G3 - glutelin (Nugdallah and El Tinay, 1997).
2.2.2.3.1. G1-glutelin:
G1–glutelin (Zein–Like) has an amino acid composition somewhat similar
to zein, but with higher levals of glycine, methionine, histidine and proline and
lower levels of a spartic acid leucine and isoleucine, G1 - glutelin was previously
recoverd as a part of glutelin fraction, zein apears to be cross – linked to glutelin
through disulfide bonds (Paulis et al., 1969). Nugdallah and El Tinay (1997)
reporte that G1 – gutelins ranged from 0.9 – 3.0% for cowpea.
2.2.2.3.2. G2 - glutelin:
G2 – glutelin (glutelin–like ) isolated at pH 10, can be fractionated on the
basis of their extractability at pH 3 (Misra et al. 1972). Acid soluble G2- glutelin
exhibits some general characteristics of cereal prolamin. They are rich in proline,
glutamic acid or glutamine and typified by high histidine and poor lysine and
aspartic acid or asparagine. Moreover, they may be extracted both by acidic and
alcoholic media . Indeed, amino acid composition of acid soluble G2 – glutelins
and water soluble, alcohol soluble–glutelins are nearly identical (Landary and
46
Moureaux, 1981).Consequently, the acid insoluble G2–glutelins are not removed
by alcoholic extraction.
Because of their extractability and amino acid composition, especially
their relatively high lysine cotent, they may be regarded as being similar to G3 –
glutelins. Thus fraction IV contains both acid – soluble and acid – insoluble. G2 –
glutelins, rich in histidine, which might be called prolamine–like and glutelin–
like, respectively. Amount of G2–glutelins isolated at step (5) depends on
conditions used earlier to extract salt – soluble and alcohol – soluble proteins ( L
andry and Moureaux, 1981). Nugdallah and El Tinay (1997) reported that G2 –
glutelins ranged from 1.4 – 2.9% for cowpea.
2.2.2.3.3 G3 - glutelin
There are polypeptide which did not separate clearly on SDS –
polyacrylamide gel (Misra et al., 1972). Moreover, G3–glutelins having an amino
acid composition similar to that of salt soluble proteins (though richer than them
in hydrophobic residues and with lower cysteine content than other glutelin
subgroups). It appears to be non extractable by 2-ME in a saline or alcoholic
medium or by their combination. The in ability of extract G3–glutelins with such
media may be related to noncovalent interpolypeptide bonds (Landry and
momreaux, 1981).
G3–glutelins exist at the earliest stages of grain development before zein
accumulation so they may consist of membrane protein from cell organelles such
as mitochondria or ribosomes (Landry and Moureaux, 1976). This hypothesis
lends support to the existence of noncovalent bonds in glutelins since hydrophobic
47
interactions stabilize these membranes and their multimeric enzymes. This is also
a close Parallel between the decrease in the amount per grain of salt soluble
proteins and the increase of G3–glutelins observed during grain maturation.
Therefore, G3–glutelins include membrane–bond proteins, some naturally
associated salt-soluble proteins and some proteins altered during extraction. As G3
– glutelins could contain membrane proteins, all the more as detergent is
necessary to it dissolution (Landary and moureaux, 1981). Alkali soluble protein
fraction was 11% in legumes (Des hpande and nielsen, 1987). The glutelin
fraction in cowpea seeds was 6.4% of the total seed proteins (Dhankher et al.
1990) . In six chickpeas cultivars analyzed for their protein fractions glutelins
content ranged from 19.38% to 24.4% (Dhankher et al., 1990). Nugdallah and El
Tinay (1997) reported that G3 – glutelins ranged from 9.0% to 14.0% for cowpea.
2.2.2.4. Insoluble protein:
Insoluble protein (residue) was not extracted because it was linked to the
cell wall (Wall and Paulis, 1978). Residue may be unextracted glutelin plus
variable amounts of globulins and albumins associated with starch and cell debris
(Wilson, 1971). Moreover, a small amount of nitrogen remains insoluble after all
these extraction procedures. This residue consists mainly of proteins form
previously defined groups becoming insoluble due to interaction with lipids,
carbohydrates, or polyphenols via oxidation process (Landry and Moureaux,
1981). Nugdalla and El Tinay (1997) reported that insoluble protein ranged from
0.8 – 3.0% for cowpea.
2.3 Anti-nutrional factors:
48
Anti-nutrients are common in many legumes, including cowpeas. Liener
(1980), has defined these toxic components in legumes as “those causing
physiological response in man or animals when consumed”. These Leguminous
anti-nutrients include protase inhibitors, amylase inhibitors, hemagglutinins,
allergens, aflatoxins, cyanogenic glycoside, favism factors, lathyrogens, metal
binding factors (phytates, oxalates, saponins), anti-vitamins, estrogens,
pressoramins, flatulence factors and polyphenols. Cowpea contain anti-ntrients
such as polypenols, trypsin inhibitors, lectins and phytate (Bressani and Elias,
1978).
49
2.3.1. Chemical nature:
2.3.1.1. Chemical nature of tannins:Tannins are polymeric phenols of higher molecular weight (M.W. 500–
5000 ) containing sufficient phenolic hydroxyl groups to premit the formation of
stable cross–linke with proteins (Swain, 1965).
The main distinctions between the two groups arise from their action
towards hydrolytic agents, particularly acids. The hydrolysable tannins which
have a polyester structure are readily hydrolysed by acids or enzymes into sugars
or related polyhydric alcohols and a phenol carboxylic acid and dependent on the
nature of the later, a subdivision into gallotannins and elogitannins is also usually
made.
Thus on hydrolysis, the gallotannins give gallic acid and the ellagitannins
hexahdroxydiphenic acid isolated normally as its stable dilactone ellagic acid or
acids which can be considered to be derived by simple chemical transformation of
ellagitannins such as oxidation, reduction and ring fission. The condensed tannins
in contrast do not readily break down with acid, instead they undergo progressive
polymerization under the action of acids to yield , the amorphous phlabaphens or
tannin reds (Haslam, 1966). The condensed tannins also referred to as
procyanidins (Weinges et al. 1969) . and formally as leucoantho-cyanidin
(Rossenheim, 1920) because many forms cyanidin upon acid hydrolysis, they are
mostly flavans or polymers of flavan 3-ols (Catechin) and/or flavan 3-4 diols
(leucoanthocyanidins). Both catechin and leucoanthocyanidins are readily
converted by dehydrogenating enzymes or even by very dilute mineral acids at
50
room temperature into flavonoid tannins (Weinges et al. 1968). Heating in acid
solution convert leucoantho cyanidins to the corresponding anthocyanidins and
brown phlobatannins (Swain, 1959). Tannins are insoluble in nonpolar solvents
like ether, chloroform and benzene and are sparingly soluble in ethyl acetate.
Because of the presence of a large number of polar groups, they readily
dissolve in water and alcohol to form colloidal solutions. Extraction of tannin by
aqueous mixture of polar organic solvents depends upon random transfer of
individual bonds from the substrate to the competing sites on the solvent
molecules (Mc-Leod, 1974). Generally most of tannins appear to be found in few
families of the dicotyledons such as the leguminosae (White, 1957). But the
condensed tannins (Structure 1 – 8) are more widely distributed in higher plants
(Schanderl, 1970).
HO
H2O – OO C
HO
HO
COO
O
O
C
O
O
C
HO
HO
HO
HO
HO
COO
HO
HO
COO
O
HO
HO
HO
COO
HO
Structure 1 Glalotannin
HO
COOH HO
OH
HO
HO
HO
HO
HO
COOH
HO
HO
51
OH
OH
OH
OH
HCOO
Structure 3 Hexahydroxy-diphenic
Structure 2 Gallic acid
52
O
HO
Structure 5 Flavanzol (Catechin)
O
HO
Structure 6 Flavan 3-4-diol
(leucoantho-cyanidin)
HO
+
O
HO
Structure 7 3-Hydroxy flavylium
(Anthocyanidin)
HO
O
HO
HO
HO
HO
HO
O
HO
HO
HO
HO
HO
Structure 8 Apossible structure of
grain sorghum condensed tannin
(Haslam, 1977)
HO
O
HO
HO
53
HO
2.3.1.2. Chemical nature of phytic acid:
Beans, in common with other seeds, contain metal salts of myoinositolhexaphosphate or phytic acid . Di–and trivalent metal salts of phytic acid
are relatively insoluble in water. Ferric phytate is nearly insoluble in acid solution
as well as in water. This property has been utilized for the isolation of ferric salts
of phytic acid from acid extracts of natural products (Heubner et al. 1914;
Young, 1936; McCANCE el al. 1935). Quantitative determination of phytic acid
may be based on the analysis of phosphorus or iron in the isolated ferric phytate
(Mc CANCE et al. 1935; Crean et al. 1963; Schormuller et al. 1956).
Alternatively or indirectly, on the determination of the residual iron in solution
after precipitation of ferric phytate from a known concenteration of ferric salt in
acid solution (Crean el al., 1963). Determination of phytic acid based on
phosphorus analysis of ferric phytate has been reviewed by Schormuller et al.
(1956). Wide variations in values for phytic acid were obtained by several
investigators who have used different analytical methods (Schormuller et al.,
1956; Marrese et al., 1961). Phytic acid, the hexaphosphate ester of myo –
inositol (I) , is a major phosphorus storage constituent of most cereals, legumes
and oilseeds (Reddy et al. 1982). Phytic acid has twelve ionisabe hydrogen atoms,
for which dissociation constants have been measured at 28oC by 31P n.m.r
spectroscopy (Costello et al, 1976). The variation of chemical shifts with pH
shows that six of the acidic groups (one from each phosphate) have pka values in
the range 1.1 – 2.1, three others are weakly acidic with pka values between 5.7
and 7.6 and the remaining three lie in the very weak acid range 10.0 – 12.0.
Under physiological pH conditions, therefore, phytic acid is extensively ionised
54
and is capable of interacting strongly with proteins and with metal ions (Reddy et
al. 1982). The phytate – protein interactions are thought to be ionic at low pH and
mediated by cations, through the formation of phytate – cation- protein
complexes, at high pH (Reddy et al. 1982). These interactions lead to reduced
protein solubility and a consequent alteration of their solubility–dependent
properties such a hydrodynamic behaviour, foaming and emulsifying capabilites,
dispersibility in water, etc. Phytate is also a potentially strong ligand and can form
stable complexes. Hence the phosphate groups may chelate as in structure (II) to
give 4–membered ring complexes (Anderson et al., 1977), alternatively two or
more phosphate groups from the same or from different phytate ions may complex
to one metal cation giving structure (III) and the polymeric structure (IV),
respectively, or a phosphate group may serve to bridge two metal ions as in (V)
(Jones et al., 1977). To date there exists little structural information on these
complexes and there is clearly a need for X – ray crystallographic investigations
to fill this void (Kevin et al. 1987).
OPO3H2
OPO3H2
OPO3H2
H2O3PO OPO3H2
OPO3H2
Structure I
O
M
O
O
||
||
(PO3H)5 – inositol – O – P
O –OM – O
P
HO
O
O
P
OH
Insitol – (PO3H)4
Structure (III)
O
55
Structure (II)
2.3.1.3 Chemical nature of the inhibitors:Most of the elicited protease inhibitors are proteins. Trypsin
inhibitors appear to be ubiquitous in all tissues, and they have been
most intensively studied in the legumes and cereals. Soybean seeds
contain two types of trypsin inhibitors, the kunitz inhibitor of 21,000
MW that is specific for trypsin only (1:1 complex) and the Bowman –
Brik inhibitor of 8300 MW that binds independently and
simultaneously to trypsin and chymotrypsin (1:1:1 complex) .Most
legumes contain the Bowman – Brik type inhibitors, with considerable
homology among them, while most legumes do not produce the
Kunitz-type inhibitor. The Kunitz inhibitor, with two disulfide bonds.
About 1h of cooking is required to completely inactivate the Bowman
– Brik inhibitor, unless a reducing compound, such as cysteine, is
added. Several isoinhibitors of typsin are often found in higher plants.
The legume inhibitors are known to be significant, nutritionally,
at least in some animals, (Owen et al., 1996).
2.3.2 Anti-nutritional effect:
2.3.2.1 Anti-nutritional effect of the Inhibitors:
Enzymes are responsible for the myriad reactions associated
with reproduction, growth, and maturation of all organisms. In most
cases, these are desired activities. In some cases, too much enzyme
56
activity, such as polyphenol oxidase– caused browning, can lead to
major losses in fresh fruits and vegetables. In many humans, absence
of or too little of an enzyme is responsible for many genetically
related diseases. Microbially caused diseases present another problem
. The best way to treat these types of diseases is through inhibition of
one or more key enzymes of microorganisms, resulting in their death.
The inhibitors might complete reversibly with substrates or
inhibitor might form acovalent bond with active site groups (affinity
labling inhibitor), or the compound might be treated as substrate and
be catalyzed to product that, while still in the active site.
The last type is the most specific and desirable in medicine and
food because in hibitor can be targeted specially for enzyme.
Enzymes continue to catalyze reactions in raw food materials
after they reach maturity. These reactions can lead to loss of color,
texture, flavor and aroma, and nutritional quality. Therefore, there is
need for control of these enzymes to stabilize the product as food.
Enzyme inhibitors are aslo important in the control of insects and
microorgansims that attack raw food. They also are used as herbicides
in the control of unwanted weeds, grasses and shrubs.
Enzyme inhibitors are an important means of controlling
enzyme activity . An enzyme inhibitor is any compound that decrease
57
intial velocity (Vo) when added to the enzyme – substrate reaction.
There are many enzyme inhibitors, both naturally occurring and
synthetic. Some inhibitors bind reversibly to enzymes and others form
irreversibly, covalent bonds with the enzymes. Some inhibitors are
large proteins or carbohydrates, and others are as small as HCN.
Products of enzyme – catalyzed reactions can be inhibitory.
Change in pH can alter activity by making conditions less
optimum for enzyme activity. Elevated temperatures can decrease
enzyme activity by denaturing some of the enzyme, but at the same
time increasing the velocity of conversion of substrate to product by
the active enzyme. Most enzymologists do not consider either of these
variable to be enzyme inhibitors.
Denaturation of the enzyme eliminates its activity, and this can
be accomplished by shear forces, very high pressures, irradiation, or
miscible organic solvents.
Enzyme activity can also be decreased by chemical
modification of essential active site groups of the enzyme.
Enzymes are aslo inactive when their substrate(s) are removed.
All of these inhibitroy approaches are valid ways of controlling
enzyme activities in foods (Owen et al. 1996). Protease inhibitors in
food are subtances that have the ability to inhibit proteolytic activity
58
of certain enzymes (Liener et al., 1980). They are present in cowpeas
and other legume seeds (Kocchar et al., 1988; Dellagata et al., 1989).
Their importance lies in their possible adverse effect on nutritive value
of plant proteins. Plant breeders in their effort to produce insect
resistant varieties of cowpea have sometimes increased levels of
trypsin inhibitors (Gatehous et al., 1979). These toxicants lower
protein quality by decreasing PER. Protease inhibitors, can be reduced
by soaking and dehulling the seeds followed by heating (Ogun et al.,
1989)
2.3.2.2 Antinutritional effect of the tannin:
These toxicants lower digestibility (Liener 1976), protein
efficiency ratio (PER) and overall nutnitive value of uncooked or
improperly cooked seeds and can cause diarrhea and vomitting (Anon
1976). Cowpeas contain Anti-nutrients such as polyphenols which
may decrease protein digestibility and reduce protein quality
(Bressani et al. 1978). Elias et al. (1979), obtained lower PER for
beans combined with cooking water than for drained beans alone.
They suggested that tannins and/or other pigments interfered with
protein utilization. Elias et al. (1979) also found that tannin
concentration was high in colored seed coats but low in white–coated
seeds. However, Radke et al. (1981) observed that PER values were
59
identical for the white variety of blackeye beans with and without
cook water. Tannin also lowers protein digestibility in cowpeas
(Laurena et al. 1984; 1986). The unfavorable influences of tanins on
nutritional properties of cowpea have bean discussed by Price et al.
(1980). The adverse effects of tannins may be related to the fact that
tannins interfere with protein digestion, affecting digestive action of
trypsin and alpaha amylase either by binding the enzmes themselves
or by binding dietary protein into an indigestible form (Bressani et al.
1982) .Tannins can also be complexed with vitamin B12 causing a
decrease in absorption of the vitamin in rats. Cooking drecreased
tannin and increased in vitro protein digestibility in cowpeas (Laurena
et al. 1984; Uzogara et al. 1990a). Various workers (Akinyele 1989;
Ogum et al. 1989), observed increased losses in tannin when cowpeas
soaked and cooked. Tannin loss may be due to heat degradation of the
tannin molecules or formation of water soluble complexes between
tannin and other tissue molecules of the bean. Such water–soluble
complexes could leach out into the cook liquor. Uzogara et al. (1990a)
observed increased removal of tannins in beans cooked in alkaline
solutions especially under pressure cooking. Tannins form complexes
with proteins, carbohydrates and other polymers in food as well as
with certain metals such as iron under suitable conditions of
60
oncentration and pH (Goldstein et al. 1965). Lease et al. (1969),
reported a marked decreased in blood haemoglobin in rats fed 5%
tannins. They proposed that this phenomenon was due to the
formation of tannin–iron complex which reduce the availability of
iron.
2.3.2.3 Antinutritional effect of phytic acid:
Phytic acid is common in cowpea and other legumes and is the
principal storage form of phosphorus in many dry beans. Phytic acid
occurs as acomplex (phytin with divalent cations or monovalent
cations in discrete regions of the beans and accounts for up to 80 % of
the total phosphorus content (Reddy et al. 1982). Most of the phytates
in dry beans are located in the cotyledons and not in the seed coat.
Anti-nutritonal concern about the presence of phytates in dry bean
arises from the fact that phytate decreases the bioavailabilty of
essential minerals (Ca, Mg, Mn Zn, Fe, Cu) and may posssibly
interfere in the utilization of proteins due to phytate–protein and
phytate–mineral– protein complexes (Oberleas et al., 1981). Under
physiological conditions these complexes may be insoluble thereby
making proteins unavailable for proteolysis in humans and animals. It
has been shown that phytate can inhibit enzymes such as alpha
amylase, pepsin and trypsin under in vitro, conditions (Reddy et al,
61
1982), which may further, reduce substrate utilization. In foods high in
phytate, zinc may not be readily available for absorption since a
phytate: zinc molar ratio of above 20 is reported to be associated with
chemical zinc deficiency (Oberleas et al., 1981).
The hard-to–cook (HTC) defect is a condition whereby bean
cotyledons absorb water but fail to soften during boiling (Stanley et al
1985; Ramcharran et al., 1985; Paredes–Lopez et al., 1989; Hentges
et al., 1991). Development of HTC defect is not well understood and
various factors may cause the defect and leads to increased cooking
time in beans. HTC defect lead to increased consumption of cooking
fuel that may be scarce and expensive in developing countries. This
may limit utilizing cowpeas and other dry grain legumes for
prevention of protein energy malnutrition in these countries. It also
places people who rely on dry beans for dietary protein at a nutritional
disadvantage. The most frequently advanced hypotheses for
explanation of the HTC defect are (a) the middle lamella–pectin–
cation–phytate mechanism of Mattson (1949); (b) the dual enzyme
(phytase + pectin methyl estrase) mechanism of Jones et al., (1983);
(c) the cross- linking of phenolics and / or protein in middle lamella
theory as proposed by Hincks et al. (1987) and (d) the cross – linking
of phenolics and /or protein in middle lamella theory as proposed by
62
Hincks et al. (1987). and Vindiola et al. (1986).; (e) the decreased
solubility of starch and protein theory (Akinyele et al., 1986; Hentges
et al., 1991).
Basically, phytic acid located in the protein bodies of bean
cotyledons chelates divalent cations (Ca, Mg). At high temperature
and high relative humidity conditions in legumes with high moisture
content, there is increased metabolic activity, phytase activation and
membrane degardation. Phytase hydrolyzes phytin in cotyledon cells
to release bound Ca and Mg, which migrate from the cotyledon cells
to the middle lamella. At the same time, pectin methyl esterase (PME)
in the middle lamella hydrolyzes pectin to pectic acid and pectinic
acid and methanol. The divalent cations that migrated to the middle
lamella now react with the released pectinic acid, forming insoluble
Ca or Mg- pectinates that firms the middle lamella and cements cells
together . Decreased pectin solubility and low phytate content have
been correlated to poor cookability in HTC bean (Vindiola et al.,
1986).
63
2.3.3 Anti-nutrients Content:
2.3.3.1 Tannin content of cowpea:
Most plant tissues contain a wide range of secondary products
such as polypenols. The significance of these secondary plant products
is a matter of wids especulation and it has been suggested that some
may serve to protect the plant from pests, diseases and natural
predators (Hulse et al., 1980). Tannic acid contents. Expressed as
percentages of bean dry weight, in raw and processed cowpea
varieties. The ranges are: for raw whole beans, 0.42–0.78%; for
autoclaved beans. 0.33–0.67 % ; for cooked beans, 0.23–0.42% for
soaked beans, 0.37 – 0.69% and for germinated beans,0.29–0.56%.
Cooking and germination decreased tannins contents by 31.0– 47.3
and 23.8 37.0%, respectively. Autoclaving was not as effective as
cooking and germination and losses obtained ranged between 13.8 in
‘kano 1696’ and 38.3% in Nigeria B7 (Anthony et al., 1984).
Two varieties had tannins 2.7 to 25 mg/g raw, of which 7.5 to
48% was destroyed by boiling, 7.4 to 34% (8 varieties) by pressurecooking and 57, 62 or 64% (3 varieties) by soaking for 48 h befor
boiling, which was done for red kidney bean, black cowpea and rice
bean. The varieties with the most tannin white cowpea seemed to be
64
the best of those legumes for feed efficiency, destruction of TIA on
cooking and low tannin content.
Tannin content of cowpea cultivars was 1.24–1.42 mg/g. There
was no significant correlation between IVPD and tannin content (Ene–
obong, 1995).
Tannin content in Red cowpea (RCP) flours ranged from 3.0 to
4.5 mg/g sample. These quality parameters indicated for (RCP), dry
processing generally produced product superior to those obtained from
wet processing (Ningsanond et al., 1989).
It has been hypothesized that part of hard – to – cook (HTC)
defect in cowpeas is due to decreases in solubility and thermal
stability of intraccllular proteins during storage since coagulated
proteins would limit water to starch and pervent full swelling during
cooking (Keshum et al., 1993). Porridges prepared from extruded
millet and press- dried cowpea had high nutritional quality with
acceptable properties for weaning foods (e.g an intermediate
consistency, smooth texure and pleasant color and flavor). Treatment
with sorghum malt allowed the prparation of more fluid products
(Almeida, Dominguez et al. 1993).
Somples of 15 Nigerian cowpea cultivars were analysed. Hotsoaking and cooking significantly reduced trypsin inhibitor. Phytic
65
acid levels were reduced by cold–soaking and hot soaking (Ogun,
1989).
No clear variations in protein quality were observed between
boild and pressure cooked samples. For the biological parameters
Protein Efficiency Ratio (PER), Net Protein utilization NPU & Invitotro protein Digestibility (IVPD) studied raw and cooked chickpea
samples gave the highest values followed by cowpea and peas
(Hashimy et al., 1985).
Studies were conducted on chickpeas (Cicer arietinum), horse
gram (Dolichos biflorus) and cowpea (Vigna sinensis), processed by 6
methods; boiling, pressure cooking, puffing, frying, germination and
germination+cooking. For cowpea most of these treatments improved
IVPD (El Faki et al., 1984).
Study was carried out on 15 local and improved cowpea
genotypes grown in 12 environments, comprising 3 Nigerian locations
over 3 seasons per location. Genotype effects were strongest in
controlling trypsin inhibitor activity while environment was the major
source of variation for tannins, haemagglutinin and phytic acid
contents. Results, indicated that a cowpea genotype grown and
consumed safely in one environment can be poisonous when grown
and consumed in another. Genotype x environment effects were
66
significant for tannins, haemagglutinins and trypsin inhibitor
concentrations (Oluwatosin, 1999).
The cream and speckled African yam bean contains more TI
(30.9 and 25.3 mg/g) than pigeon pea (7.5–14.1 mg/g ) and cowpea
(9.8–20.5 mg/g). Apart from the white cowpea cultivar, they contained
more tannin (1.2–1.4 mg/g) than pigeon pea (0.14-0.97 mg/g) and
African yam bean (0.7–1.2 mg/g). Phytate was lowest in the African
yam bean (6.3–7.5 mg/g) than pigeon pea (8.3–11.3 mg/g) and
cowwpea (8.4–9.9 mg/g). Phytic acid contributed 67.0-74.0% of the
phosphorous in the African yam bean, 66 – 75% in pigeon pea and
54.0–59.0% in cowpea. The IVPD of African yam bean (73.3 ±0.7%)
was lower (P ≤ 0.05) than those of pigeon pea (76.34 ± 0.2%) and
cowpea (77.8 ± 0.4%)
There was a negative correlation between trypsin inhibitor and
IVPD (r = - o.63, p ≤ 0.05). There was no significant correlation
between IVPD and phytate and tannin contents. There was a positive
correlation between protein content and IVPD(r+ 0.69) for the
legumes under study (Ene-Obong, 1995).
2.3.3.2 Trypsin inhibitor activity
TI activities in the raw, autoclaved, cooked, soaked and
germinated cowpea varieties ranged between 19.6 and 28.2 TUI/mg
67
protein (TUI, trypsin units inhibited). Autoclaving and cooking
resulted in a complete loss of activity, while soaking decreases
inhibitor activity by 22.8–42.4%. In the germinated samples,
percentage losses were highest in Igbira and west breed where TI
activites were reduced to 8.4 and 11.0 TUI/mg protein, corresponding
to 59.1 and 57.2% losses, respectively. (Anthony et al., 1984).
The combined effects of soaking, germination and temperature
on cowpea on trypsin inhibitor activity (TIA) was 4.28 mg/g. (Wang
et al., 1997). The inhibitation of human and bovine pancreatic trypsin,
Chymotrypsin and total proteolytic activity by extracts from 5 samples
of red cowpea (Vigna unguiculata) were studied. The thermal liability
of the inhibitors was also assessed . The raw cowpea samples had a
trypsin inhibitor activity (TIA) level of 14.3 mg trypsin inhibited/g
samlpe. Inhibitation of proteolytic activity was influenced by the type
and source of pancreatic enzymes. At all levels of raw cowpea extract
concentration, bovine trypsin was inhibited to a signifcantly greater
extent than was human trypsin. With regards to processing effects,
almost complete inactivation of inhibitors was achieved by cooking
whole cowpea seeds after soaking and dehulling, while only partial
inactivation occurred when raw cowpea was milled into flour before
cooking. It is concluded that effective control of the inhibitory
68
activities in cowpea for maximum nutritional benefits can therefore be
achieved by soaking, dehulling and cooking whole cowpea seeds (Nti
et al., 1996). Trypsin inhibitor (TI) of cultivars of cowpea (Vigna
unguiculata) were 9.8–20.5 mg/g . There was a negative correlation
between TI and IVPD. These legumes may pose no serious problems
to populations consuming them especially when heat treatment is
applied before consumption (Ene-Obong, 1996).
The availability of protein in Italian cultivars of beans
chickpeas, peas lentils, cowpeas an soyabeans was estimated by
quantifying anti-nutritional factors: trypsin inhibitors, tannins, phytic
acid and dietary fibre. Beans and soyabeans had the largest amounts of
trypsin inhibitors, 64.5 and 13.5 UTI/mg, and contained over 1%
phytic acid; lentils had the lowest, 2.4 UTI/mg, and 0.44% phytic acid.
Beans had high tannin contents and had the lowest protein digestibility
value ranging from 71.3 to 78.5%, whereas the other legumes had an
average protein digestibility of 83%. Cooking increased the
digestibility of beans. Protein digestibility was found to be correlated
with tannin and phytic acid contents (Carnovale et al., 1992).
The mature seeds of legumes bought in the philippines were of
11 varieties (species of Psophocarpus, Phaseolus and probably cajanus
and vigna). Protein in dry matter ranged from 20 to 30 % raw and 18
69
to 40% after being boiled until soft, which took 30 min to 2h
according to variety. TIA units were 2 to 41/mg raw, of which 68 to
97% was destroyed by boiling by 76 to 97% for pressure – cooking at
10 lb/in2 for soft, 15 min to 2h. (Beltran et al., 1983).
2.3.3.3 Content of phytic acid:
Germination of cowpea seeds greatly lowered the phytic acid
content. The decrease amounted to 51.6% in Prima and 43.4% in
Adzuki .Soaking decreased phytic acid content of seeds by 19.4 to
280%, and cooking by 7.7–11.7%. Autoclaving had very slight and
maximum loss did not exceed 7.2% in Nigeria B7 (Anthony et al.,
1984; Beltran et al., 1983).
Juice was obtained from 2 cultivars of cowpeas by hot water
extraction, cold water exraction or boiling for 10 min befor cold water
extraction. Trypsin inhibiting units/ml ranged from 4.5 to 6%. Phytic
acid content ranged from 61.8 – 80.5 mg/100g and IVPD ranged from
60.5 – 81.8% for 2 cowpea varieties extraction obtained by hot water
and cold water eraction (Akinyele, 1991). Raw cowpea seeds and
cooked cowpea seeds were analysed for protein, Sulphur amino acids
and trypsin inhibitor (TI). Trypsin inhibitor ranged from 26.7 to 66.2
TI units/mg proteins in raw seeds and from 3.8 to 8.1 TI units /mg
protein in cooked seeds, with the % loss on cooking ranging from 77.7
70
to 92.5%. Protein contents ranged from 20.8 to 26.4% (by wt.). The
study of the 2 varieties indicated that the cotyledon contains most of
the TI and that TI levels in the cotyledon are generally unaffected by
soaking. No correlation was found between TI levels and bruchid (a
storage pest) resistance (Catta et al., 1989).
Phytate content of cowpea cultivars were 8.4–9.92 mg/g. Phytic
acid contributed 54–59% of the phosphorous in cowpea. There was no
significat correlation between IVPD and phytate content (Ene-Obong,
1995).
2.4. Processing:
The appearent insignificant changes in the proximate
composition (DM) with the exception of ether exctract and moisture
content show that the processed were efficient in terms of nutrient
retention (Akiny et al., 1989)
Batches of cowpeas were (i) cooked at 100 degree ºC for approx
1.5h, (ii) germinated in moist cotton wool at room temperature for 72
h; or (iii) fermented for 72 h. Cooking caused slight decrease in phyic
acid and crude protein germination caused considerable decrease in
phytic acid (Akpapuman et al., 1985).
The preparation of cowpeas, by soaking for 1h,
dehulling, cooking for 45 min and sun drying to 15%
71
moisture, destroyed trypsin inhibitor activity almost entirely
(Abbey et al., 1988)
Cowpeas seeds were germinated at 25- 30oC for 24h,
in-vitro protein digestibility was determined, it did not
improve significantly by germination or decortication but was
improved by cooking (Nnanna and Phillips, 1989).
Two cowpea (Vigna unguiculata) variety 3 legume spp.
(Centrosema pubescens, Psophocarpus tetragonolobus, Cajanus
cajan), 3 sorghum (Sorghum bicolor) and 2 millet (Digitara spp).
were autoclaved for 20 min (30 min for winged bean) at 21 1b/inc2.
Cooked products were analysed for tannin contents. Assayable
polyphenol levels were generally higher in cereals than legumes
(1.76–8.21 vs. 0.3–4.4 % as tannins). The dark colored sorghum seed
coats contained the highest levels. In comparison to raw foods cooked
products contained substantially less tannin (up to 71% loss on
cooking). due to the heat destruction and complexation of
protein/tannin. For nutritional purposes light colored sorghum seeds
and moist cooking are recommended (Ekpenyong, 1985). Cowpea is
grown in newly reclaimed lands which are slightly to moderately
saline. Both dry mature seeds and snap pods are popular as food .
72
Effect of growth regulators and reflecting antitranspirant on chemical
composition and nutritional quality of raw and cooked cowpea seeds
after soaking in different media was studied. Foliar application with
gibberellic acid (GA3) and white wash (reflecting antitranspirant,
CaCO3, 6% suspension) especially under saline conditions, improved
the nutritive value of raw cowpea seeds by increasing total protein,
carbohydrate, total free amino acids and in vitro protein digestibility
as well as reducing anti-nutritional factors, i.e trypsin inhinitor
activity, phytic acid and tannins. Cooking in water containing NaCl
(2%) after soaking in hot water for 12 h showed the lowest content of
protein, carbohydrate and antinutritional factors and the highest
content of fibre (Bakr et al., (1991).
The albumin + globulin fraction increased (P ≤ 0.05) for corn
germinated seeds, accompanied by a decrease in the prolamin zein
fraction while the G1-glutelim fraction decreased (P ≤ 0.05) as
reported by (El Khalifia et al., 1996).
The protein content decreased during germination; the albumin
and glutelin, increased by 61.5 and 57.0% respectively, while a 54.6%
decrease was noted in the prolamin fraction. The globulin fraction
increased at the beginning of germination but decreased at the end.
Germination significantly (P ≤ 0.05) increased the crude protein,
73
nitrogen solubility and in-virtro protein digestibility but decreased the
fat, phytic acid and polyphenol contents of the pumpkin (Telfairia
occidentalis) seeds (Giami et al., 1999; El Khalifa and El Tnay, 1999)
reported that albumin, globulin and residue increased significantly (P
≤ 0.05) for two germinated sorghum cultivars for 72h, but the glutelin
decreased significantly (P ≤ 0.05). For chickpea (Cicer arietinum)
germinated seed for 6 days little variation was observed on total
nitrogen content, however, the non protein nitrogen was doubled.
A decrease of 19-1% and 20.6% was obsorved in total globulin
and albumin fractions, respectivaly. The trypsin inhibitor activity
showed a little drop after 6 days of germination, indicating a possible
increase on digestibility of the proteins (Neves et al., 2001).
Suda et al (2000) reported that protein fractions in wild
poinsettia (Euphorbia heterophylla), exhibit different degradation
patterns during germination and initial seedling development; gobulins
being continuously degraded after the start of imbibition whereas saltsoluble fractions were degraded between 36 and 72 hours, and
albumins between 60 and 84 hours. Globulin depletion is
accompanied by increase in free amino acids in the endosperm
whereas intense albumin depletion did not occur. This result suggests
that during albumin depletion there was a rapid transfer of amino acids
74
to the growing embryo. Fiel et al. (2003) reported that the globulin
fractions of cooked faba beans ranged from 36.6 to 55.0% compared
to 69.5-78.1% for the uncooked seeds. The Albumin fractions of
cooked faba beans ranged from 0.6 to 1.0% compared to 1.4- 3.4% for
the uncooked. The prolamin fractions of cooked faba beans ranged
from 2.6 to 5.9 % compared to 2.1–4.1% for the uncooked seeds.
The G1- glutelin fractions of cooked faba beans ranged from 1.5
to 2.3% compared to 0.9–2.2 for uncooked seeds. The G2-glutelin
fractions of cooked faba beans ranged from 4.1–7.8% compared to 1.9
– 6.2 % for uncooked seeds.
The G3–glutelin fractions of cooked faba beans ranged from
27.1 to 45.3% compared to 8.9-14.4% for the uncooked seeds.
Residues of cooked seeds ranged from 2.4 to 3.6% compared to 1.83.4 for the uncooked seeds. Use of germenated cowpeas in food is fast
gaining popularity in Nigeria an other west African countries
(Papunam et al. 1985; Ologhobo et al. 1986; Obizoba 1989).
Germination or sprouting improves proteins, carbohydrates, vitamins,
minerals and overall nutritional values of legumes and leads to
reduction in some toxicants (Vanderstop 1981; Boralkar et al . 1985).
Germination leads to an increase in enzymes (Nnanna and Phillips,
1989) as well as free amino acids. Vitamins such as ascorbic acid
75
riboflavin, niacin, choline and biotin are increased by germination
(Nnanna and Phillips, 1988). However, thiamin and pantothenic acid
did not change while folic acid diminished by germination.
Germination leads to a slight decrease in trypsin inhibitor activity, and
ample decrease in starch and flatulence-causing oligosaccharides in
cowpeas (Ologhobo et al., 1986; Nnanna and Phillips, 1988). During
germination, reducing sugar content increases while polyphenol and
phytate levels are reduced (Chen et al. 1977). There is general
increase in nutritive value of germinated cooked cowpeas (Obizoba
1989). Germination imparts a characteristic agreeable flavor to
cowpeas probably because of amides released during
germination(Kurien 1987). The germinated cowpeas can be dried and
cooked later into sweet savory dishes with good nutritional qualities
(Uzogara et al. 1992).
Cowpea cultivars with the greatest peak viscosities showed low
stabilities to extended cooking. Roasting depressed paste viscosity
properties of all the cowpea cultivars studied. Tannin concentrations
were 0.3 – 6.9 and 7.2 – 116 mg CE/g flour from whole cowpea seeds
and seed coats respectively, increasing with intensity of seed color.
Although dehulling removed 98% of the tannin content of raw
cowpeas, improvement in protein quality as a result of dehulling was
76
observed for only the highly–pigmented in Maroon–red variety.
Roasting significantly improved digestibility. Roasted cowpeas
possess adequate nutritional and functional qualities as protein
supplements in cereal – based weaning foods (Plahar, 1997).
Six commonly consumed legumes in India were subjected to
the following cooking processes: roasting (160oC), germination (24) h,
pressure cooking (15-lb for 15 min), germination and roasting.
Reduction of phytic acid was apparent when roasting (43%) or
germination (36%) were used; pressure cooking did not achieve any
significant differences. Each legume produced different results.
Lentils had the least phytic acid degradation along with the cowpea,
comparatively the moth bean and the bengal gram exhibited the most.
Changes in the solubility of the minerals were larger in the seeds that
had been germinated and then rosted but were not significant (P ≤
0.01). It is concluded that roasting and germination are effective in
reducing levels of phytic acid and enhancing mineral solubility
(Vaishale et al., 1998). Roasting and pressure cooking increased the
net protein ratio values of the cawpea, canavalia and lupin flours
although the values were lower than that for casein. Both treatments
decreased trypsin inhibitor activities. Tannins decreased slightly in
cowpea and by 38 and 68% in canavalia and lupin flours, respectively.
77
Apparent digestibility of lupin protein was similar to that for casein at
roasting temperatures up to 200oC and after autoclaving. Values were
slightly lower for the other flours. True digestibility values were
similar. Available lysine was not significantly affected by treatment.
Jibaja et al. (1988). Wet–heat treatments such simple boiling
(atmospheric preasure, 100oC) and pressurized boiling (higher than
100oC) reduced the polyphenol content of mature dark red seeds of
cowpea (Vigna unguiculata) c.v UPL CP3 by 61 to 80% and increased
protein digestibility in vitro (IVPD) by 6 to 25%. Pressurized
steaming (higher than 100oC) removed 48 to 83% of the polyphenols
but increased IVPD by only 1.1 to 4.2%. Dry heat as exemplified by
roasting and microwave treatment inactivated 58 to 71% of the tannins
but increased IVPD by only 1%. All the heat treatments were effective
in removing or inactivating polyphenol although different IVPD
values resulted (Laurena et al., 1987). Six types of fingermillet
(Eleusine coracana)–based tempe were developed by incorporating
either common beans, groundnuts, cowpeas, mung beans, chickpeas,
sesame and/or their mixtures and fermented by Rhizopus oligosporus.
The proximate and mineral composition was not changed significantly
by fermentation. Tempe processing reduced the tannin and hydrogen
cyanide contents by 55.2–75-7, and 71.0- 86.2% respectively. The in-
78
vitro protein digestibility was improved (Mugula et al. 1999). Soaking
cowpea seeds (25oC, 24h) reduced trypsin inlibitor activity by 20%,
whereas boiling of soaked seeds decreased TIA by 85%. A slight
increase in TIA occurred in soaked cooked and fermentation for 18h
cowpeas (Prinyawiwat kul et al., 1996).
Though soaking significantly reduced the content of tannins
alone in Vigna aconitifolia, total free phenolics and tannins were
markedly reduced in Vigna sinensis. Greater loss of total free
phenolics as well as tannins occurred under autoclaving compared to
soaking and cooking in both legumes investigated. In Vigna
aconitifolia, soaking in distilled water for 6h and cooking for 30 min
reduced the phytic acid content by up to 43%. Maximum reduction in
the level of phytic acid (36%) was observed in distilled water soaking
compared to cooking and autoclaving in Vigna sinensis.
Limited loss in content of phytic acid was noticed under
autoclaving compared to soaking and cooking in four pulses studied.
However, IVPD of Vigna aconifolia and Vigna sinensis was enhanced
to 12.5 and 14.8%, respectively, under autoclaving. Of all the
processing methods, autoclaving seemed to be the most efficient for
reduction in content of the anti-nutrients, except phytic acid.
(Vijayakumari et al., 1998). A decreasing trend was noted in phytic
79
acid and trypsin inhibitor of cowpea flour when processed into milk,
followed by a further reduction at the end of fermentation (Sanni
et al., 1999). All cowpea varieties contained typsin inhibitors, lectins,
hydrogen cyanide, tannins and phytic acid. Autoclaving destroyed all
the trypsin inhibitors and lectins and significantly reduced HCN but
tannic and phytic acids were heat stable. In raw samples, values
obtained for these compounds, in sequence, were 10.9 to 33.7 trypsin
inhibitor/units mg/protein, 28.6 to 76.6 haemagglutinating units/mg
protein, 1.6 to 3.9mg/100g, 0.2 to 0.4mg/g and 422.3 to
543.4mg/100g. respectively. Autoclaved samples contained HCN 0.73
to 1.32 mg/100g, tannins 0.17 to 0.36 mg/g., and phytic acid 314.97
to 420.54 mg/100g. The amount of total phosphorus that remained
bound to phytic acid and was thus unavailable ranged from 29.60 to
33.15%. Antinutritional factor concentrations differed between
varieties (P ≤ 0.001)(Oke et al., 1996). Autoclaving completely
eleminated trypsin in hibitor, haemagglutinin and hydrogen cyanide
(HCN), whereas it reduced (P ≤ 0.01) the levels of phytic and tannins
in the cowpeas. In raw and autoclaved forms, the insect susceptible
cowpeas were better (P ≤ 0.05) utitized by rats than their resistant
counterparts, a condition which was attributable to lower levels of
anti-nutrients in the susseptible varrieties. In the raw cowpeas, trypsin
80
inhibitor, haemagglutinin and HCN were significant variables
affecting cowpea protein utilization, while in autoclaved samples,
tannic and phytic acids were important though nonsignificant factors.
It is concluded that autoclaving at 105oC under 15 psi for 30 min
improved the protein quality of the insect susceptible more than the
insect resistant cowpea varieties (Umoren et al., 1997). Dry heating
and autoclaving reduced the antinutritional components significantly.
The in vitro protein digestibilities of raw, dry–heated and autoclaved
legume seeds (Acacia nilotica) were 61.2, 77.4 and 80.2%,
respectively (Siddhuraju et al., 1996). Germination decreased the
phytic acid content of chickpea (Cicer arietinum L.) and pigeonpea
(Cajanus cajan L.) seeds by > 60% and that of mung bean
(Vignaradiata), urd bean (Vigna mungo) and soybean (Glycine max)
by about 40 % . Fermentation decreased phytic acid content by 26–
39% in all these legumes with exception of pigeonpea in which it was
decreased by 50%. Autoclaving and roasting were more effective in
decreasing phytic acid in chickpea and pigeonpea than in urd bean,
mug bean and soyabean. Germination greatly increased the in vitro
protein digestibility (IVPD). IVPD was only slightly increased by
roasting and autoclaving of all legumes. Germination and fermentation
also decreased the total dietary fibre (TDF) in all legumes.
81
Autoclaving and roasting caused slight increase in TDF values. All
the processing treatments had little effect on calcium, magnesium and
iron contents (Chitra et al., 1996). Dry heating as well as autoclaving
significantly reduced anti-nutritional factors ofthe little–known
legume, velvet bean (Mucuna pruriens L. DC.) .Protein efficiency
ratio, true protein digestibility, biological value, net protein utilization
and utilizable protein were significantly improved by autoclaving
compared with dry heating, the values of true protein digestibility and
net protein utilization of dry–heated samples were significantly higher
than the raw samples (Perumal– Siddhuraju et al., 1996).
Soaking for 12h was most effective in reducing cooking time,
tannin and phytate concentrations; it also improved IVPD. Prolonged
soaking (24h) decreased calcium and iron values by 19 and 35%,
respectively. Dehulling showed that Ca, Fe, Mg and Zn were
concentrated in seed coat of African yambean. Dehulling reduced
tannin content but had no significant effect on phytate content or in
vitro protein digestibility except for seeds soaked for 12h before
Dehulling for 24h increased crude protein content by 16 % (P ≤ 0.05) .
Blanching and roasting increased IVPD by 8–11% . Fermentation had
no effect on crude protein, Ca Fe. Mg or Zn but significantly reduced
phytate content. Fermentation had no advantage over heat treatment
82
with respect to improving IVPD (Ene- Obong et al., 1996). NaHCO3
solution and autoclaving significantly reduced the contents of total
free phenolics (85-88%) compared to raw seeds. Autoclaving (45 min)
reduced tannins by up to 72%. Soaking seemed to have limited effect
in eliminating phytohaemgglutinating activity whereas autoclaving
(45 min) seemed to eliminate the haemagglutinating activity
completely. The reduction in phytic acid was found to be some what
greater in distilled water soaking (28%) compared to NaHCO3
solution (22%). Only a limited loss in phytic acid was observed under
cooking as well as autoclaving. Loss of HCN was greater under
autoclaving (87%) compared to the other processes studied. Of the 3
sugers analysed, soaking reduced the level of verbascose more than
that of stachyose and raffinose. Autoclaving reduced the content of
oligosacccharides more efficiently (67–86%) than ordinary cooking
(53–76%) . Autoclaving improved the in vitro protein digestibility
(IVPD) significantly (13%). Of all the different water and
hydrothermal treatments studied autoclaving seemed to be the most
eficeient method in improving IVPD and eliminating the anti-nutrients
investigated except phytic acid (Vijayakumari et al., 1995).
Fermentation of various rice and bengal gram dhal blends prepared by
mixing them in different proportion at 35oC for 18h brought about a
83
significant decline in phytic acid content to extent of 23 to 36% over
the control. Incorporation as well as fermentation improved the starch
and protein digestibility (in-vitro) of all the rice – bengal gram dhal
mixures. Improvement in starch and protein digestibility is related to
the reduction in phytic acid content, as this anti-nutrient is known to
inhibit amylolysis and proteolysis, A significant negetive correlation
was found between phytic acid and digestibility of stach and protein
(Anshu – Sharma et al., 1995).
Anti-nutnitional factors such as lectins where higher in raw and
boiled nunas (Phaseolus vulagaris) samples than in roasted nunas,
while tannin levels did not change from raw to roasted treatment.
Overall, in vitro digetibility was slightly lower for toasted nunas than
boiled dry bean (Beem et al., 1992) . The application of dry heat to the
Red gram (pigeonpea) seeds and meal was not effective in inactivating
the trypsin inhibitory activity (TIA) and chymotrypsin inhibitory
activity (CIA). Soaking for 24h followed by cooking for 20 min was
effective in distroying the TIA and CIA (Mulimani et al., 1994).
Indigenous fermentation of coarsely ground dehulled black- gram dhal
slurry at 25,30 and 35oC for 12 and 18 h reduced the concentrations
of phytic acid and polyphenols (P ≤ 0.05) The unfermented legume
batter had high amounts of phytic acid (1000 mg/100g) and poly
84
phenols (998 mg/100g), and these were reduced to almost half in the
protuct fermented at 35oC for 18h. In vitro digestibility of starch and
protein was improved (P ≤ 0.05) with increase in the temperature and
period of fermentation. A significant and negative correlation was
found between the in vitro digestibility and anti-nutrients contents
further strengthens these findings (Yadav et al., 1994). Four Kabuli
type varieties were evaluated, the trypsin inhibitor activity (TIA) of
the varieties was reduced by germination or by cooking the seeds.
Reduction in TIA on sprouting in the variety ILC 116 was
significantly lower than in the other varieties, but this characteristic
was not seen when it was cooked. True digestibility of the protein
correlated negatively with TIA and positively with protein content
(Savage et al., 1993). Processing, especially presoaking followed by
boiling in water or in alkaline solutions at atmospheric pressure,
reduces the phytic acid content of cowpeas (Ogun et al. 1989;
Uzogara et al. 1990a) while pressure cooking or autoclaving caused
less loss of phytic acid (Uzogara et al. 1990a; Ologhobo et al. 1984).
85
CHAPTER THREE
MATERIALS AND METHODS
86
3. MATERIALS AND METHODS
3.1 Material:
3.1.1 Cowpea seeds:
The dry cowpea seeds (Vigna unguiculate L. Walp) of the two cultivar
were obtained from El Obeid Research Station.
3.1.1.1 Chemicals and Reagents:
Potassium sulphate, cupric sulphate, sulphuric acid, sodium hydroxide,
boric acid, hydrochloric acid, methyl red, ethanol (EOH), petroleum ethen (b.p.
60–80ºC) soddium chloride, isopropyl alcolhol (PrOH), sudium borate, sodium
dodecyl sulphate (SDS), 2-mercaptoethanol (2-ME), vanillin, methanol, catechin,
potassium hydroxide sodium carbonate, trichloroacetic anid, casein, sodium
dihydrogen phosphate, sodium hydrogen phosphate, potassium thiocyanate,
ferrichitrate, hexane, formaldehyde, sodium azide, bromocersolgreen. All
chemicals used in the study were of analytical grade.
3.1.2 Apparatus:
Centrifuge, (PTL limted, London), Gerhardt lieraeus shaker,
spectrophotometer, (Unicam Instruments), Cambridge water bath, Heraeus Hanau
mettler sensitive balance, Soxhelt extracting apparatus, magnetic stirrer, steamed
out apparatus, oven,mill, autoclave, heater, incubator, keldahl apparatus,
thermometer, burette, pipette, volumetric flasks, beakers, conical flasks,
measuring cylinders, test tubes, Bunsen burner, aluminum foil, stand , funnel,
filter paper (Whatman No.1), pH meter, deep freezer were utilized.
3.2 Methods:
87
3.2.1 Preparation of cowpea samples:3.2.1.1 Cleaning
About 5 kg of mature and dry seeds of cowpea (Vigna unguiculata L.
Walp) were obtained from El Obeid Research station and were cleaned
thoroughly; freed from foreign matter, broken seeds, and immature seeds. The
seeds were stored in plastic containers at room temperature.
3.2.1.2 Autoclaving:
About 2 kg of seeds were autoclaved at 15 1b, or 20 1b for 15, 30, 45 minOne hundred seeds (100) were placed in 500 ml conical flask, 150 ml. distilled
water were added to the flask. Then flasks were placed in the autoclave. After
autoclaving, liquied was separated, and the autoclaved seeds were dehydrated in a
hot air oven at 50 oC to constant weight and were powdered, defatted and kept for
further use.
88
3.2.1.3 Roasting:
About 2 kg of seeds were roasted at 90oC or 120oC for 30 , 45 and 60
min., the roasted seeds were powdered by micro hammer mill with 0.5 mm mesh
size. The flour was placed in a conical flask and n-hexane was added, the solvent
to flour ratio was 10:1. The mixture was stirred for 16 h and was then filtered and
washed again with n- hexane and was air dried overnight at room temperature and
kept in clean bottles for further use.
3.2.1.4 Germination of Cowpea samples:
Germination was carried out according to the method of Bhise et al.
(1988) .Broken grains were removed by hand. The seeds were soaked with about
3 volumes of distilled water overnight at room temperature, with two changes of
water during the day to remove dirt and extra husks. The wet grains were then
soaked in 1-2 volumes of 0.2% formaldehyde solution for 40 min to retard mold
growth during germination. The soaked grains were then washed with distilled
water several times and soaked in water for 20 min to remove residual formaldehyde. These seeds were spread evenly (about 1 cm thick) in plastic tray,
with plenty of air space, covered with cheese cloth and germinated in an air
circulating incubater at 30 ± 2oC for 1, 2, 3, and 4 days. Water was sprinkled on
the grains every 12h to avoid drying; non germinated and moldy seeds were
discarded. Sprouted cowpea samples were dried at 50 oC to constant weight and
finely ground. Samples were taken to fractionate the protein on the basis of
solubility, and to determine in-vitro protein digestibility (IVPD), phytate and
trypsin inhibitor acctivity (TIA)
89
3.2.1.5 Cooking of sprouted cowpea samples:
Samples were cooked according to the mothod of ICAITI (1978). One
hundred seeds were placed in 150 ml boiling flask and boiled in distilled water for
45 min. Cooked seeds were dried at 50oC to constant weight and finely ground.
Samples were taken to fractionate the protein on the basis of solubility to
determined IVPD, phytate and trypsin inhibitor activity (TIA)
3.3 Analytical methods:
3.3.1 Proximate analysis:
Proximate analysis was determined using the official methods (AOAC,
1990) . The nitrogen contents of the flours were determined using micro Kjeldahl
method (Pearson, 1970).
3.3.2 Protein fractionation:Cowpea proteins can be separated into five fractions by selective
extraction method (Landry and Moureaux, 1970, 1976, 1981, 1982). 3.5g. defatted
samples were kept in suspension with 35 ml extractant by magnetic stirring in
50ml centrifugal tubes. The duration and number of extractions with solvent and
identification of protein fractions were carried out as follows:
Step I: The first fraction (globulin)
The powdered sample (3.5g) was mixed with 35ml of 0.5M NaCl for 3
extraction times (60,30, 30 min) at 4oC. The total supernatants collected were 105
ml
Step II: The second fraction (albumin):
90
The residue was mixed with 35 ml of distilled water for 2 extraction times
(15, 15 min) at 4oC. The total volume collected was 70 ml.
Step III :- The third fraction (prolamins):
The resdiual material was stirred with 60% ethanol twice for 30 min at
20oC and then at 60oC for 30 min, followed by extraction with 55% isopropanol
at 20oC three times (60, 30, and 15 min. with stirring).
The total volume collected was 210 ml.
Step IV: The fourth fraction (G1–glutelins):
The residue was extracted with 60% ethanol plus 0.6 % 2mercaptoethanol (2-ME) and stirred twice for 30 min (20oC), then extracted with
55% PrOH containg 2-ME (0.6%) at 20oC twice for 30 min. The total volume
collected was 140 ml.
Step. V:- The fifth fraction (G2- glutelins):
Borate buffer, pH 10 (0.0125 M N2aB4O7 – 10H2O and 0.02 M NaOH). 2g
of sodium borate were dissolved in 250 ml distilled water and 0.4g of sodium
hydroxide in 250 ml distilled water were added and minxed together, and was
adjusted to pH 10 using either NaOH or HCl) with 0.6% 2.ME and 0.5 M NaCl
were used with stirring for 60,30 and 30 min (20oC ). The total volume collected
was 105 ml.
Step VI:- The sixth fraction (G3- glutelins),
Borate buffer, pH 10 with 0.6% (2-ME) 2- mercaptoethanol and
0.5%sodium dodecyl sulphae (SDS) were used with stirring for 60, 30, and 15
min (20oC). The total volume collected was 105 ml.
91
Fractions I and II contained the albumins and globulins, the free amino
acids and small peptide fragments. Fraction III contained the prolamin. Fraction
IV contained G1- glutelins. Fraction V contained G2 –glutelins. Fractian V
contained G3 – glutelins. The solid material was isolated from extractants by
centrifugation at 30000 g for 15 min. For each solvent, supernatants were
combined to give the total extract. The nitrogen content of each of these six
fractions was determined by mircro–Kjeldahl method. The residue left after
extraction was also analysed for nitrogen content (Table 1).
92
Table 1 . Protein Extraction Procedure for sequence Ao and Bo
Step
Total Volume
(extract)
Extractant
1
105ml
Nacl,O,5Mwater (4oC)
2
3
4
70 ml
105 ml
105 ml
70 ml
70 ml
5
105 ml
6
105ml
7
o
Water (4 C)
EtOH,60%(20oC)
Time of Extraction
No. of
Protein
(min)
Fraction
groups
60,30,30,
I
Globulins
15,15
II
Albumins
III
Prolamins
IV
G1-glutelins
60, 30, 30
V
G2-glutelins
60,30,15
VI
G3-glutelins
------
-----
30.30.30
And then at 60oC
60,30,30
2-Pr.OH 55%(20oC)
EtOH,60%±2ME O.6%(v/v)(20oC)2PrOH,55%±2MEO.6% (v/v)(20oC)
NaCl,0.5M,PH10±2ME.
0.6% (v/v)(20oC)
NaDodSO4,0.5%(W/v)
PH10±2ME 0.6%(v/v)(20oC)
-------
93
30.30
30,30
Insoluble
Proteins
Calcultions:
Soluble protein% =
Protein fraction% =
T.F × N × TV × 14 × 6.25 × 100
a × b × 1000
So lub le protein × 100
total protein of sample
Where:T.F:
Titre
N: Normality of HCl.
TV: Total volume of the aliquot extracted.
14: Each ml of HCl is equivalent to 14 mg nitrogen.
A:Number of ml of aliquot taken for digestion: (10)
B: Number of g sample extracted : (3.5 g).
1000: To convert from g to mg.
2.3.1.3. In-vitro protein digetibility (IVPD):
IVPD was carried out according to Saunder et al , 1970). A 0.2g of the
sample was placed in a 50ml centrifuge tube, 15 ml of 0.1 N HCl containing
1.5mg pepsin, were added and the tube was incubated at 37oC for 3h. The
suspension was then neutralized with 3.3ml , 0.5 N NaOH, then treated with 4 mg
of pancreatin in 7.5 ml of 0.2 M. phosphate buffer (pH= 8.0), containing 0.005 M
sodium azide; the mixitre was then gently shaken and incubated at 37oC for 24h.
After incubation the sample was treated with 10 ml, 10% trichloroacetic acid
(TCA), and centrifuged at 50.000g, for 20 min at room temperature. Nitrogen in
94
the supernatant was estimated using the micro Kjeldahl method. Digestibility was
calculated using the formula:
Protein Digestibility % =
Nitrogen in sup ernatnt
× 100
Nitrogen in sample
The mixed samples were prepared as follows:
Assuming 25.6% protein in (A) samples and 24.6% protein in (B) samples
taking 0.39g and 0.41g sample containing 0.1g protein for digestion. Theefor the
mixed samples of raw material with treated samples would be carried out as the
below sequences:
1. Ain Elgazal (A) Cultivar
Sample weight =
0.1 × 100
= 0.39g
25.6
(Sample weight)
(Protein content)
Auto/Roasted
Raw
Total protein
0.384g
+
0.00g
0.1g
A1
0.336g
+
0.048g
0.1g
A2
0.288g
+
0.096g
0.1g
0.00g
+
0.384g
0.1g
A4
0.048g
+
0.336g
0.1g
A5
0.096g
+
0.288g
0.1g
A6
0.192g
+
0.192g
0.1g
A0
A3
Control
Control
95
2- BUFF (B) Cultivar,
Sample weight =
0.1 × 100
= 0.41g
24.6
(Sample weight)
(Protein content)
Auto/Roasted
Raw
Total protein
0.4g
+
0.00g
0.1g
B1
0.3g
+
0.1g
0.1g
B2
0.35g
+
0.05g
0.1g
0.00g
+
0.4g
0.1g
B4
0.05g
+
0.358g
0.1g
B5
0.1g
+
0.3g
0.1g
B6
0.2g
+
0.2g
0.1g
B0
B3
Control
Control
96
3.3.4 Determination of tannin in raw and treated samples:
Quantitative estimation of tannin for each sample was carried out using the
modified vanillin-HCl in methanol method as described by Price et al. (1978).
The specific reagent for the determination encompasses equal volumes of 1%
vanillin in methanol (w/v) and 8% conc. HCl in methanol (v/v). These are to be
mixed just prior to use and were rejected when ever a trace of colour appears. The
manipulation is in the sequence of: 0.2g of the ground sample placed into a 100
ml conical flask. Ten ml of 1% HCl in methanol (v/v) were added, shaken for 20
min using mechanical shaker and centrifuged at 2500 rpm. One ml of the
supernatant was pippetted into a test tube and 5 ml of vanillin HCl reagent were
added. The optical density was read using a colorimeter (Lab System Analyzergfilters, J. Mitra and Bros. Pvt. Ltd.) at 500 nm after 20 minutes incubation at
30oC. For zero setting of the colorimeter 1 ml of ablank (1% HCl in methanol).
solution was mixed with 5 ml 4% HCl in methanol (v/v) and 5ml vanillin – HCl
reagent in a test tube. The bank mixture was incubated at 30oC for 20 min.
Calculations:
C E% =
C × 10 × 100
=C×5
200
9
Where:
C = Concentration corresponding to optical density.
10 = Volume of extract in ml
200= Sample weight in mg.
The standard curve of tannin:
97
The standard curve of tannim was traced via a procedure in which 100 mg
D(+) – catechin has been added to 50 ml 1% HCl in methanol in 50 ml volumetric
flask, giving a concentration of 2 mg catechin per 1 ml. A series of 0.0, 0.02, 0.04,
0.06, 0.08, and 0.1mg catechin / ml solutions were prepared according to the plan.
Aliquots of 0-ml , 0.5ml, 1.0 ml, 1.5 ml, 2.0 ml,and 2.5 ml were pippetted
from the stock solution into a series of 50ml volumetnic flasks. Then 1% HCl in
methanol were added up to mark. 1 ml portions from each volumetric flask of the
different concentrations were taken into test tubes, and five ml vanillin – HCl
reagent were added. The optical density was read in a colorimeter at 500 nm after
20 minutes incubation at 30oC from addition of the reagents. For zero setting 1ml
blank solution was mixed with 5ml 4% HCl in methanol and 5 ml vanillin-HCl
reagent in test tube, and was incubated at 30oC for 20 minutes. A linear
relationship between catchen concentration and optical density obtained as shown
in Fig. 1.
3
Optical density at 500nm
2.5
2
1.5
1
98
3.3.5 Determination of phytic acid:
The determination of phytic acid was carried out according to the method
of Wheeler and Ferrel (1971) . One gram of finely ground sample was weighed
into a 125 ml conical flask, extracted with 50 ml 3% TCA (w/v) for 30 min with
mechanical shaking. Then the suspension was centriguged at 3000 rpm. Ten ml
aliquots of the supernatant were transferred into 50 ml boiling tubes. Then 4ml of
FeCl3 were added (2mg ferric iron per ml 3% TCA), centrifuged at 3000 rpm for
15 minutes and the clear supernatant was decanted carefully. The precipitate was
then washed twice by despersing well into 25 ml 3% TCA heated in boiling water
bath for 5- 10 minutes and
Then centrifuged. Washing was repeated once with water. The precipitate was
cautiously dispersed in few ml distilled water enriched with 3ml 1.5 N NaOH with
mixing. The volume was made approximately to 30 ml with distilled water and
heated in boiling water bath for 30 minutes. The content of the tube were filtered
hot (quantitatively) through filter paper (Whatman No.1) and the filtrate was
discarded. The precipitate was dissolved with 40 ml hot 3.2 N HNO3 into a 100
ml volumetric flask . The filter paper was washed with several portions of
distilled water, collecting the washings in the same flask. The contents in the flask
were cooled to room temperature and diluted to volume with distilled water. Five
ml aliquots were transferred to another 100 ml volumetric flask and diluted to
approximately 70 ml with distilled water. Then, 20ml of 1.5M (potassium
thiocyanate) KSCN were added, completing the volume up to the mark. The
intensity of color was immediately assessed (within one minute) in a colorimeter
99
(Lab System Analyzer. 9 giters, J Metra and Bros. Pvt Ltd.) at 480 nm .A blank
probe was run with each set of samples.
Calculation:
The iron content was calculated from a prepared Fe (NO3)3 standard curve.
Then the phytate phosphorous (P) was calculated from the iron (Fe)(of a 4:6 Fe: P
molecular ratio). Standard curve of phosphate was plotted from a procedure
including: 0.4321g of ferric nitrate (Fe(NO3)3) dissolved in distilled water in 1
liter volumetric flask up to the mark. Five mllileters of this solution were taken
into 50 ml volumetric flask and the volume made up to the mark with distilled
water, giving concentration of 10 ppm of ferrous (Fe). Concentrations of 0, 1, 2, 3,
4 , and 5 ppm were preparded by taking 0, 5, 10, 15, 20, and 25 ml from 10 ppm
ferous solution into a series of 50 ml volumetric flasks. Then distilled water was
added up to the mark. Five mllileters aliquots from the standards were pippetted
into a 100 ml volumetric flasks, and diluted up to 70 ml with distilled water. Then
20 ml of 1.5 M KSCN were added , completing the volume with distilled water.
The density of color was read at 480 nm immediately (within one min) in a
colorimeter (Lab System Analyzer 9 filters, J. Mitra and Bros. Pvt. Ltd.). A
standard curve was obtained by plotting concentrations against the corresponding
readings of colorimeter giving a straight line. The phytic phosphorus was
calculsted from the ferric ion concentration assuring 4 : 6 iron : phosphorus molar
ratio.
MI standard ml
Fe+3 conc. ppm
A
Conc/A = K
5
1
0.09
11
10
2
0.18
11
100
15
3
0.20
15
20
4
0.28
14.25
25
5
0.30
12.25
K = 13.6
Phytic % =
6 A × mean k × 20 × 10 × 50 × 100
×
4
1000 × 2
Where:
A = optical density
K=
Con
A
30
Absorbance at 480 nm
25
20
15
10
101
5
= 9648.0 × A
3.3.6. Determination of trypsin inhibitory activity:
The extract of the samples was obtained by shaking 4g of the sample with
40ml of 0.1 M phosphate buffer pH 7.5 (16ml of 0.2 M NaH2PO4 and 84 ml
Na2HPO4 (0.2M) diluted to 200ml) for 3 hours using a mechanical shaker
(Shutted machine R010) at room temperature. The extracts were then centrifuged
at ambient temperature at 3000 rpm. for 20 minutes. The supernatants were
diluted two times and then used for analysis.
For the assay of enzymatic activity, the casein substrate was used for
determining trypsin inhibitor activity in the crude preparations (phosphate buffer
extracts).
A 2% casein solution in phosphate buffer (0.1M pH 7.5) was used as
substrate, while the enzyme used as trypsin (Beef pancreas), 5 mg/ml. The
incubation mixture consisted 0.5 ml trypsin. 2ml 2% casein, 0.9 ml phosphate
buffer (0.1M pH 7.5) 0.4 ml HCl solution (0.001 M) and 0.2 phosphate buffer
extract. In all cases the total volume of incubation mixture were kept at 4 ml.
Incubation was carried out at 37ºC for 20 minutes, after which 6.0 ml of TCA
solution was added to stop the reaction and corresponding blanks were run
concurrently.
In this method one trypsin unit (TU) is arbitararity defined as an increase
of 0.01 absorbance unit at 280 nm in 20 minutes for 10 ml of reaction mixture
under there conditions and the trypsin inhibitor activity as the number of trypsin
units inhibited (Ray and Ras, 1971).
3.3.7 Statistical analysis:
102
Each sample was analyzed in triplicate and the figures were then averaged.
Data were assessed by analysis of variance (ANOVA), (Snendecor and Cochran,
1987) and by Duncan’s multiple Range Test (DMRT) with a probability p ≤ 0.05
(Dumcan, 1955).
103
CHAPTER FOUR
RESULRS AND DISCUSION
104
4. RESULRS AND DISCUSION
4.1 Proximate composition of cowpea seeds:
4.1.1. Moisture content:
The chemical composition of cowpea seeds is shown in Table 2.
The results are expressed as on dry matter basis (DMB). Data obtained
showed that, moisture content for Ain Elgazal cultivar raw uncooked
seeds, uncooked 24h germinated seeds, uncooked 48h germinated
seeds, uncooked 72h germinated seeds, uncooked 96h germinated
seeds, raw cooked seeds, roasted seeds, autocalved seeds, cooked 24h
germinated seeds, cooked 48h germinated seeds, cooked 72h
germinated seeds, and cooked 96h germinated seeds were 5.9%, 4.9%,
4.5%, 4.4% 4.4%, 4.8%, ,4.7%, 4.6%, 4.6%, 4.5%, 4.5%, and 4.4%
respectively. Similarly for Buff cultivar raw uncooked seeds,
uncooked 24h germinated seeds, uncooked 48h germinated seeds,
uncooked 72h germinated seeds, uncooked 96h germinated seeds, raw
cooked seeds, roasted seeds, autocalved seeds, cooked 24h germinated
seeds, cooked 48h germinated seeds, cooked 72h germinated seeds,
and cooked 96h germinated seeds were 5.5%, 4.8%, 4.6%, 4.5% 4.5%,
5.2%, 4.8%, 4.6%, 4.5%, 4.6%, 4.5% and 4.4% respectively. Data
obtained showed that germination and heat treatment significantly
(P ≤ 0.05) decreased, moisture content. This disagreed with that
reported by Akinyele (1989).
105
Table 2. Proximate composition of some cowpea preparations
Cultivar
Treatment
Ain Elgazal cultivar
Moisture %
Protein
%
Ash
%
Fiber
%
Oil
%
Moisture %
Protein %
Ash
%
Fiber %
Oil %
Raw uncooked seeds
Germinated 24 h. (uncooked)
Raw cooked
Roasted seed
Autoclaved seeds
Germinated 24 h. (cooked)
5.9±0.1a
25.6±0.01e
3.2
2.5
1.5
5.5±0.01a
24.6±0.01e
3.5
2.2
1.6
4.9±0.2b
26.2±0.02c
3.4
2.6
1.5
4.8±0.02b
25.0±0.0d
1.5
c
4.5±0.01
c
4.4±0.01
c
4.4±0.00
c
28.6±0.1
b
30.7±0.2
a
3.6
3.8
4.0
2.7
2.9
3.0
1.6
1.6
4.6±0.02
d
4.5±0.01
d
4.5±0.01
2.3
1.6
27.4±0.01
3.7
2.4
1.6
28.4±0.01
b
3.8
2.5
1.6
30.6±0.01
a
3.9
2.6
1.7
de
4.0
2.8
1.7
4.0
3.1
1.6
5.2±0.01
23.6±0.02
4.7±0.1b
25.9±0.01c
4.0
3.1
1.6
4.8±0.1a
24.0±0.02d
4.0
3.0
1.7
c
26.0±0.02
c
c
4.0
3.0
1.7
26.2±0.02
c
4.6±0.0
25.9±0.01
4.0
4.6±0.03
c
4.5±0.01
d
27.5±0.02b
4.5±0.01
d
b
4.2
31.0±0.01a
4.3
4.4±0.01e
28.0±0.01
4.1
c
4.2
3.1
3.0
3.2
1.6
1.6
a
3.6
c
c
4.8±0.2
a
27.5±0.0
c
Buff cultivar
c
4.6±0.01
d
4.5±0.00
c
25.0±0.01
25.0±0.0
c
4.2
3.1
1.7
27.4±0.03
b
4.2
3.1
1.7
ab
4.3
3.1
1.7
4.3
3.1
1.7
1.6
4.6±0.03
3.2
1.6
d
4.5±0.1
28.4±0.01
3.2
1.6
4.4±0.01e
28.9±0.00a
Values are means (±SD).
Means not sharing a common superscript letter in a column for the same cultivar are significantly different at (P < 0.05) as
assessed by Duncan's Multiple Range Test.
106
4.1.2 Protein Content:
Data obtained showed that protein content for Ain
Elgazal cultivar raw uncooked seeds, uncooked 24h germinated seeds,
uncooked 96h germinated seeds, raw cooked seeds, roasted seeds,
autocalved seeds, cooked 24h germinated seeds, cooked 48h
germinated seeds, cooked 72h germinated seeds, and cooked 96h
germinated seeds were 25.6%, 26.2%, 27.5%, 28.6%, 30.7%, 25.9%,
25.9%, 26.0%, 26.2%, 27.5%, 28.0% and 31.0% respectively. For
Buff cultivar raw uncooked seeds, uncooked 24h germinated seeds,
uncooked 96h germinated seeds, raw cooked seeds, roasted seeds,
autocalved seeds, cooked 24h germinated seeds, cooked 48h
germinated seeds, cooked 72h germinated seeds, and cooked 96h
germinated seeds were 24.6%, 25.0%, 27.4%, 28.4%, 30.6%, 23.6%,
24.0%, 25.0%, 25.0%, 27.4%, 28.4% and 28.9% respectively. Data
obtained showed that germination and heat treatment significantly
(P ≤ 0.05) increased, protein content. These results reported by
Akpapuman et al., (1985). And Naves et al. (2001), bur were similar
to those reported by Giami et al. (1999); Obong, (1995); Vanderstop
107
(1981; Boralk et al. (1985); Alonso et al. (1999), Akinyele (1989);
Perumal-Siddhuraju et al. (1996).
4.1.3. Ash content:
Data obtained showed that ash content for Ain Elgazal cultivar
raw uncooked seeds, uncooked 24h germinated seeds, uncooked 48h
germinated seeds, uncooked 72h germinated seeds, uncooked 96h
germinated seeds, raw cooked seeds, roasted seeds, autocalved seeds,
cooked 24h germinated seeds, cooked 48h germinated seeds, cooked
72h germinated seeds, and cooked 96h germinated seeds were 3.2%,
3.4%, 3.6%, 3.8%, 4.0%, 4.0%, 4.0%, 4.0%, 4.1%, 4.2%, 4.2% and
4.3% respectively. For Buff cultivar raw uncooked seeds, uncooked
24h germinated seeds, uncooked 48h germinated seeds, uncooked 72h
germinated seeds, uncooked 96h germinated seeds, raw cooked seeds,
roasted seeds, autocalved seeds, cooked 24h germinated seeds, cooked
48h germinated seeds, cooked 72h germinated seeds, and cooked 96h
germinated seeds were 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.0%,
4.0%, 4.2%, 4.2%, 4.3% and 4.3% respectively. Data obtained
showed that processing significantly (P ≤ 0.05) increased, ash
content, a results which are similar that reported by Giami et al.
(1999), Obong (1995) Vanderstop (1981), Boralk et al. (1985);
Alonso et al. (1999), Akinyele (1989).
108
4.1.4. Fibre content:
Data obtained showed that fibre content for Ain Elgazal cultivar
raw uncooked seeds, uncooked 24h germinated seeds, uncooked 48h
germinated seeds, uncooked 72h germinated seeds, uncooked 96h
germinated seeds, raw cooked seeds, roasted seeds, autocalved seeds,
cooked 24h germinated seeds, cooked 48h germinated seeds, cooked
72h germinated seeds, and cooked 96h germinated seeds were 2.5%,
2.6%, 2.7%, 2.9%, 3.0%, 3.1%, 3.1%, 3.1%, 3.0%, 3.2%, 3.2% and
3.2% respectively. For Buff cultivar raw uncooked seeds, uncooked
24h germinated seeds, uncooked 48h germinated seeds, uncooked 72h
germinated seeds, uncooked 96h germinated seeds, raw cooked seeds,
roasted seeds, autocalved seeds, cooked 24h germinated seeds, cooked
48h germinated seeds, cooked 72h germinated seeds, and cooked 96h
germinated seeds were 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.8%, 3.0%,
3.0%, 3.1%, 3.1%, 3.1% and 3.1% respectively. Data obtained
showed that processing significantly (P ≤ 0.05) increased, fibre
content, these results were different from those reported by Akinyele
(1989) but similar to that reported by Perumal-Siddhuraju et al.
(1996).
109
4.2.5 Oil content:
Data obtained showed that oil content for Ain Elgazal cultivar
ranged from 1.5% to 1.6% and for Buff cultivar ranged from 1.6% to
1.7% that showed insignificant (P ≤ 0.05) increased in oil conttent for
both cultivar. These results were different from those reported by
Akinyele (1989).
4.2. Protein fractions:
4.2.1. Globulin fractions:
As shown in Tables 3, 4 and 5, raw germinated, raw germinated
cooked, autoclaved and roasted seeds were fractionated.
The major protein fraction globulin showed significant (P ≤
0.05) increase they were 87.5%, 80.6%, 76.4%, 71.4%, and 70.2% for
Ain Elgazal raw, the first, second, third, and fourth day respectively.
Similarly for Buff they were 89.8%, 81.3%, 78.3%, 75.6% and 72.7%
respectively, a result which are similar to that reported by Suda et al.
(2000) and Neves et al. (2001). Cooking significantly (P ≤ 0.05)
reduced globulin they were 16.4%, 19.0%, 18.1%, 18.0%, 17.8% and
17.8% for Ain Elgazal raw the first, second, third and fourth days
respectively. Similarly for Buff they were 15.7%, 19.9%, 22.0%,
22.6% and 23.0% respectively, a result which are simlar to that
reported by Nugdallah and El Tinay (1997); Fiel et al. (2003).
110
Autoclaving at 120ºC under 15 psi for 30 minutes and at 150ºC under
20 psi for 30 minutes for Ain Elgazal raw seeds significantly (P ≤
0.05) reduced globulin fraction from 87.5% - 29.2% and 27.6%
respectively. Similarly for Buff raw seeds autoclaving at 120ºC under
15 psi for 30 minutes and at 150ºC under 20 psi for 30 minutes
significantly (P ≤ 0.05) reduced globulin fraction from 89.8% - 32.0%
and 36.0% respectively. Dry heating as well as cooking and
autoclaving significantly (P ≤ 0.5) reduced globulin fraction.
Roasting Ain Elgazal raw seeds at 90ºC/60 min and 120ºC/60
min significantly (P ≤ 0.05) decreased globulin fraction from 87.5% to
85.0% and 41.0% respectively. Similarly for Buff raw seeds roasing at
90ºC/60 min and 120ºC/60 min significantly (P ≤ 0.05) decreased
globulin fraction from 89.8% to 88.0% and 45.5% respectively, a
result which are similar to that reported by Nugdallah and El Tinay
(1997); Fiel (2003).
4.2.2. Albumin fraction:
cooked, autoclaved and roasted seeds were fractionated. The albumin
fraction significantly (P ≤ 0.05) decreased, they were 4.0%, 3.8%,
2.3%, 1.9%, and 2.5% for Ain Elgazal raw, the first, second, third, and
fourth day respectively. Similarly for Buff they were 3.6%, 1.5%,
111
2.2%, 2.2% and 1.2% respectively, a result which are similar to that
reported by Nugallah and El Tinay (1997), Sud et al. (2000); Neves
et al. (2001), but were different from those reported by Giami et al.
(1999); and El Khalifa and El Tinay (1999). Cooking significantly (P
≤ 0.05) reduced albumin, they were 1.9%, 2.8%, 3.1%, 1.8%, and
1.4% for Ain Elgazal raw the first, second, third and fourth days
respectively. Similarly for Buff they were 2.2%, 1.6%, 1.6%, 2.1%
and 1.3% respectively. Autoclaving at 120ºC under 20 psi for 30
minutes for Ain Elgazal raw seeds significantly (P ≤ 0.05) reduced
albumin fraction from 4.0%-1.2% and 1.2% respectively. Similarly for
Buff raw seeds autoclaving at 120ºC under 15 psi for 30 minutes and
at 150ºC under 20 psi for 30 minutes significantly (P ≤ 0.05) reduced
globulin fraction from 3.6% - 1.0% and 0.9% respectively. Roasting
as well as cooking and autoclaving significantly (P ≤ 0.5) reduced
albumin fraction. Roasting Ain Elgazal raw seeds at 90ºC/30 min
albumin in fraction showed changeless, but roasting at 120ºC/60 min
significantly (P ≤ 0.05) reduced albumin fraction from 4.0% to 1.2%.
Similarly for Buff raw seeds roasing at 90ºC/30 min albumin fraction
showing changeless, but roasting at 120ºC/60 min significantly (P ≤
0.05) reduced albumin fraction from 3.6% to 1.0%, a result which are
112
similar to that reported by Nugdallah and El Tinay (1997); Fiel
(2003).
4.2.3. Prolamin fraction:
cooked, autoclaved and roasted seeds were fractionated. The prolamin
fraction significantly (P ≤ 0.05) decreased, they were 4.0%, 3.3%,
4.0%, 1.6%, respectively. Similarly for Buff raw and germinated seeds
they were 4.5%, 2.7%, 2.8%, 3.9% and 1.4% respectively, a result
which are similar to that reported by Giami et al. (1999); Khalifa and
El Tinay (1999). Cooking significantly (P ≤ 0.05) increased prolamin
fraction, they were 4.3%, 6.3%, 4.8%, 5.1%, and 5.0% for Ain Elgazal
raw the first, second, third and fourth days respectively. Similarly for
Buff they were 4.6%, 5.1%, 2.7%, 2.7% and 1.8% respectively. It
showed significant (P ≤ 0.05) increase during the first day of
germination but started to decrease significantly (P ≤ 0.05), reaching a
minimum value after 96h of germination, a result which are similar to
that reported by Nugdallah and El Tinay (1997). Autoclaving at 120ºC
under 15 psi for 30 minutes and at 150ºC under 20 psi for 30 minutes
for Ain Elgazal raw seeds significantly (P ≤ 0.05) increased prolamin
fraction from 4.3% - 4.5% and 4.8% respectively. For Buff raw seeds
autoclaving at 120ºC under 15 psi for 30 minutes and at 150ºC under
20 psi for 30 minutes, significantly (P ≤ 0.05) decreased prolamin
fraction from 4.5% - 3.0% and 4.0% respectively. Roasting as well as
cooking and autoclaving significantly (P ≤ 0.5) decreased prolamin.
Roasting Ain Elgazal raw seeds at 90ºC/30 min and at 120ºC/60 min
significantly (P ≤ 0.05) reduced prolamin fraction from 4.3% to 3.6%,
and 4.2%. For Buff raw seeds significantly (P ≤ 0.05) reduced
113
prolamin from 4.5% to 4.0% but roasting at 120ºC/60min significantly
(P ≤ 0.05) increased prolamin fraction from 4.5% to 6.0% respectively.
A result which are similar to that reported by Nugdallah and El Tinay
(1997).
4.2.4. G1-glutelin:
As shown in Tables 3, 4 and 5, the G1- glutelin fraction for Ain
Elgazal raw and germinated seeds significantly (P ≤ 0.05) decreased,
they were 2.3%, 1.5%, 1.3%, 0.8%, and 1.7%, respectively. For Buff
raw and germinated seeds they were 2.3%, 1.2%, 1.3%, 2.1% and
1.3% respectively, a result which are similar to that reported by El
Khalifa et al., (1996). Cooking significantly (P ≤ 0.05) decreased G1glutelin fraction, they were 2.0%, 1.4%, 1.3%, 1.3%, and 2.6% for Ain
Elgazal raw the first, second, third and fourth days respectively, but
started to increase 96h of germinnat. For Buff raw and germinated
seeds, it showed significant (P ≤ 0.05) decreased they were 2.7%,
1.2%, 1.1%, 1.6% and 1.0% respectively, a result which are similar to
that reported by Nugdallah and El Tinay (1997).
Autoclaving at 120ºC under 15 psi for 30 minutes and at 150ºC
under 20 psi for 30 minutes for Ain Elgazal raw seeds significantly (P
≤ 0.05) decreased G1-glutelin fraction from 2.3% - 1.2% and 2.0%
respectively. For Buff raw seeds autoclaving significantly (P ≤ 0.05)
decreased G1-glutelin from 2.3% to 2.0% and 1.2% respectively.
114
Roasting as well as cooking and autoclaving significantly (P ≤ 0.5)
decreased G1-glutelinfor Ain Elgazal raw seeds from 2.3% to 2.0%
and 1.2% respectively. For Buff raw seeds G1-glutelin significantly (P
≤ 0.05) decreased from 2.3% to 2.2% and 2.0% respectively, a result
which are similar to that reported by Nugdallah and El Tinay (1997).
4.2.5. G2-glutelin:
As shown in Tables 3, 4 and 5, the G2- glutelin fraction
for Ain Elgazal raw and germinated seeds significantly (P ≤ 0.05)
decreased, they were 2.4%, 2.1%, 1.9%, 1.4% and 1.7%, respectively,
a result which are similar to that reported by El Khalifa et al. (1996).
For Buff raw and germinated seeds G2-glutelin showed a
significantly (P ≤ 0.05) increased, they were 2.5%, 6.4%, 5.1%, 3.9%
and 4.5% respectively, a result which are similar to that reported by
El Khalifa et al., (1996). Cooking significantly (P ≤ 0.05) decreased
G1-glutelin fraction, they were 3.4%, 4.4%, 4.4%, 4.0%, and 5.7% for
Ain Elgazal raw the first, second, third and fourth days respectively.
Similarly for Buff raw and germinated seeds, they were 5.1%, 7.5%,
7.5%, 7.9% and 8.8% respectively, a result which are similar to that
reported by Nugdallah and El Tinay (1997); Fiel et al. (2003).
Autoclaving at 120ºC under 15 psi for 30 minutes significantly
(P ≤ 0.05) decrease G2-glutelin from 2.4% to 1.2% and 4.0%
115
autoclaving at 150ºC under 20 psi for 30 minutes significantly (P ≤
0.05) increased from 2.5% to 3.0% and 3.2% respectively. Roasting
Ain Elgazal raw seeds at 90ºC for 60 minutes significantly (P ≤ 0.05)
decreased G2-glutelin fraction from 2.4% - 1.2% and roasting the
seeds at 120ºC for 60 minutes significantly (P ≤ 0.05) increased G2glutelin from 2.4% to 3.2%. Similarly for Buff raw seeds roasting at
90ºC for 60 minutes and 120ºC for 60 minutes significantly (P ≤ 0.05)
increased G2-glutelin from 2.5% to 2.8% and 3.0% respectively, a
(1997); Fiel et al. (2003).
4.2.6. G3-glutelin:
As shown in Tables 3, 4 and 5, the G3- glutelin fraction
significantly (P ≤ 0.05) increased, they were 4.8%, 9.8%, 12.0%,
13.5% and 10.4%, for Ain Elgazal raw seeds, the first, second, third
and fourth days respectively, a result which are similar to that reported
by El Khalifa et al. (1996).
For Buff raw and germinated seeds G2-glutelin showed a
significantly (P ≤ 0.05) increased, they were 4.5%, 10.0%, 12.5%,
11.1% and 11.7% respectively, a result which are similar to that
reported by El Khalifa et al. (1996). Cooking significantly (P ≤ 0.05)
decreased G3-glutelin fraction, they were 63.8%, 61.0%, 62.0%,
116
61.6%, and 57.0% for Ain Elgazal raw the first, second, third and
fourth days respectively. Similarly for Buff raw and germinated seeds,
they were 69.3%, 59.0%, 62.3%, 60.1% and 62.1% respectively, a
(1997); Fiel et al. (2003). Autoclaving at 120ºC under 15 psi for 30
minutes and at 150ºC under 20 psi for 30 minutes significantly (P ≤
0.05) increased G3-glutelin for Ain Elgazal raw seeds from 4.8% to
59.5% and 60.2% respectively. For Buff seeds G3-glutelin
significantly (P ≤ 0.05) increased from 4.5% to 55.0% and 50.8%
respectively.
Roasting Ain Elgazal raw seeds at 90ºC for 60 minutes and at
120ºC for 60 minutes significantly (P ≤ 0.05) increased G3-glutelin
from 4.8% - 6.0% and 48.0% respectively, for Buff raw seeds G3glutelin showed changeless at 90ºC for 60 minutes but at 120ºC for 60
minutes showed significantly (P ≤ 0.05) increased they were 4.5% and
42.0% respectively, a result which are similar to that reported by
Nugdallah and El Tinay (1997); Fiel et al. (2003).
4.2.7. Residue fraction:
As shown in Tables 3, 4 and 5, residue fraction showed
significant (P ≤ 0.05) increased, they were 1.2%, 2.5%, 2.1%, 9.2%
and 6.0%, for Ain Elgazal raw seeds, the first, second, third and fourth
117
days respectively, similarly for Buff raw and germinated seeds, they
were 1.2%, 1.7%, 1.9%, 3.6% and 6.1% respectively, a result which
are similar to that reported by El Khalifa and El Tinay (1999).
Cooking significantly (P ≤ 0.05) decreased G3-glutelin fraction, they
were 5.5%, 8.0%, 8.8%, 9.6%, and 10.2% for Ain Elgazal raw the
first, second, third and fourth days respectively. Similarly for Buff raw
and germinated seeds, they were 5.5%, 6.6%, 6.5%, 6.1% and 6.5%
respectively, a result which are similar to that reported by Nugdallah
and El Tinay (1997); Fiel et al. (2003).
Autoclaving at 120ºC under 15 psi for 30 minutes and at 150ºC
under 20 psi for 30 minutes significantly (P ≤ 0.05) increased fractions
for Ain Elgazal raw seeds from 1.2% to 8.0% and 6.0% respectively.
For Buff seeds residue fractions significantly (P ≤ 0.05) increased from
1.2% to 6.0% and 7.0% respectively. Roasting Ain Elgazal raw seeds
at 90ºC for 60 minutes and at 120ºC for 60 minutes significantly (P ≤
0.05) increased residue fraction from 1.2% - 3.0% and 6.0%
respectively.
For Buff raw seeds residue fraction showed significant (P ≤
0.05) increased at 90ºC for 60 minutes but at 120ºC for 60 minutes
from 1.2% to 1.4% and 6.0% respectively, a result which are similar
to that reported by Nugdallah and El Tinay (1997); Fiel et al. (2003).
118
Table 3. Effect of germination on protein fractions of cowpea cultivars
Cultivar
Ain-Elgazal
Buff
G3-glutelin
Residue
%
%
2.4±0.00a
4.8±0.07e
1.2±0.00d
106.5
1.5±0.03b
2.1±0.01b
9.8±0.01d
2.5±0.00c
103.6
4.0±0.00b
1.3±0.01c
1.9±0.01c
12.0±0.02b
2.1±0.01c
100.0
1.9±0.03c
1.6±0.00d
0.8±0.00d
1.4±0.02e
13.5±0.1a
9.2±0.01a
99.8
70.2±0.61e
2.5±0.01b
4.6±0.1a
1.7±0.00b
1.7±0.00d
10.4±0.04c
6.0±0.00b
97.1
Raw seeds
89.8±0.07a
3.6±0.01b
4.5±0.12d
2.3±0.03a
2.5±0.00e
4.5±0.04e
1.2±0.00d
108.4
24
81.3±0.13b
1.5±0.00c
2.7±0.21d
1.2±0.00d
6.4±0.04a
10.0±0.03d
1.7±0.00c
104.8
48
78.3±0.04c
2.2±0.00c
2.8±0.00c
1.3±0.07c
5.1±0.03b
12.5±0.04a
1.9±0.01c
104.1
72
75.6±0.02d
2.2±0.00b
3.9±0.00b
2.1±0.00b
3.9±0.04d
11.1±0.12c
3.6±0.01b
102.4
96
72.7±0.03e
1.2±0.01d
1.4±0.00e
1.3±0.13c
4.5±0.00c
11.7±0.00b
6.1±0.01a
98.9
Germination
Globulin
Albumin
Prolamin
time, h
%
%
%
Raw seeds
87.5±0.02a
4.0±0.15a
24
80.6±0.91b
48
G1-glutelin %
G2-glutelin %
4.3±0.25a
2.3±0.09a
3.8±0.06a
3.3±0.03c
76.4±0.03c
2.3±0.03b
72
71.4±0.04d
96
Total proteins
Values are means (±SD),
assessed by Duncan's Multiple Range Test
119
Table 4. Effect of cooking on protein fractions of germinated cowpea cultivars
Cultivar
Ain-Elgazal
Buff
G3-glutelin
Residue
Total proteins
%
%
%
3.4±0.04d
63.8±0.04a
5.5±0.01e
97.2
1.4±0.00c
4.4±0.04b
61.0±0.04b
8.0±0.00d
102.9
4.8±0.16c
1.3±0.00c
4.4±0.04b
62.0±0.07b
8.8±0.04c
102.5
1.8±0.00c
5.1±0.00bc
1.3±0.09c
4.0±0.03c
61.6±0.04b
9.6±0.00b
101.4
17.8±0.04d
1.4±0.01d
5.0±0.16c
2.6±0.00a
5.7±0.41a
57.0±0.00c
10.2±0.01a
99.7
0
15.7±0.02e
2.2±0.00a
4.5±0.00b
2.7±0.02a
5.1±0.07d
69.3±0.04a
5.5±0.01d
105.1
24
19.9±0.04d
1.6±0.02c
5.1±0.02ab
1.2±0.00c
7.5±0.04c
59.0±0.04cd
6.6±0.01a
100.9
48
22.0±0.06c
1.6±0.01c
2.7±0.03c
1.1±0.09c
7.5±0.00c
62.3±0.04b
6.5±0.01b
103.7
72
22.6±0.04b
2.1±0.01ab
2.7±0.00c
1.6±0.00b
7.9±0.02bc
60.1±0.04c
6.1±0.01c
103.1
96
23.0±0.00a
1.3±0.01d
1.8±0.07d
1.0±0.00d
8.8±0.04
62.1±0.03b
6.5±0.01b
104.7
Germination
Globulin
Albumin
Prolamin
time, h
%
%
%
0
16.4±0.00e
1.9±0.03c
24
19.0±0.04a
48
G1-glutelin %
G2-glutelin %
4.3±0.04d
2.0±0.00b
2.8±0.04b
6.3±0.00a
18.1±0.04b
3.1±0.01a
72
18.0±0.04bc
96
120
Table 5. Effect of autoclaving and roasting on protein fractions of cowpea cultivars
Treatment
Globulin
%
Albumin
%
Prolamin
%
G1-glutelin
%
G2-glutelin
%
G3-glutelin
%
Residue
%
Total P.
recovered %
87.5±0.02a
29.2±0.4d
27.6±0.2e
4.0±0.015a
1.2±0.1b
1.2±0.1b
4.3±0.25c
4.5±0.1b
4.8±0.1a
2.3±0.09a
1.2±0.0c
2.0±0.0b
2.4±0.0c
1.2±0.4d
4.0±0.0a
4.8±0.07cd
59.5±0.1a
60.2±0.1a
1.2±0.00d
8.0±0.3a
6.0±0.1b
106.5
104.8
105.8
85.0±0.3b
41.0±0.4c
4.0±0.2a
1.2±0.3b
3.6±0.2e
4.2±0.4d
2.0±0.1b
1.2±0.6c
1.2±0.0d
3.2±1.3b
6.0±0.2c
48.0±0.3b
3.0±0.0c
6.0±0.4b
104.8
104.8
89.8±0.07a
32±0.1e
36±0.2d
3.6±0.01a
1.0±0.01b
0.9±0.02c
4.5±0.12b
3.0±0.01d
4.0±0.01c
2.3±0.03a
2.0±0.1c
1.2±0.09d
2.5±0.00d
3.0±0.03b
3.2±0.02a
4.5±0.04d
55.0±0.02a
50.8±0.1b
1.2±0.00d
6.0±0.01b
7.0±0.1a
108.4
102.0
103.1
88.0±0.3b
45.5±1.3c
3.6±0.2a
1.0±0.01b
4.0±0.02c
6.0±0.1a
2.2±0.01b
2.0±0.1c
2.8±0.02c
3.0±0.00b
4.5±0.01d
42.0±0.06c
1.4±0.2c
6.0±0.00b
106.5
105.5
Ain Elgazal
Raw seeds
Autoclave
15 lb/30 min
20 lb/30 min
Roasting
90ºC/60 min.
120ºC/60 min.
Buff
Raw seeds
Autoclave
15 lb/30 min
20 lb/30 min
Roasting
90ºC/60 min.
120ºC/60 min.
Values are means (±SD)
121
4.3. anti-nutrient:
4.3.1. Tannins:
As shown in Tables 6 germination significantly (P ≤ 0.05)
reduced Tannins from 0.48 g/100g of raw seeds to 0.36g/100g,
0.24g/100g and 0.20g/100g for Ain Elgazal germinated seeds, the
first, second, third and fourth days respectively. Similarly for Buff raw
seeds, Tannins was 0.50g/100g which significantly (P ≤ 0.05) reduced
by germination to 0.42g/100g, 0.36g/100g, 0.30g/100g and 0.22g/100g
for the first, second, third and fourth days respectively. Cooking
significantly (P ≤ 0.05) reduced Tannins for Ain Elgazal raw and
germinated seeds, they were 0.26g/100g, 0.18g/100g, 0.15g/100g
0.12g/100g and 0.10g/100g respectively. Similarly for Buff, cooking
significantly (P ≤ 0.05) reduced tannins, they were 0.30g/100g,
0.22g/100g, 0.20g/100g, 0.18g/100g and 0.11g/100g respectively.
Autoclaving significantly (P ≤ 0.05) decreased Tannins for Ain
Elgazal raw seeds 0.48g/100g, 0.38g/100g, Similarly for Buff raw
seedsautoclaving at 150ºC under 20 psi for 30 minutes significantly (P
≤ 0.05) decreased Tannins from 0.50g/100g to 0.40g/100g. Roasting
Ain Elgazal raw seeds significantly (P ≤ 0.05) reduced Tannins from
0.48g/100g to 0.36g/100g. Similarly for Buff raw seeds roasting
significantly (P ≤ 0.05) decreased Tannins from 0.50g/100g to
122
0.42g/100g, a result which are similar to that reported by Anthony et
al. (1984); Ene-Obong (1995); Ningsanond et al. (1989); Ekpenyong
(1985); Bakr et al. (1991); Chen et al. (1977); Plahar (1997); Vaishale
et al. (1998); Jibaja et al., (1988); Laurena et al. (1987); Mugula et
al., (1999); Vijayakumari et al. (1998). Results obtained were
different from those reported by Oke et al., (1996) and Beem Van et
al., (1992).
4.3.2. Trypsin inhibitor activity (TIA):
reduced Trypsin inhibitor activity for Ain Elgazal raw and germinated
seeds, they were 22.0 TUI/mg protein, 11.8 TUI/mg protein, 10.6
TUI/mg protein, 8.0 TUI/mg protein and 8.0 TUI/mg protein,
respectively. Similarly for Buff raw and germinated seeds TIA were
25.0 TUI/mg protein, 12.5 TUI/mg protein, 10.0 TUI/mg protein, 9.6
TUI/mg protein and 9.0 TUI/mg protein, respectively. Cooking
autoclaving and raosting eliminates TIA, a result which are similar to
that reported by Anthony et al. (1984); Carnovale et al., (1992)
Beltran et al., (1983); Iss et al., (1996); Mulimani et al., (1994).
4.3.3. Phytic acid:
reduced Phytic acid from 310.3 (mg/100g dry weight) of raw seeds to
123
286.1 mg/100g, 248.9 mg/100g, 201.7 mg/100g, and 139.8 mg/100g,
for Ain Elgazal germinated seeds, the first, second, third and fourth
days respectively. Similarly for Buff raw seeds, phytic acid were
376.3 mg/100mg which significantly (P ≤ 0.05) reduced by
germination to 346.2 mg/100g,, 301.0 mg/100g, 225.7 mg/100g, and
180.7 mg/100g, for the first, second, third and fourth days
respectively. Cooking significantly (P ≤ 0.05) they decreased Phytic
acid for Ain Elgazal raw and germinated seeds, they were 290.3
mg/100g, 268.9 mg/100g, 229.0 mg/100g, 181.5 mg/100g, and 128.0
mg/100g, respectively. Similarly for Buff cooking significantly (P ≤
0.05) decreased Phytic acid they were, 353.0 mg/100g, 311.6
mg/100g, 270.9 mg/100g, 205.4 mg/100g, and 162.6 mg/100g,
respectively. Autoclaving significantly (P ≤ 0.05) decreased Phytic
acid for Ain Elgazal raw seeds from 310.3 mg/100g, to 300.0
mg/100g,. similrly for Buff raw seeds autoclaving at 150ºC under 20
psi for 30 minutes significantly (P ≤ 0.05) decreased Phytic acid from
376.3 mg/100g, to 350.0 mg/100g. Roasting at 120ºC for 60 min
significantly (P ≤ 0.05) decreased Phytic acid for Ain Elgazal raw
seeds from 310.3 mg/100g, to 300.1 mg/100g. Similarly for Buff raw
seeds roasting significantly (P ≤ 0.05) decreased Phytic acid from
376.3 mg/100g, to 352.0 mg/100g, a result which are similar to that
124
reported by Anthony et al. (1984); Alonso et al., (1998); Chitra
(1996); Anshu et al., (1995); Sanni et al., (1999); Perumal-Siddhuraju
et al., (1996); Ene – obong et al., (1996); Yadav et al., (1994); Ogum
et al., (1989); Uzogara et al., (1997) Ologhobo et al., (1984); Chem
et al., (1997); Vanderstop (1981); Baralker et al., (1985);
Alxiny
et al., (1991); Ene – Obong (1995); Vijayakumari et al., (1998).
Bakr et al., (1991), results obtained were different from those
reportedb by Oke et al., (1996).
4.4. In-vitro protein digestibilty (IVPD):
4.4.1. Effect of processing on IVPD:
Improvement of protein digestibility after processing could be
attributable to the reduction or elimination of different antinutrients.
Phytic acid, as well as condenced tannins and polyphenols which are
known to interact with protein to form complexes. This interaction
increases the degree of cross-linking, decreasing the solubility of
proteins and making protein complexes less succeptable to proteolyic
attack than the same protein alone, Alonso et al., (2000). As shown in
table 6 germination significantly (P ≤ 0.05) improved in-vitro protein
digestibility (IVPD), they were 73.4%, 75.3%, 77.9%, 80.4%, and
84.4% for Ain Elgazal raw seeds, the first, second, third, and fourth
days respectively. Similarly for Buff, they were 74.2%, 75.7%, 79.3%,
125
82.4% and 83.6% respectively. Cooking germinated seeds showed
significantly (P ≤ 0.05) further increase of in-vitro protein digestibility,
they were 86.2%, 87.2%, 87.5%, 88.8%, and 88.5% for Ain Elgazl
raw seeds, the first, second, third, and fourth days respectively.
Similarly for Buff, they were 85.4%, 86.3%, 86.6%, 87.9% and 88.3%
for raw seeds, the first, second, third, and fourth days respectively.
Autoclaving at 150ºC under 20 psi for 30 min significantly (P ≤
0.05) increased in vitro protein digestibilty for Ain Elgazal raw seeds
from 73.4%, to 86.0% and significantly (P ≤ 0.05) increased by
roasting at 120ºC for 60 min from 73.4% to 84.0%. Similrly for Buff
raw seeds IVPD significantly (P ≤ 0.05) increased by autoclaving
from 74.2% to 87.0% and by roasting from 74.2% to 84.0%
respectively, a result which are similar to that reported by Anthony
et al. (1988); Nnanno and Phillips (1989); Jibaja et al., (1988).
Laurena et al., (1987); Mugula et al., (1996); Chitra (1996); PerumalSiddhuraju et al., (1996); Ene-obong et al., (1996); Vijayakumari et
al., (1998); Anshu-Sharma et al., (1995) Yadav et al., (1994);
Vanderstop (1981); Baralker et al., (1985).
126
Table 6. Phytic acid (mg/100g dry weight), Tannins (g/100g dry weight), trypsin inhibitor activity (TUI/mg protein)
and IVPD of raw and processed cowpea cultivar
Treatment
Ain Elgazal
Raw seeds
Germinated 24 h.
Germinated 48 h.
Germinated 72 h.
Germinated 96 h.
Autoclaved 20 lb/30 min.
Roasting 120ºC/60 min.
Buff
Raw seeds
Germinated 24 h.
Germinated 48 h.
Germinated 72 h.
Germinated 96 h.
Autoclaved 20 lb/30 min.
Roasting 120ºC/60 min.
Tannic acid
Uncooked Cooked
TIA
Uncooked
Cooked
Phytic acid
Uncooked Cooked
IVPD
Uncooked
Cooked
0.48±0.2
0.36±0.02
0.30±0.2
0.24±0.1
0.20±0.02
-
0.26±0.01
0.18±0.5
0.15±0.3
0.12±0.3
0.10±0.01
0.38±0.03
0.36±0.1
22.0±0.5
11.8±0.01
10.6±0.01
8.0±0.1
8.0±0.1
-
Nil
-
310.3±1.3a
286.1±0.6b
248.9±1.4c
201.7±1.3d
139.8±1.4e
-
290.3±1.4a
268.9±1.3b
229.0±1.9c
181.5±1.8d
128.6±0.4e
300.0±0.1
301.0±0.3
73.4±1.33e
75.3±1.44d
77.9±0.4c
80.4±0.4b
84.4±0.3a
-
86.2±0.6bc
87.2±0.5b
87.5±0.5b
88.8±0.4a
88.5±0.2a
86.0±c
84.0±d
0.50±0.01
0.42±0.01
0.36±0.01
0.30±0.02
0.22±0.01
-
0.30±0.02
0.22±0.01
0.20±0.02
0.18±0.01
0.11±0.01
40.0±0.1
42.0±0.2
25.0±0.1
12.5±0.1
10.0±0.01
9.6±0.01
9.0±0.2
-
Nil
-
376.3±0.4a
346.2±01b
301.0±0.2c
125.7±0.1d
180.7±0.3e
-
353.7±0.6a
311.6±0.1b
270.9±02c
205.4±0.1d
162.6±0.5e
350±0.1
352±0.1
742±0.5e
75.7±0.6d
79.3±1.86c
82.4±0.4b
83.6±0.3a
-
85.4±0.6cd
86.3±0.5a
86.6±0.4ab
87.9±0.4
88.3±0.4c
87.0±0.1b
84.0±0.1e
127
4.4.2. Effect of substract on in-vitro protein digestibilty:
Effect of substrate on in-vitro protein digestibilty was carried
out using mixed proteins of raw with treated sample in various
proteins.
Increasing the protein of treated in the comparison to raw
material as shown in Table 7 significantly (P ≤ 0.05) increased in-vitro
protein digestibilty for both cultivars. The rate of increase for the
proportional for amount of treated samples. This could attributed to
heat inactivation to anti-nutritional factors which are expected to be of
higher concentration in raw material. Ain Elgazal raw proportions
were 0.384g, 0.336g, 0.288g, 0.00g, 0.048g, 0.096g and 0.192g which
were mixed with roasted samples, the proportions of roasted samples
were 0.00g, 0.048g, 0.096g, 0.384g, 0.0336g, 0.0282g, and 0.912g the
sequences were A0, A1, A2, A3, A4, A5 and A6 respectively. Raw plus
autoclaved samples sequences were A7, A8, A9, A10, A11, A12 and A13
respectively. Roasting was at 90ºC and at 120ºC for 30, 45 and 60
min, autoclaving was at 115.5ºC under 10 psi and at 120ºC under 15
psi for 15, 30, and 45 min. In-vitro protein digestibilty for the above
sequence were 73.4%, for A0, they were 73.4%, 73.4%, 73.5%,
73.5%, 73.5% 73.6% and 73.8% respectively for A1, they were 73.5%,
73.6%, 73.6%, 73.9%, 73.8% and 74.0% respectively for A2, they
128
were 80.0%, 80.1%, 80.2%, 83.9%, 84.0%, and 84.2% respectively
for A3, they were 79.5%, 79.6%, 79.8%, 83.0%, 83.2%, and 83.1%
respectively for A4, they were 75.0%, 75.3%, 75.2%, 75.4%, 75.5%,
and 76.0% respectively for A5, they were 73.8%, 73.8%, 74.0%,
74.8%, 75.1%, and 75.0% respectively for A6, they were 73.4% for
A7, they were 73.8%, 74.0%, 74.2%, 74.1%, 74.0%, and 74.6%
respectively for A8, they were 74.9%, 74.8%, 74.8%, 74.2%, 75.0%,
and 75.8% respectively for A9, they were 82.0%, 83.0%, 84.0%,
85.0%, 85.5% and 86.2% respectively for A10, they were 81.0%,
81.8%, 82.8%, 83.5%, 83.3%, and 84.0% respectively for A11, they
for A12, they were 74.8%, 74.8%, 74.9%, 74.8%, 75.2%, and 75.0%
respectively for A13,
Similarly for Buff cultivar the raw proportion were 0.40g,
0.35g, 0.30g, 0.00g, 0.05g, 0.10 and 0.20g which were mixed with
roasted samples, the proportion of roasted sample were 0.00g, 0.05g,
0.10g, 0.40g, 0.35g, 0.30g and 0.20g in sequences which were B0, B1,
B2, B3, B4, B5 and B6 respectively. Raw plus autoclaved samples
sequences were B7, B8, B9, B10, B11, B12 and B13 respectively. In-vitro
protein digestibilty for the above sequence were 74.2%, for B0, they
were 74.5%, 74.6%, 74.6%, 75.5%, 75.6% and 75.0% respectively for
129
B1, they were 75.0%, 75.1%, 75.0%, 76.0%, 76.0 and 75.3%
respectively for B2, they were 79.7%, 79.9%, 79.8%, 83.8%, 84.0%,
and 84.8% respectively for B3, they were 78.7%, 78.8%, 78.8%,
82.3%, 82.9%, and 83.0% respectively for B4, they were 76.0%,
76.0%, 76.0%, 76.8%, 76.8%, and 77.2% respectively for B5, they
for B6, they were 74.2% for B7, they were 74.5%, 74.8%, 74.9%,
75.1%, 75.0%, and 75.0% respectively for B8, they were 75.7%,
75.6%, 75.4%, 75.8%, 76.0% and 76.4% respectively for B9, they
for B10, they were 79.0%, 82.0%, 83.0%, 83.0%, 84.0%, and 84.0%
respectively for B11, they were 76.0%, 76.5%, 76.8%, 77.0%, 77.1%,
and 77.3% respectively for B12, they were 75.0%, 75.0%, 75.0%,
75.5%, 75.6%, and 76.0% respectively for B13, a result which are
similar to that reported by Plahar (1997); Vaishale et al., (1998);
Jibaja et al., (1988); Laurena et al., (1987); Yadav et al., (1994);
Savage et al., (1993); Mugula et al., (1999).
130
Table 7. Effect of Substrate on IVPD:
In ivtro protein digestibility of mixed sample
weight
Roasted sample
Autoclaved sample
30min
45min
60min
30min
45min
60min
Sequences
Buff
Ain Elgazal
Sequences
Sample
15min
30min
45min
15min
30min
45min
A0
0.384±0.0
73.4±1.3cd
73.4±1.3cd
73.4±1.3cd
73.4±1.3de
73.4±1.3de
73.4±1.3d
A7
73.4±1.3d
73.4±1.3de
73.4±1.3de
73.4±1.3de
73.4±1.3de
73.4±1.3de
A1
0.336±0.048
73.4±0.3cd
73.4±0.6cd
73.5±0.4cd
73.5±0.4de
73.6±0.4cde
73.8±0.4cd
A8
73.8±0.6d
74.0±0.6d
74.2±0.9de
74.1±0.3d
74.0±0.4d
74.6±0.06de
A2
0.288±0.096
73.5±0.5c
73.6±0.5c
73.6±0.4c
73.9±0.3de
73.8±0.6cd
74.0±0.5cd
A9
74.9±1.3cd
74.8±0.5cd
74.8±0.4cd
74.2±0.6d
75.0±0.3cd
75.8±0.6cd
A3
0.0±0.384
80.0±0.3a
80.1±0.5a
80.2±0.6a
83.9±0.6a
84.0±0.6a
84.2±0.4a
A10
82.0±0.6a
83.0±0.5a
84.0±0.6a
85.0±0.4a
85.5±0.6a
86.2±0.4a
A4
0.048±0.336
79.5±0.4a
79.6±0.4a
79.8±0.5a
83.0±1.3ab
83.2±1.3ab
83.1±0.3ab
A11
81.0±0.5b
81.8±0.5ab
82.8±0.4ab
83.5±0.4b
83.3±0.5b
84.0±0.5b
A5
0.096±0.282
75.0±1.3b
75.3±0.5b
75.2±0.4b
75.4±0.6c
75.5±0.5c
76.0±0.4c
A12
75.5±0.5c
75.6±0.5c
75.7±0.6c
75.7±0.5c
76.2±0.4c
76.3±0.4c
A6
0.192±0.192
73.8±0.4
73.8±0.3c
74.0±1.3bc
74.8±0.4d
75.1±0.6cd
75.0±1.3cd
A13
74.8±0.4cd
74.8±0.4cd
74.9±0.6cd
74.8±0.4d
75.2±0.5cd
75.0±0.4cd
B0
0.4±0.0
74.2±0.4d
74.2±0.4de
74.2±0.4e
74.2±0.4e
74.2±0.4ed
74.2±0.4cd
B7
74.2±0.4de
74.2±0.4de
74.2±0.4e
74.2±0.4e
74.2±0.4e
74.2±0.2cd
B1
0.3±0.1
74.5±0.4d
74.6±0.4d
74.6±0.6d
75.5±0.5d
75.6±0.4cd
75.0±0.4d
B8
74.5±0.4d
74.8±0.3de
74.9±0.3de
75.1±0.3de
75.0±0.5de
75.0±0.4bc
B2
0.35±0.05
75.0±0.4d
75.1±0.4d
75.0±0.4cd
76.0±0.4cd
76.0±0.3d
75.3±0.3d
B9
75.7±0.4c
75.6±0.3d
75.4±0.9d
75.8±1.3d
76.0±0.0d
76.4±0.4b
B3
0.0±0.4
79.7±0.3a
79.9±0.4a
79.8±0.5a
83.8±0.6a
84.0±0.5a
84.8±0.3a
B10
80.2±0.5a
84.8±0.4a
84.8±0.6a
85.0±0.4a
85.0±0.4a
85.4±0.5a
B4
0.05±0.35
78.7±0.4b
78.8±0.3b
78.8±0.3b
82.3±0.5b
82.9±1.3b
83.0±0.3b
B11
79.0±0.6b
82.0±0.4b
83.0±0.6b
83.0±0.4b
84.0±0.5ab
84.0±0.4a
B5
0.10±0.3
76.0±0.4c
76.0±0.4c
76.0±0.4c
76.8±0.4c
76.8±0.6c
77.2±0.6c
B12
76.0±0.5c
76.5±0.3c
76.8±0.5c
77.0±0.4c
77.1±0.6c
77.3±1.3b
B6
0.2±0.2
74.6±1.3d
74.5±1.3de
74.5±1.3de
75.1±0.4de
75.2±0.6cd
75.0±0.5d
B13
75.0±0.4cd
75.0±0.6d
75.0±0.4d
75.5±0.3d
75.6±1.3d
76.0±0.5b
Raw (ing)
Treated (ing)
90ºC
120ºC
115..5ºC 10psi
120.5ºC 15spi
assessed by Duncan's Multiple – Range Test.
131
CHAPTER FIVE
SUMMARY, CONCLUSION AND
RECOMMENDATION
132
CHAPTER FIVE
SUMMARY AND CONCLUSIONS
Two cowpea cultivars were analyzed for proximate composition. Treatment
included cooking, roasting, autoclaving, germinating and cooking plus germinated
seeds. The proteins of control and treated sample were fractionated according to
solubility behaviour. The results indicated that the albumin plus globulin fractions
decreased significantly (P ≤ 0.05) for all treated sample, however, there was high
retention of globulin fraction in autoclaved sample compared to ordinary cooked
ones. For all treated samples decrease in the albumin and globlin fraction was a
companied by significant (P ≤ 0.05) increase in G3-glutelin fraction, the IVPD on
processed cowpeas was significantly (P ≤ 0.05) improved. This was more
pronounced for cooked germinated and autoclaved samples. This was associated with
significant (P ≤ 0.05) reduction of the anti-nutritional factor for all treatment.
133
RECOMMENDATION:
The acute shortage of meat and animal proteins in developing countries has
made it necessary for consumer in these countries to rely heavily on proteins from
legumes spically cowpeas. These beans, which are rich in protiens, B-vitamins and
dietary fiber, are important in the diet of people in developing countries where
malnutrition is a perennial problem.
Attention should be directed to other areas of cowpea processing and
utilization. More research should be conducted into ways of reducing anti-nutritional
factors, improving IVPD and improving cowpea protiens.
Germinated, roasted and autocalved cowpeas possess adequate nutritional
qualities as protein supplements in cereal-based weaning foods.
134
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