Addis Ababa University

College of Natural and Computational Science

Center for Food Science and Nutrition

Effect of processing on proximate composition and

functional properties of Mung bean [Vigna radiata
(L.)Wilczek] commercial varieties in Ethiopia

By: Wondwossen Alemu

February, 2022
Addis Ababa, Ethiopia

i
 Addis Ababa University

College of Natural and Computational Science

Center for Food Science and Nutrition

Effect of processing on proximate composition and

functional properties of mung bean [Vigna radiata
(L.)Wilczek] commercial varieties in Ethiopia

By: Wondwossen Alemu

Advisor: Prof.Kelbessa Urga

A thesis submitted to, Addis Ababa University College of Natural and

Computational Science Center for Food Science and Nutrition in Partial
Fulfillment for the Masters of food Science and Nutrition

February, 2022
Addis Ababa, Ethiopia

ii
Declaration
I the under signed, declare that this is original work and has never been presented in this or other
universities as well as research center previously and all the source of material used for this
master thesis study have been fully acknowledged.

Name: Wondwossen Alemu Gizaw

Signature: ………………
Date: ……………………

Place: Addis Ababa University, Center for food Science and Nutrition, Addis Ababa, Ethiopia
Date of Submission: ………………
The research paper has been submitted for examination with my approval as University advisors.

Advisor
Prof.Kelbessa Urga
Signature: …………………
Date: ……………………....

i
 Addis Ababa University
College of Natural and Computational Science
Center for Food Science and Nutrition

Effect of processing on proximate composition and functional

properties of mung bean [Vigna radiata (L.)Wilczek] commercial
varieties in Ethiopia

A thesis submitted to the School of Graduate Studies of Addis Ababa University in partial
fulfillment of the requirement for the Degree of Master of Science in Food Science and
Nutrition.

By:
Wondwossen Alemu

Advisor: Prof.Kelbessa Urga

Approval by examining broad: Signature Date

Prof.Kelbessa Urga (Advisor) ________________ ___________

Dr.Yohannes Seyoum (External examiner) _______________ ___________

Dr.Dawd Gashu (Internal examiner) _______________ ____________

Dr.Hasset Tamirat (Chairman) _______________ _____________

ii
 Acknowledgements

I would like to thank Ministry of trade and Industry, European Union and Addis Ababa
University for giving and sponsoring me to study my Master degree in food Science and
Nutrition.

I would like to express my deepest gratitude and thanks to my advisor, Kelbesa Urga (prof) for
support and guidance in the manuscript writing as well as for their encouragement, advice, and
concrete support during conducting this thesis work.

I would like to thank the staff member of Ethiopian Commodity Exchange Authority for their
cooperation to give information about Mung bean production area for my study population for
writing this Thesis.

I also extend my profound gratitude to department of Center for Food Science and Nutrition
Department for providing facilities to conduct this research. I am also very thankful to the
technical staffs of the center especially, Mr. Debeb Hailu, for providing all the necessary
laboratory facilities needed for my analysis.

Finally I would like to thank our staff members of Ethiopian Commodity Exchange Authority
Economic Analysis Directorate for giving me their feedback, suggestion, and editing this Thesis.

i
 Table of Contents
Acknowledgements ................................................................................................................................... i
Acronyms .................................................................................................................................................. v
List of Figures .......................................................................................................................................... vii
List of tables ........................................................................................................................................... viii
Abstract ..................................................................................................................................................... ix
CHAPTER ONE ...................................................................................................................................... 1
1. INTRODUCTION ............................................................................................................................... 1
1.1 Background and justification ................................................................................................ 1
1.2 Statement of the problem ...................................................................................................... 2
1.3 Objectives of the proposed study .......................................................................................... 3
1.3.1 General objectives .......................................................................................................... 3
1.3.2 Specific objectives .......................................................................................................... 3
1.4 Significance of the study ....................................................................................................... 3
CHAPTER TWO ..................................................................................................................................... 5
2. LITERATURE REVIEW ................................................................................................................... 5
2.1. Over view of mung bean ...................................................................................................... 5
2.1.1. World production of mung bean.................................................................................... 5
2.1.2 Production of mung bean in Ethiopia ............................................................................. 5
2.2 Mungbean and health ............................................................................................................ 7
2.3 Antinutritional factors in Mung bean .................................................................................... 8
2.3.1Antinutritional effects of phytate ..................................................................................... 8
2.3.2 Antinutritional effects of tannins .................................................................................... 9
2.4 Antioxidants .......................................................................................................................... 9
2.5 Effect of processing methods on mung bean ...................................................................... 10
2.5.1 Boiling .......................................................................................................................... 11
2.5.2 Soaking ......................................................................................................................... 11
2.5.3 Germination .................................................................................................................. 12
2.6 Functional properties ........................................................................................................... 12
2.6.1 Water absorption capacity ........................................................................................... 12
2.6.2 Oil absorption capacity ................................................................................................. 13
2.6.3 Bulk density .................................................................................................................. 13

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 2.6.4 Swelling power and solubility ...................................................................................... 13
CHAPTER THREE ............................................................................................................................... 14
3. MATERIALS AND METHOD ....................................................................................................... 14
3.1 Origin of the mung bean...................................................................................................... 14
3.2 Experimental Design ........................................................................................................... 14
3.3 Sample Collection ............................................................................................................... 14
3.4. Processing methods ............................................................................................................ 15
3.4.1. Raw (control) ............................................................................................................... 15
3.4.2. Dehulling ..................................................................................................................... 16
3.4.3. Soaking ........................................................................................................................ 16
3.4.4. Dry roasting ................................................................................................................. 16
3.4.5. Germination ................................................................................................................. 17
3.4.6. Boiling ......................................................................................................................... 17
3.5. Determination of proximate composition .......................................................................... 18
3.5.1 Moisture ........................................................................................................................ 18
3.5.2 Crude Protein ................................................................................................................ 18
3.5.3 Crude Fat ...................................................................................................................... 20
3.5.4 Total ash ....................................................................................................................... 21
3.5.5 Crude Fiber ................................................................................................................... 21
3.5.6 Utilizable carbohydrate determination ......................................................................... 22
3.5.7 Total energy in kilo calories ......................................................................................... 22
3.6.2 Functional properties of mung bean flour .................................................................... 22
3.7.3 Determination of Antinutritional Factors ..................................................................... 24
3.8.4 Determination of antioxidant activity ........................................................................... 26
3.9. Statistical Analysis ............................................................................................................. 28
CHAPTER FOUR .................................................................................................................................. 29
4. RESULT AND DISCUSSION ......................................................................................................... 29
4.1.Proximate composition of mung bean flour ........................................................................ 29
4.1.1. Moisture .......................................................................................................................... 29
4.1.2. Crude protein ............................................................................................................... 29
4.1.3 Crude Fat ...................................................................................................................... 30
4.1.4 Total ash ....................................................................................................................... 31

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 4.1.5 Crude fiber .................................................................................................................... 31
4.1.6 Utilizable carbohydrate................................................................................................. 32
4.2 Functional Properties of Mung bean flour .......................................................................... 34
4.2.1 Bulk density .................................................................................................................. 34
4.2.2 Solubility index............................................................................................................. 35
4.2.3 Swelling power ............................................................................................................. 35
4.2.4 Water Absorption Capacity .......................................................................................... 36
4.2.5 Oil Absorption Capacity ............................................................................................... 37
4.3 Determination of Antinutritional Factors of Mung bean flour ............................................ 37
4.3.1 Phytate .......................................................................................................................... 37
4.3.2 Tannin ........................................................................................................................... 38
4.4 Antioxidant activities of mung bean flour........................................................................... 39
4.4.1 Yield of extract ............................................................................................................. 39
4.4.2 Determination of radicals scavenging activity of mung bean by DPPH ...................... 40
4.4.3 Determination of Total Phenol ..................................................................................... 42
4.4.4 Determination of total flavonoid .................................................................................. 43
CHAPTER FIVE .................................................................................................................................... 45
5. CONCLUSION AND RECOMMENDATION ............................................................................. 45
5.1. Conclusion.......................................................................................................................... 45
5.2 Recommendations ............................................................................................................... 46
6. REFERENCES ...................................................................................................................... 47
Appendix-1 ............................................................................................................................................. 53
1. ANOVA and Duncan table of proximity analysis ................................................................ 53
Appendix-2 ............................................................................................................................................. 59
2. ANOVA and Duncan table of functional properties ............................................................. 59
Appendix-3 ............................................................................................................................................. 64
3. Standard Curve, ANOVA and Duncan table of Antinutritional factor ................................. 64
Appendix-4 ............................................................................................................................................. 67
4 Graph of percent of inhibition for three type Mung bean of DPPH, Standard curve, ANOVA
and Duncan table of Antioxidant Activity ................................................................................ 67

iv
 Acronyms
ANOVA Analysis of variance
AOAC Association of Official Analytical Chemists
CSA Central Statistical Agency
DPPH 2, 2-Diphenyl–1-picrylhydrazyl
ECX Ethiopian Commodity Exchange
FAO Food and Agriculture Organization
ROS Reactive Oxygen Species
RNS Reactive Nitrogen Species
SOD Superoxide Dismutase
SPSS Statistical Product and Service Solution
WHO World Health Organization

v
 List of Figures
Figure 1: Mung bean (Masho) or green mung bean in Ethiopia…………………………………6
Figure 2: Mung bean area, production, productivity and participant………………… ...........…6
Figure 3: Flow Chart of the undehulled and dehulled mung bean sample flour ……………….15
Figure 4: Milled and sieved mung bean flour………………………………… ............……….16
Figure 5: Germinated mung bean………………………………………………… .............…..17
Figure 6: Boiled mung bean…………………………………………………… ....................…18
Figure 7 Percentage inhibition verse concentration of samples and ascorbic acid at
IC50 by DPPH………………………………………………………………… ..........………..41

vii
 List of tables
Table 1: Mung bean area,production, and small holders by region from 2006 E.C-2010 E.C ....7
Table 2: Proximate composition of raw and processed mung bean Flour………… ..................33
Table 3: Functional properties of mung bean flour………………………………… ............…34
Table 4: Antinutritional contents of raw and processed mung bean flour................... ...............39
Table 5: Percentage of yields of extract by methanol from raw and processed mung bean… ...40
Table 6: Concentration of ascorbic acid and samples at IC50……………………… ............…42
Table 7: Total phenol and flavonoid contents of raw and processed mung bean………… .......44

viii
 Abstract
Mung bean [Vigna radiata (L.)Wilczek] is one of the important legume crops grown from the
tropical to sub-tropical areas around the world. Effect of processing method on the proximate
composition and functional properties of mung bean in Ethiopia was not well addressed because
the crop is not indigenous and well known throughout in the country .In this study effect of
processing methods, dehulling, germinating, dry roasting, soaking and boiling on proximity
composition, antinutritional factor, antioxidant activity, and functional properties of mung bean
[Vigna radiata (L.)Wilczek] collected from Bale, Gonder and North showa part of Ethiopia were
investigated on dry basis. The proximity composition of both the processed and raw mung bean
were determined by AOAC 2016. The total phenolic extract by methanol were determined using
Folin-Ciocalteu methods while total flavonoid content was estimated by using aluminum
trichloride(AlCl3), Free radical scavenging activity was determined by 2,2-diphenylpicryl-1-
picrylhydrazyl activity using ascorbic acid as standards. The moisture content, ash, crude
protein, crude fat, crude fiber ,utilizable carbohydrate and energy of processed and raw mung
bean flour were ranged from 3.73%-9.40%, 2.13-3.47%, 27.65%-30.92%, 1.33%-2.00%, 0.48%-
7.57%, 50.43%-59.97% and 328.41-374.27%Kcal/100g respectively for raw and processed
sample. The antinutritional factors, level of phytate and tannin were, 133.86-190.75µg/100g,
and13.69- 23.71mg/100g respectively for the raw and processed sample. Among the processing
methods used, Germination had significant effect (p<0.05) on reducing antinutritional factors.
Germination significantly (p<0.05) increases the total phenolic and total flavonoid from (14.28
to 125.72) mg Gallic acid equivalent/g and (42.38 to 190.99) mg D-Catechin equivalent/g
respectively. Dry roasting increased the total flavonoid and reduced total phenolic amount from
the raw mung bean flour. The five processing methods had also significant effect on functional
properties of mung bean. Therefore among the five traditional applied processing methods
germination and dry roasting was recommended process. Finally among the three types of
commercial mung bean varieties Bale type mung bean was better than Gonder type and North
showa type mung bean in most of the parameter determined in this study.
Keywords: Mung bean, Proximity Composition, Processing method, Antinutritional factors,
Functional property, Antioxidant Activity, Total phenol, Flavonoid.

ix
 CHAPTER ONE
1. INTRODUCTION

1.1 Background and justification

Mung bean [Vigna radiata (L.)Wilczek] is one of the important legume crops grown from the
tropical to sub-tropical areas around the world (Kumari et al., 2012; Khan et al., 2012). From the
6 million ha of world mung bean production, about 90% is in Asia and among that, India is the
biggest producer where about 2.99 million ha are cultivated (Nair et al, 2013). It is a minor crop
in Australia, China, Iran, Kenya, Korea, Malaysia, the Middle East, Peru, Taiwan and United
States. Apparently, it has been introduced in different regions of Ethiopia like Shewa, Hararge,
Illubabor, Bale, Gamogofa, Tigray, Asosa and Gondar, Farmers in some moisture stress areas
(Gofa, Konso, South Omo zone and Konta) have been producing mung bean to supplement their
protein needs and also effectively use scanty rainfall. It is an important wide spreading,
herbaceous and annual legume pulse crop cultivated mostly by traditional farmers (Ali et al.,
2010). The crop is characterized by fast growth under warm conditions, low water requirement
and excellent soil fertility enhancement via nitrogen fixation (Yagoob, 2014). Fertilization of this
crop occurs through self-pollination without requirement of other pollinators like insects, water
and wind (Rashid et al., 2013). According to Asfaw et al. (2012), the optimum temperature range
for good production of mung bean ranges from 270 C- 300 C. The temperature necessities always
stay above 150 C and the crop requires from 90–120 days for maturity.

The Ethiopian Commodity Exchange (ECX) installed mung beans as the sixth commodity to be
traded on its floor on Wednesday, January 22, 2014 (ecx.gov.et).The decision was made at the
end of July, 2013, after the Board of the Ethiopian Commodity Exchange Authority (ECEA)
gave its approval. The ECEA is the supervisory body of the ECX and has the final say on
whether or not new commodities can enter the trading floor (ecea.gov.et). The ECX commenced
trading operations in April 2008, with coffee, sesame, maize, wheat and pea beans (ecx.gov.et).
There are four quality grades (grade 1,2,3 and 4) for mung beans, different contract based on
origin such as green mung bean Bale type, green mung bean Gonder type, green mung bean
North showa type, etc. and different delivery sites, located in – Addis Abeba; Kombolcha in
South Wollo Zone (376km from Addis Abeba) and Gonder (656 km from Addis Abeba) of the

1
Amhara Region ,Assosa, the capital of the Benishangul Gumuz Region (657 km from Addis
Abeba) (ecx.gov.et).

Although Mung bean is widely known for its fiber, mineral and protein, at present it is
considered not only as a rich source of nutrients but also a source of other bioactive compounds
with many beneficial physiological effects such as antioxidant, antidiabetic, anticholesteromic
and anticancer effect in controlling and preventing various metabolic diseases. Previous studies
done by Bhatty et al. (2000), Mubarak (2005), Huang et al. (2014) and Chandrasiri et al. (2016)
have shown that processing alters the proximate composition and functional properties of food.
Some processing methods can increase and some can decrease the nutritional and functional
properties of food. Therefore a great attention should be paid not only on what is eaten but also
on the way of preparing it. Mung bean is processed into various forms such as sprouted, cooked
and boiled before consumption. However, the effect of processing on nutritional and functional
properties of Mung bean has not been widely studied in Ethiopia. Thus, the objective of this
study is to investigate the effect of dry roasting, dehulling, soaking, germination and boiling on
proximate composition and functional properties of mung bean varieties in Ethiopia.

1.2 Statement of the problem

Effect of processing method on the proximate composition and functional properties of mung
bean in Ethiopia was not well addressed because the crop is not indigenous and well known
throughout in the country. In addition the crop was introduced in Ethiopia ten years ago so that it
is one of the crops that was traded in Ethiopia Commodity Exchange and exported to other
countries (ecx.gov.et). Furthermore, Mung bean was consumed by local population in different
forms.

Some of the previous studies done by Bhatty et al. (2000), Mubarak (2005) Huang et al. (2014)
and Chandrasiri et al. (2016) and available data on the nutrient and anti-nutrient composition of
mung bean do not cover the entire nutrient and anti-nutrient factor where available. This was
because of the possible effects of variety/genetic origin, climate, soil, processing methods,
pesticides and fertilizers on the chemical composition of the mung bean. In addition there was no
information on the nutrient composition and antinutritional factors of mung bean grown in
Ethiopia.

2
Consequently several considerations justify the continued surveillance, knowledge and research
on anti-nutritional factors and toxic substances naturally present in plants used as foods like
mung bean and ways of reducing them to safe level of consumption. Moreover the determination
of minerals and trace elements in foodstuffs was also an important part of nutritional and
toxicological analyses.

1.3 Objectives of the proposed study

1.3.1 General objectives

The general objective of the study was to investigate the effect of processing on proximate
composition antinutritional factor, antioxidant activity and functional properties of mung bean
commercial varieties in Ethiopia.

1.3.2 Specific objectives

The specific objectives of this study were:-

 To determine the proximate composition (moisture content, total ash, crude protein, crude
fiber, and crude fat and utilizable carbohydrate) of raw and processed mung bean.

 To investigate the effect of dry roasting, dehulling, soaking, germination and boiling of
mung bean on proximate composition and functional properties of flours prepared by
these treatments.

 To investigate the effect of processing on antioxidant activity and antinutritional factor of

mung bean.

1.4 Significance of the study

In Ethiopia mung bean grains was used in different form such as green vegetables, „Kollo‟
(soaked and roasted) and „Nifro‟ (boiled seed) and added in soup etc. Furthermore in all forms, it
may be consumed alone or mixed with other cereals. The processing method used to convert
mung bean in to consumable forms including dehulling, soaking, roasting, boiling, germinating,
oven cooking etc. The results of this study can be used to promote the consumption of the
legume seeds of mung bean by enhancing the bioavailability of minerals to prevent micronutrient
malnutrition.

3
 The finding helps to promote in which processing method is best to get the good quality
nutrient that are found in mung bean.

 The report helps to promote the inclusion of mung bean in children formula food in
Ethiopia.

 The finding may be used as source of information for the public, researchers etc.

4
 CHAPTER TWO

2. LITERATURE REVIEW

2.1. Over view of mung bean

2.1.1. World production of mung bean

The center of origin of cultivated mung bean is Asia. As Sangsiri et al. (2007, 2009) reported,
mung bean originated in the India-Burma region of Asia because evidence provided by
archaebotanical findings and literary records showed that mung bean was domesticated in India
where wild mung bean is widely distributed. Specifically, it has been found at Neolithic sites in
southern India where wild mung bean exists. The findings also point to both south-eastern and
western Himalayan foothills as likely places where domestication could have taken place. Altaf
(2009) also suggested that the primary gene diversity center for mung bean was the central Asian
Region with India having the widest diversity of domesticated varieties of the crop. Mung bean
is one of the important legume crops grown from the tropical to sub-tropical areas around the
world (Kumari et al., 2012; Khan et al., 2012). From the 6 million ha of world mung bean
production, about 90% is in Asia and among that, India is the biggest producer where about 2.99
million ha are cultivated (Nair et al, 2013). It is a minor crop in Australia, China, Iran, Kenya,
Korea, Malaysia, the Middle East, Peru, Taiwan and United States.

2.1.2 Production of mung bean in Ethiopia

Mung bean production in Ethiopia was introduced recently so that Ethiopian Statistical Agency
(CSA) included to report in its annual agricultural sample survey since 2006 E.C. From the CSA
annual agricultural sample survey report since 2006E.C-2010 E.C for the five-year report
indicated that area in hectare increased from 10700ha to 41633.3ha and production increased
from 80600 quintal to 514220 quintal. In addition productivity increased from 7.54qu/ha to
12.35qu/ha. Furthermore small holder farmer in 2006 E.C was estimated to 90437 but in 2010
E.C estimated to be 325788(Fig 2).

5

Fig 1: Mung bean (Masho) or green mung bean in Ethiopia

Mung bean area,production,productivity and

small holders
450000
400000
350000
300000
250000
200000
150000
100000
50000
0
2006 E.C 2007 E.C 2008 E.C 2009 E.C 2010 E.C

Area in hectare Production in quintal Productivity Small holders

source CSA anuall agricultural survey

Fig2: Mung bean area,production, productivity and small holders

From fig 2 we can seen that the area,production, productivity and small holders increased
through out the last five year production year.Furthermore, in Ethiopia mung bean has produced
in some of the regions according to Ethiopian Central Statistical Agency annual agricultural
sample survey report since 2006 E.C-2010 E.C was listed by region in Table 2.1.From this we
can seen that most of the prodcution of mung bean is produced in Amahara region its
area,production and small holders was 88.25%,96.74% and 82.03% respectively.In addition in
oromia region their was no report in area,production and small holders in 2006 E.C,2007 E.C

6
and 2008 E.C respectively but there was report in area and small holders in 2009 E.C and 2010
E.C so that their contribution in area was 9.6% and in small holders was 11.19%

Table 1: Mung bean area,production, and small holders by region from 2006 E.C-2010 E.C
Region Mung bean production by year Total %

2006 E.C 2007 E.C 2008 E.C 2009 E.C 2010 E.C

Oromia Area in hectare * * * 5711.12 5813.65 11524.77 9.6%

Production in quintal * * * * * 0 0.00%
Small holders 3158.00 * 9022.00 32766.00 38583 83529.00 11.19%
Amhara Area in hectare 9808.22 11281.69 24038.85 28992.86 31670.70 105792.32 88.25%
Production in quintal 80640.10 119828.48 247422.26 352972.50 403014.67 1203878.21 96.74%
Small holders 52802.00 52473.00 120341.00 131991.00 254765.00 612375.00 82.03%
Tigray Area in hectare * * * * * *
Production in quintal * * * * * *
Small holders 1112.00 * * * * 1112.00 0.15%
SNNP Area in hectare * * 224.40 907.61 1132.01 0.94%
Production in quintal * * 2090.10 4722.85
6812.95 0.55%

Small holders * * 4426.00 8827.00 18536.00 31789.00 4.21%

Benshagul Area in hectare * * * 1255.43 1427.64 2683.07 2.24%
Production in quintal * * * 15155.87 18540.93 33696.80 2.71%
Small holders * * * 7099.00 10612.00 17711.00 2.31%
Area in hectare 119876.74 100%

Total Production in quintal 1244387.96 100%

Small holders 746516.00 100%

Source CSA annual agricultural survey report

2.2 Mungbean and health

Mung bean is an important grain legume in South, East and Southeast Asia, which produce up to
three million metric tons of seed consumed directly as dhal, porridge and bean sprouts, or
processed into high value noodles (Nair et al, 2012). To meet global mung bean demand and to
address widespread malnutrition, it is imperative to improve the current average global

7
productivity as well as to expand its reach into new regions, including Central Asia and Africa
(Nair et al,2013). Mung bean is a substantive source of dietary protein (24% - 28%) and
carbohydrates (59% - 65%) on a dry weight basis, and provides about 3400 kJ energy/kg grain
(USDA, 2010). In comparison to other legumes, such as chickpea (Cicer arietinum), pigeon pea
(Cajanus Cajan), and lentils (Lens culinaris), mung bean starch is easier to digest (Sandhu et al,
2008). Mung bean also induces less flatulence and is well tolerated by children (Pal et al, 2010).
In addition, mung bean is lower in phytic acid (72% of the total Phosphors content) than pea,
soybean (Glycine max), and cereals. Phytic acid is commonly found in cereal and legume crops
and has a negative impact on iron and zinc bioavailability in plant-based human diets. Owing to
its palatable taste and nutritional quality, mung bean has been used as an iron-rich whole food
source for baby food (Imtiaz et al, 2011).
The seeds and sprouts of mung bean contain abundant nutrients with biological activities. The
review by Tang et al, 2014 provides insights into the nutritional value of mung beans.
Constituents that have been isolated in the past few decades, such as flavonoids, phenolic acids,
organic acids, amino acids, carbohydrates, and lipids have been discussed. In addition, dynamic
changes in metabolites during the sprouting process and related biological activities, including
antioxidant, and health promoting effects, are evidence of its use as a medicine.

2.3 Antinutritional factors in Mung bean

An antinutrient is a substance occurring in the diet which acts antagonistically towards one or
multiple nutrients, reducing bioavailability. This is usually done through complex formation
which reduces nutrient absorption (Graham et al., 2000).

2.3.1Antinutritional effects of phytate

Phytic acid (phytate; myo-inositol 1, 2, 3, 4, 5, 6-hexakisphosphate) is the primary source of
inositol and storage phosphorus in plant seeds contributing ~ 70% of total phosphorus. The
abundance of phytic acid in cereal grains is a concern in the foods and animal feeds industries
because the phosphorus in this form is unavailable to monogastric animals due to a lack of
endogenous phytases; enzymes specific for the dephosphorylation of phytic acid. In addition, the
strong chelating characteristic of phytic acid reduces the bioavailability of other essential dietary
nutrients such as minerals (e.g. Ca2+, Zn2+, Mg2+, Mn2+, Fe2+/3+), proteins and amino acids(Walter

8
et al., 2002). High phytic acid content feeds are generally supplemented with inorganic
phosphate; however this causes increased fecal phosphate levels and subsequent eutrophication
of waterways. Alternatively, supplementation with commercial phytases is becoming
increasingly popular and reduces the requirement for inorganic phosphate supplementation as
well as the associated environmental issues. The daily intake of phytate was estimated to be
2000–2600 mg for vegetarian diets as well as diets of inhabitants of rural areas of developing
countries and 150–1400mg for mixed diets.

2.3.2 Antinutritional effects of tannins

Tannins are polyphenols components prevalent in food legumes. Tannins are mainly located in
seed coat of pulses, hence physical removal of seed coat by either dehulling or milling and
separating hulls decreases the tannin content of pulses and improves their nutritional quality.
Dehulling eliminates about 68 to 99% of tannins in seed. Soaking of seeds before cooking is a
common household practice and, used to soften the texture and hasten the process of cooking.

Studies have shown that tannins interact with proteins, enzymes or nonenzymes, and form
tannin-protein complexes, which decrease protein digestibility and protein solubility. This
decrease in protein digestibility may be caused by either the inactivation of digestive enzyme or
the reduction of the susceptibility of the substrate proteins after forming the complex.
Polyphenols are found to interact with proteins and cause either inactivation of enzyme such as
trypsin and chymotrypsin or make protein insoluble. Polyphenols inhibit several enzymes
including α-amylase, lipases, pectin esterase, cellulase and β-galactosidase (Salunkhe et al.,
1985). The total acceptable tannin daily intake for human being is 560mg/100g ml (Shimelis and
Rakshit, 2007).

2.4 Antioxidants

Antioxidants are scavenging molecules that convert reactive oxygen species (ROS) to water (H2O)
to prevent overproduction of reactive oxygen species. There are two types of antioxidants in the
human body: enzymatic antioxidants and non-enzymatic antioxidants (Domej et al, 2014).
Enzymatic antioxidants – are also known as natural or endogenous antioxidants as stated above.
They neutralize excessive ROS and RNS and so prevent them from damaging the cellular structure

9
Enzymatic antioxidants, which help to prevent the excessive accumulation or upsurge of free
radicals are composed of superoxide dismutase (SOD), catalase, glutathione peroxidase and
glutathione reductase, which also causes reduction of hydrogen peroxide to water. As stated
above hydrogen peroxide is not a free radical but it is a potentially toxic oxidant and so must be
eliminated from the body. The hydrogen peroxide is eliminated by the action of catalase to give
water and oxygen from two molecules of hydrogen peroxide. Hydrogen peroxide can also be
eliminated by the action of the enzyme glutathione peroxidase in which hydrogen peroxide reacts
with NADPH + H+ to form NADP and water (Halliwell &Gutteridge, 2002).

Non-enzymatic antioxidants – are also known as synthetic antioxidants or dietary or natural-

occurring antioxidant vitamins. The body‟s complex antioxidant system is influenced by dietary
intake of antioxidant vitamins and minerals such as ascorbic acid or vitamin C, tocopherols and
tocotrienols or vitamin E, selenium, zinc, taurine, hypotaurine, glutathione, beta carotene and
carotene (provitamin A). Vitamin C or ascorbic acid is a chain breaking antioxidant that stops the
propagation of the peroxidative process and also helps to recycle oxidized vitamin E and
glutathione (lobo et al, 2010).

Phytochemicals are compounds which are found in vegetables together with fruits and legumes
(i.e. they are rich in plant-based foods). Though these phytochemicals are found in very small or
minute amounts they act as very potent or effective antioxidants. They advance or promote
health and help in the prevention of coronary heart disease (keep cholesterol from depositing on
the walls of the arteries and thus, lower the progress of arteriosclerosis) and cancer (Podmore et
al, 1998).

2.5 Effect of processing methods on mung bean

The effects of some domestic traditional processes, such as dehulling, soaking, germination,
boiling, autoclaving and microwave cooking, on the nutritional composition and anti-nutritional
factors of mung bean seeds were studied. Germination and cooking processes caused significant
(p < 0:05) decreases in fat, carbohydrate fractions, anti-nutritional factors and total ash contents.
All processes decreased the concentrations of lysine, tryptophan, threonine and sulfur-containing
amino acids. However, all treatments were higher in total aromatic amino acids, leucine,
isoleucine and valine contents than the FAO/WHO reference (El-Adawy, 2002). Dehulling,
soaking and germination processes were less effective than cooking processes in reducing trypsin

10
inhibitor, tannins and hemagglutinin activity contents. Also, germination was more effective in
reducing phytic acid, stachyose and raffinose. Germination resulted in a greater retention of all
minerals compared to other processes. In vitro protein digestibility and protein efficiency ratio
were improved by all processes. The chemical score and limiting amino acids of mung bean
subjected to the various processes varied considerably, depending on the type of process (El-
Beltagy, 1996).

2.5.1 Boiling

Boiling/cooking and roasting are important food processing methods. As a thermal process,
boiling/cooking could enhance the palatability and nutritional value by inactivating endogenous
toxic factors. Roasting is similar to cooking/boiling but involves higher temperature and reduced
time. Boiling is effective method in reducing water soluble antinutrient. Boiling is also found to
decrease some amount of soluble phytate. Thus, providing plants with heat-stable phytases or
addition of exogenous heat-stable phytases are seen as possibilities to improve phytate
dephosphorylation during cooking (Greiner and Konietzny, 1998).

2.5.2 Soaking

Soaking is often used as a pretreatment to facilitate processing of legume grains and cereal seeds.
Soaking may last for a short period, about 15 to 20 minutes, or for a very long period, usually 12
to 16 hours. In household situations cereals and legumes are typically soaked in water at room
temperatures overnight. Because phytate is water soluble, a significant phytate reduction can be
realized by discarding the soak water. In addition, action of endogenous phytases contributes to
phytate reduction. Temperature and pH value have been shown to have a significant effect on
enzymatic phytate hydrolysis during soaking. If the soaking step is carried out at temperatures
between 45 and 65 °C and pH values between pH=5.0 and 6.0, which are close to the optimal
conditions for phytate dephosphorylation by the intrinsic plant phytases, a significant percentage
of phytate (26–100 %) was enzymatically hydrolyzed (Greiner and Konietzny, 2006).

Moreover, soaking removes some of the non-nutritional compounds, which can be partly or
totally solubilized and eliminated with the discarded soaking solution (Frias et al., 2000). Some
previous studies reported in the literature have analyzed the effect of the soaking conditions on
the main quality parameters in the final product (Frias et al., 2000; Sayar et al., 2001). Frias et al
(2000) concluded that the seed coat controls water absorption up to a certain level of moisture.

11
Frias et al. (2000) also concluded that the differences in water absorption might be due to the
differential solubilization of the starch during soaking which is caused mainly by differences in
starch structure, seed size and membrane permeability.

2.5.3 Germination

Germination is a process widely used in legumes and cereals to increase their palatability and
nutritional value, particularly through the breakdown of certain antinutrient, such as phytate and
protease inhibitors. In non-germinated legume grains and cereal seeds, with the exception of rye
and to some extent wheat, triticale and barley, only little intrinsic phytate-degrading activity is
found, but during germination a marked increase in phytate-degrading activity with a
concomitant decline in phytate content was observed (Graham et al., 2000).

Phytate is hydrolyzed during germination in a stepwise manner by phytases or a concerted action

of phytases and phosphatases which do not accept phytate as a substrate to supply the nutritional
needs of the plant without an accumulation of less phosphorylated myo-inositol intermediates.

2.6 Functional properties

Functional properties are very important in determining the level of utilization in ingredient
formulation and new food product development (Fasasi, 2007). Before consideration is given to
mung bean as potential sources of flour and starch to produce foods, it is necessary to
characterize their chemical composition, physical, physicochemical, and functional properties
(Elevina et al., 2010). The chemical composition of flours and starches exhibits differences
especially in amylose and phosphorous content, as a function of the botanical origin. It is
significant because of the influence of amylose and phosphorous content in the functional
properties of flours and starches. It is a general consensus that the influence of both amylose and
phosphorous content affects the gelatinization and pasting behavior of starches and flours. These
two parameters determine the functional properties of flours and starches such as: texture,
consistency, binding, coating, adhesiveness, cohesiveness, thickening, viscosity, and palatability
(Elevina et al., 2010).

2.6.1 Water absorption capacity

The ability to absorb water is a very important property of flours used in food preparation. The
ability of food materials to absorb water is sometimes attributed to the protein content (Mbofung,
2006). Water absorption capacity is an important functional property required in food

12
formulations especially those involving dough handling (Udensi and Okoronkwo, 2006). WAC
plays a major role in the functionality of dough. In particular, WAC has been shown to be related
to dough consistency (Njintang, et al., 2008). It is known that water binding by starches and
flours is a function of several parameters including size, shape, conformational characteristics,
steric factors, hydrophilic hydrophobic balance in the starch molecule, lipids and carbohydrates
associated with the proteins, thermodynamic properties of the system (energy of bonding,
interfacial tension, etc.), physicochemical environment (pH, ionic strength, vapor pressure,
temperature, presence/absence of surfactant etc.), solubility of starch molecules and others
(Shimelis,2006).

2.6.2 Oil absorption capacity

Oil absorption capacity (OAC) is defined as the difference in the flour weight before and after its
oil absorption (Giami et al., 1994). It is great importance, since fat acts as flavor retainer and also
increases soft texture to mouth feel of foods, especially bread and other baked foods (Ubbor and
Akobundu, 2009). They are also important because of their storage stability and particularly in
the rancidity development.

2.6.3 Bulk density

Bulk density gives an indication of the relative volume of packaging material required.
Generally, higher bulk density is desirable for the greater ease of dispersibility and reduction of
paste thickness which is an important factor in convalescent child feeding (Udensi and
Okoronkwo, 2006).

2.6.4 Swelling power and solubility

Swelling power provides evidence of non-covalent bonding between starch molecules. Factors
like amylose-amylopectin ratio, chain length and molecular weight distribution, degree/length of
branching and confirmation determine the degree of swelling and solubility. Solubility of flours
depends on a number of factors such as sucrose, interassociative forces, swelling power,
presence of other factors, etc. (Subramony, 2002).

13
 CHAPTER THREE
3. MATERIALS AND METHOD

3.1 Origin of the mung bean

The origin of the mung bean used in the study were North Showa, Gonder, and Bale in this area
more than 96 % of the mung bean production produced and exported in Ethiopia.

3.2 Experimental Design

Completely randomized design were used to study the effect of processing (dehulling, soaking,
dry roasting, boiling and germination ) on proximate composition, anti-nutrients (phytate,
tannin),antioxidant activity(DPPH, flavonoid and phenol) and functional properties (BD, WAC
and OAC) of mung bean flours were studied.

3.3 Sample Collection

Six kilogram of each North Showa type, Bale type and Gonder type were collected from
Ethiopian Commodity Exchange of Gonder and Saris warehouses. The seeds were cleaned
manually by removing any foreign material, damaged and broken seeds, shriveled and insect
attacked seeds. The seeds were processed by direct grinding of raw (used as control), dehulling,
soaking, germination, boiling and dry roasting. The processed samples except the roasted one
were dried in an oven at 50°C for 24 h. All the samples including the control were ground by a
laboratory mill to pass through a 0.05μm sieve and was kept in moisture proof plastic bag placed
in air tight tin container at 40 0C. The seed flours of both the control and processed samples were
evaluated for nutritional composition, anti-nutritional and functional properties.

14
 Whole mung
bean

Dry Cleaning

Milling

Dry roasting Sieving Soaking Germinating

Soaking
Milling Raw flour Boiling Oven drying Oven drying
Dehulled Oven drying Milling

Oven drying Sieving Milling Milling

Sieving
Milling Sieving Sieving
Roasted flour
Soaked flour
Sieving
Boiled flour
Dehulled flour Germinated flour

Fig 3: Flow Chart of the undehulled and dehulled mung bean sample flour

3.4. Processing methods

3.4.1. Raw (control)

Cleaned seed of 500 g of each of the four mung bean type samples were directly ground by a
mill (Teklehaimanot et al., 1993).

15
Fig 4: Milled and sieved mung bean flour

3.4.2. Dehulling
Hulls were removed manually after soaking 600 g cleaned seeds of the three mung bean types for
6 h in distilled water at room temperature. Seeds were completely covered by water using seed to
water ratio of 1:3 (w/v) (Nestares et al., 2003). The dehulled seeds were then dried and milled to
flour.

3.4.3. Soaking
Cleaned 500 g each of the three mung bean types were soaked for 12 h in distilled water at room
temperature. Seeds were completely covered by water using seed-to-water ratio of 1:3 (w/v)
(Nestares et al., 2003)

3.4.4. Dry roasting

Cleaned 500 g seeds each of the three mung bean types were roasted by hot air oven for 30
minutes at 150°C.The cooled samples were then milled in to flour, (Suvendu and prakash, 1997).

16
3.4.5. Germination
Cleaned 500 g each of the three mung bean types were washed and cleaned with tap water.
Germination was done according to the method used by Shimelis and Rakshit, (2005). It was
performed at room temperature in a dark room. Washed mung bean type were soaked for 12 h in
distilled water using seed to water as 1:5(w/v). Soaked seeds were spread on moist filter paper on
large plastic screen. The seeds were then cover with filter paper to reduce evaporation. The
screen was placed in perforated plastic container and keep in the dark at room temperature for 72
h to germinate. The seeds were splashed every 24 h with running sodium hypochlorite at
concentration of 0.01% (w/v) for 10 min to avoid mold contamination. At the end of germination
period, non-germinated seeds were discarded and the germinated ones were dried at 50oC for 24
h. The dry germinated seeds were milled to flour.

Fig 5: Germinated mung bean

3.4.6. Boiling
Cleaned 500 g seeds of the three mung bean types were washed under tap water, were rinsed
with distilled water, and placed in 2 L of distilled boiling water at 96°C and was cooked for 60
min. (until soft) (Teklehaimanot et al., 1993). The boiled samples were dried and milled.

17
Fig 6: Boiled mung bean

3.5. Determination of proximate composition

Moisture, crude protein, crude fat, ash, crude carbohydrate, crude fiber and total energy content
of the three processed mung bean types were determined by using AOAC official 2016.

3.5.1 Moisture
Moisture was determined according to AOAC official 2016. A clean dried and covered flat
aluminum dishes were weighed and about 5gm of the sample was transferred to the dish. The
dish then was placed in an oven set at 1050C for 3hrs and then cooled in desiccators and re-
weighed. Then, the moisture content was estimated by the formula:-
Moisture content in percent (%) = (W2-W3)*100
(W2-W1)
Where W1= weight of crucible
W2= weight of crucible and sample
W3=weight of crucible and sample after dried

3.5.2 Crude Protein

3.5.2.1 Sample Preparation

The samples were weighed of 0.5gm of processed and whole mung bean of the three types in a
Tecator tube and we placed it in the Tecator rack. Then we added 6ml of concentrated Sulfuric
acid from Bird or oxford pipette then after immediately mixed the sample and acid carefully.
Then we were added 3.5ml of Hydrogen peroxide step by step then watched for violent reactions

18
as soon as the most violent reaction had ceased we shacked the tube a few time by hand and we
put it back in the rack. Finally we added 3 gram of mixture of copper sulphate and potassium
sulphate catalyst and waited for 5-15 minutes before digestion.

3.5.2.2 Digestion
We placed sample (accurately weighed) in Kjeldahl flask and with temperature of the digester at
370 0C lower the tubes (in the rack) in to the digester and we continued the digestion until clear
solution was obtained about 4 hours .Then transfer the tubes in the rack in to fume hood cooled
and finally we added 50 ml of water and shacked to avoid precipitation of sulphate in the
solution.

3.5.2.3 Distillation
We added 25 ml of the 35% of sodium hydroxide solution in to the digested and diluted solution
then we placed 250 ml conical flask containing 25 ml of the boric acid, 25 ml of distilled water
and indicator solution under the condenser of the distiller with its tip immersed in to the solution.
Finally distillation would continue until total volume became between 200ml and 250ml and
rinsed the tip with a few ml of water before the receiver is removed.

3.5.2.4 Titration
First we prepared 0.1N HCl in 1000 ml of volumetric flask by using dilution formula as
followed:

19
 M=%concentration *specific density *10 37%*1.18*10 =12N
Molecular weight of HCl 36.5
Normality = molarity in HCl
By using dilution formula C1V1=C2V2
V1=C2V2 = 0.1N*1000ml = 8.3ml
C1 12N

We had taken 8.3 ml of concentrated HCl and diluted with distilled water in 1000ml of
volumetric flask .Then titrate with 0.1 N HCl until we seen a reddish color.

Mole of HCl =Moles of NH3=Moles of N in the sample

Then the crude protein content was determined using the following equation
Nitrogen (%) = VHCl in L *0.1 N HCl*corrected acid volume *14*100
W (weight of sample )

The conversion factor is 6.25, which is obtained from food composition table of EPHI.

Crude protein content (%) = total nitrogen (%) × 6.25

3.5.3 Crude Fat

First we washed extraction cylinder with hot water to remove any impurity and put it into an
oven for about 1 hour at a temperature of 105 oC and we took out it and put them into a
desiccator and weighed (W1) and we put it again in the desiccator. Then we covered the bottom
of an extraction thimble with a layer of fat free cotton or defatted cotton and weighed about 2 gm
of the sample in the thimble (W) accurately and covered with a layer of fat free cotton or defatted
cotton after that we were put the thimble in the extraction chamber. An extraction cylinder from
the desiccator has been taken out and put it on the bracket, to check the number and 50 ml of
petroleum ether has been added in to the extraction cylinder and move in to the heating plank to
late the extraction go on for 4 hours. Then the extraction cylinder was disconnected and put it in
the drying oven at 70 0C for 30 minute and we put it in the desiccator to cool for half an hour.

20
Finally the sample was weighed the extraction cylinder immediately after it was taken out of the
desiccator (W2).

Then the crude fat, percent content was determined using the following equation
Crude fat, percent by weight (%) = (W2-W1)*100
W
Where : W1= weight of the extraction flask (g)
W2= weight of the extraction flask plus the dried crude fat(g)
W= weight of sample (g)

3.5.4 Total ash

We had cleaned a Porcelain crucible and dried it in a muffle furnace for 30 min at 550 0C and
cooled the crucible in a desiccators (with granular silica gel) for about 30 minutes or more at
room temperature and weighed it (W1) then we weighed about 2.5 g of fresh sample to an
accuracy of 4 decimal places in the crucible (W2) chare the sample on a hot plate under a fume –
hood and slowly increased the temperature until smoking ceases. Next we were ash the sample
in the muffle furnace at 550 0C for 5 hours and the ash should be clean and white in appearance
and finally cooled to room temperature and reweighed (W3) each crucible with ash. Then the ash
content was determined using the following equation
Total ash %=W3-W1 *100
W2-W1
Where :W1=weight of the crucible
W2=weight of crucible and sample
W3= weight of crucible and sample after ashing
( W2-W1) is sample mass in g on dry base and (W3-W1) Mass of ash in g

3.5.5 Crude Fiber

Crude fiber analysis was conducted using the method of AOAC official 2016.About 1.6g
weighed sample was transferred into a 600 ml beaker and about 200 ml 1.25% sulfuric acid was
added and boiled for 30 minutes. Recording took place by placing a watch glass over the mouth
of the beaker. After 30 minutes heating by gently keeping the level constant with distilled water,
20 ml 28% KOH was added and boiled gently again for another 30 minutes. Subsequently,

21
washing was conducted with 1% sulfuric acid and NaOH solution. After, filtering it was then
dried in an electric oven for 2hrs. Furthermore, it was cooled at room temperature for 30 minutes
in a desiccators and weighed, then transferred the crucibles to muffle furnace for 30 minute
ashing at 5500C. Finally, it was cooled again in desiccators and reweighed. The crude fiber
content was determined by using the formula:-

%Crude Fiber= W2-W3 *100

W1
Where W1=Crucible weight after drying
W2=crucible weight after ash,
W3=dry weight

3.5.6 Utilizable carbohydrate determination

The total utilizable carbohydrate was calculated by difference with the exclusion of crude fiber.
Total carbohydrate (%) = 100 - (crude fat + crude fiber + crude protein + ash + moisture)

3.5.7 Total energy in kilo calories

The gross energy (GE) content in each sample was determined mathematically using the
following formulae:
Gross energy (Kcal) = (9 x crude fat) + (4 x crude protein) + (4 x utilizable carbohydrate).

3.6.2 Functional properties of mung bean flour

3.6.2.1. Bulk density

Bulk density was determined by the method of Narayana and Narasinga-Rao (1984). An empty
calibrated centrifuge tube was weighed. The tube was filled with a sample to 5 ml by constant
tapping until there was no further change in volume. The weight of the tube and its contents was
taken and recorded. The weight of the sample was determined by difference. Bulk density was
calculated as weight per unit volume of the sample.

22
Bulk density, g/cm3 = W2-W1
Vol.of sample after tapping

W1= weight of tube in g

W2= weight of tube with sample in g

3.6.2.2. Solubility and swelling power

The method used by Tester and Morrison (1990) and Anderson et al. (1969) was used to
determine the solubility and swelling power. About 1g ground sample (< 60 mesh) was
suspended in 10 mL of water and was incubated in a thermostatically controlled water bath at
95oC in a tared screw cap tube of 15 mL The suspension was stirred intermittently over 30 min
period to keep the starch granules suspended. The tubes were rapidly be cooled to 20oC. The cool
paste was centrifuged, at 2200 x g for 15 min to separate jell and supernatant. Then, the aqueous
supernatant was removed and poured in to dish for subsequent analysis of solubility pattern.
After this, the weight of the swollen sediment was determined. Supernatant liquid (dissolved
starch) was poured into a tarred evaporating dish and put in air oven at 100oC for 4 h. Water
solubility index was determined from the amount of dried solids obtained the following equation.

Solubility (%) = W1*100

Ws (1-Mc)
Swelling power was calculated by the following equation

Swelling power (%) = W2*100

Wdm (100-solublity)
Dry matter weight = ws (1-Mc)
Where:
W1= Weight of dissolved solids in supernatant, g
W2= Weight of centrifuged swollen granules, g
Ws= Weight of sample, g
Mc= Moisture content of sample, dry basis (decimal), g
Wdm= Weight of dry matter, g

23
3.6.2.3. Water absorption capacity (WAC)
Water absorption capacity was determined using the method of Beuchat (1977). One gram of the
sample was mixed with 10 ml distilled water for 30 s. The sample was allowed to stand at room
temperature for 30 min and then centrifuged at 5000 x g for 30 min, and the freed water was
taken into a 10 ml graduated cylinder and the volume was recorded. Water absorption capacity
was estimated as the amount of water retained by 100 g materials on dry basis. Density of water
was assumed to be 1 g/ml. The mean of triplicate determinations was reported on a dry weight
basis.

WAC = Weight of water bound

Weight of sample (dry basis)

3.6.2.4. Oil absorption capacity (OAC)

An oil absorption capacity was determined using the method of Beuchat (1977). About one gram
of the sample was mixed with 10 ml oil for 30 sec in a mixer. The samples was allowed to stand
at room temperature for 30min, centrifuged at 5000 x G for 30 min. The freed oil was decanted
into a 10 ml graduated cylinder and the volume was recorded. Oil absorption capacity was
expressed as the amount of oil bound by 100 grams dry matter. Density of oil was determined to
be 0.893 g/ml. The mean of triplicate determinations results was reported on a dry weight basis.
OAC= Weight of oil bound
Weight of sample (dry basis)

3.7.3 Determination of Antinutritional Factors

3.7.3.1 Determination of Phytate content

Phytate was determined by the method of Latta and Eskin (1980) and later modified by Vantraub
and Lapteva (1988). About 0.2g of fresh samples were extracted with 10ml 0.2 % HCl in a
mechanical shaker (Eberbach) for 1hour at an ambient temperature and centrifuged at 3000rpm
for 30 minute. The clear supernatant was used for Phytate estimation. A 2ml of Wade reagent
(containing 0.03% solution of FeCl3.6H2O and 0.3% of sulfosalicilic acid in water) was added to
3ml of the sample solution (supernatant) and the mixture was mixed on a Vortex for 5 seconds.

24
The absorbance of the sample solutions was measured at 500 nm using UVVIS
spectrophotometer.
A series of standard solution was prepared containing 4, 8, 16, 24 and 32 ppm of phytic acid
(analytical grade sodium phytate) in 0.2N HCl. A 3ml of standard was added into 15ml of
centrifuge tubes with 3ml of water which were used as a blank. A 2ml of the Wade reagent was
added to each test tube and the solution was mixed on a Vortex mixer for 5 seconds. The
mixtures were centrifuged for 10 minutes and then absorbance of the solutions (both the sample
and standard) were measured at 500nm by using deionized water as a blank. A standard curve
was made from absorbance versus concentration and the slope and intercept were used for
calculation.
Phytic acid in g/g= [(As-Ab)-Intercept]*10
Slope *w*3

Where;
As= sample absorbance
Ab= blank absorbance
W=Weight of sample

3.7.3.1 Determination of Tannin content

Tannin content was determined by the method of Burns (1971) as modified by Maxson and
Rooney (1972). About 2.0 gram of mung bean flour was weighed in a screw cap test tube. The
mung bean flour was extracted with 10ml of 1% HCl in methanol for 24 hours at room
temperature with mechanical shaking. After 24 hours shaking, the solution was centrifuged at
1000rpm for 5 minutes. A 1ml of supernatant were taken and mixed with 5 ml of vanillin-HCl
reagent (prepared by combining equal volume of 8% concentrated HCl in methanol and 4%
Vanillin in methanol). D-catechin was used as standard for condensed tannin determination. A
40mg of D-catechin were weighed and dissolved in 1000 ml of 1% HCl in methanol, which were
used as stock solution. A 0, 0.2, 0.4, 0.6, 0.8 and 1 ml of stock solution were taken in test tube
and the volume of each test tube was adjusted to 1ml with 1% HCl in methanol. A 5ml of
vanillin-HCl reagent were added into each test tube. After 20 minutes, the absorbance of sample
solutions and the standard solution were measured at 500nm by using water to zero the

25
spectrophotometer, and the calibration curve were constructed from the series of standard
solution using SPSS-22. A standard curve was made from absorbance versus concentration and
the slope and intercept were used for calculation.

Calculation:

Concentration of tannin was read in mg of D-catechin per 100g of sample

Tannin in mg/100g = ((absorbance-intercept)*10)/ (slope x density x weight of sample)

3.8.4 Determination of antioxidant activity

3.8.4.1 Sample extraction

Samples were extracted based on the procedures as outline by Latta and Eskin.The Mung bean
flour samples were homogenized and weight in 5g was then extracted by 25ml of methanol at
250C at 150rpm for 24hour using an incubator shaker (ZHWY-103) and then filtered through
what man No.1 filter paper. The residue was then extracted with additional 25ml of methanol as
described above. The combined methanolic extracts were evaporated at 400c dryness using rotary
evaporator and redissolved in methanol at concentration of 50mg/ml and stored at 40 0c for
further use.

3.8.4.2 Determination of DPPH radical scavenging capacity

DPPH scavenging activity of the tuber methanolic extract was measured according to the method
of Latta and Eskin .IC50 values of the extracts and concentration of the extracts necessary to
decrease the initial concentration of DPPH by 50% were calculated. The hydrogen donation
ability of the corresponding extracts and some pure compound was measured from the bleaching
purpled colored methanol solution of DPPH. The effect of methanolic extract on DPPH radical
was estimated according to Kirby A.J and Schmidt R.J 4ml 0.004% solution of DPPH radical
solution in methanol was mixed with 1ml of various concentrations (2-12mg/ml) of extracts in
methanol with vortex mixer. Incubate the sample for 30min in dark at room temperature
Ascorbic acid standards prepared by dissolving 0.3mg into 1ml of methanol (or 3mg in 10ml
methanol).Scavenging capacity was read Spectrophotometrically by monitoring the decrease in

26
absorbance at 517 nm by (UV-7804C, UV-Vis spectrophotometer. Inhibition of free radical
DPPH in percent (I %) was calculated in following way:
I%= Ac-As * 100
Ac
Where:
Ac=is the absorbance of control reaction without test sample
As =is the absorbance of the test sample (containing all reagents except the test sample) Ascorbic
acid
%I= Percent of inhibition
The concentration of scavenging activities at IC50 was calculated using the %I from the absorbance of
control and absorbance sample solution.

3.8.4.3 Total polyphenol content

The total phenol content was determined by the method described by Siddhuraju, P. and Becker,
K. Aliquots (100 μl) of each extracts were taken in test tubes and made up to the volume of 1 ml
with distilled water. Then 0.5ml of Folin-Ciocalteu phenol reagent (1:1 with water) and 2.5ml of
sodium carbonate solution (20%) were added sequentially in each tube. Rapidly after vortexing
the reaction mixture, the test tubes were placed in dark for 40 minutes and the absorbance was
recorded at 765nm against reagent blank using UV-Vis (UV-7804C ultraviolet visible
spectrometer. Gallic acid was used to construct the standard curve, and the result was expressed
mean± standard deviation expressed as milligram of Gallic acid (GAE) equivalent of extract. All
determinations were carried out in triplicate. 0.05g of Gallic acid in 1ml methanol and then dilute
to 10ml with deionized water (5g/L) final stock. Dissolve 0.1, 0.2, 0.5and 1ml with water to
create standards with 20, 40, 60, 80,100,120 and 140mg/L (ppm) concentration respectively.

The total phenolic compound in the extract in Gallic acid equivalent (GAE) was calculated using
the formulae:

Total phenolic content(C) = GAEc* V

Where, GAEc=concentration of Gallic acid equivalent (milligram/ml) from curve

27
V= Total volume of the extract (ml)

W= sample weight (gm)

3.8.4.4 Total flavonoid

Flavonoid contents were determined according to the method of Zhishen et al. An aliquot (150
μl) of each extract or standard solution was mixed with 1.25 ml of deionized water and 75 μl of
5% NaNO2 solution. After 6 min, 150μl of 10% AlCl3.H2O solution was added. After 5 min, 0.5
ml of 1 M NaOH solution was added and then the total volume was made up to 2.5 ml with
double distilled water. Prepare 10-1000μL quercetin standard in methanol from 1mg/ ml stock by
dissolving (10mg/10ml). Following thorough mixing of the solution, the absorbance against
blank was determined at 510 nm using (UV-7804C, ultraviolet visible spectrometer). The results
were expressed in mg Catechin equivalent (CE).

Flavonoid Content = DE*V

W

Where DE= D-Catechin equivalent (mg/ml)

V= total volume of the sample (ml)

W= sample weight (mg)

3.9. Statistical Analysis

The data which were obtain in this experiment subjected to two ways analysis of variance
(ANOVA) using SPSS version 22 software because in this study we were considered effect of
processing and origin of mung bean as a factor. The mean separation values were determined
using Least significance (LSD) and Duncan multiple range test (DMRT) and significant
differences were defined at p<0.05. The results were presented as mean ± Standard deviation of
three separate determinations.

28
 CHAPTER FOUR

4. RESULT AND DISCUSSION

4.1.Proximate composition of mung bean flour

The proximate composition of raw and processed mung bean sample is presented in Table 2.

4.1.1. Moisture
The moisture contents of raw and processed mung bean flour samples collected from Bale,
Gonder and North showa ranged from 3.73% to 9.40% (Table 2). The maximum moisture
content observed for boiled Gonder type mung bean flour and minimum moisture content
observed for dry roasted Gonder type mung bean flour. The moisture contents for the Bale Type,
Gonder type and North showa type mung bean of the four processing methods, dry roasting,
dehulling and germination, and soaking respectively had significantly decreased moisture
content of mung bean sample as compared to the raw sample(p<0.05). In addition there was no
significant difference of the boiled Bale type, Gonder type and North showa type mung bean as
compared to the raw mung bean(p>0.05). Consequently there were no significant difference
among the moisture content of the Bale type, Gonder type and North showa type mung bean
flour(p>0.05). The interaction of processing methods and origin of mung bean had significant
effect on the moisture content of mung bean flour sample (p<0.05). The relative decrease of
moisture content by processing methods except boiling may be attributed due to a variation in
the treatment during the drying processes of the samples. All the treatments except raw sample
were subjected to drying operation in order to prepare flour for analysis at low moisture content,
the flours possessed low water activity hindering any microbial growth. Moreover, dried flours
are devoid of moisture required for the spore growth and physiological activity. However; other
researchers had earlier reported that raw mung bean had 8.25-10% moisture content (Bhatty et
al., 2000).

4.1.2. Crude protein

The protein content of raw and processed mung bean flour samples ranged from 27.65% to
30.92% (Table 2). The dehulled North showa type samples had highest protein value (30.92 %)

29
and the minimum corresponds to raw Gonder type samples of mung bean (27.65%). The
determinations of crude protein for the Bale Type, Gonder type and North showa type mung
bean of the five processing methods, dehulling, dry roasting and germination, boiling and
soaking respectively had a significantly increased the crude protein content of mung bean sample
as compared to the raw sample(p<0.05). In addition, there was significant difference among the
crude protein content of Bale type, Gonder type and North showa type mung bean flour samples
respectively (p<0.05) but there was no significant difference between the crude protein content
of Gonder type and North showa type mung bean (p>0.05).Consequently the interaction of
processing methods and origin of mung bean had significant effect on the crude protein content
of mung bean flour sample (p<0.05). The results of this study reveal that, dehulling, germination,
dry roasting, boiling and soaking respectively increases crude protein content in mung bean it is
probably due to increasing non-protein nitrogen content during processing by releasing the
bound nitrogenous compounds (e.g.-tannin-protein complexes) (Chandrasiri et al, 2016). The
results obtained were in agreement with those obtained with processed kidney beans (Alonso et
al., 2000) and mung bean (Mubarak, 2005).

4.1.3 Crude Fat

The crude fat content of raw and processed mung bean flour samples were in the range from
1.33% to 2.00% (Table 2). The dry roasted Gonder type samples mung bean flour had highest
crude fat value (2.00 %) and the minimum corresponds to dry roasted Bale type samples of mung
bean flour (1.33%). Furthermore, dry roasting had a significantly increased the fat content of
mung bean flour samples (P < 0.05). In addition soaking, dehulling and boiling had a
significantly reduced the crude fat content of mung bean flour samples (P < 0.05).Moreover,
there was no significant difference between the crude fat content of germinated and raw mung
bean flour samples (p>0.05). Consequently there had a significantly difference between fat
content of Bale, Gonder and North showa type mung bean flours sample(P < 0.05). Finally there
was no significantly difference between the fat content of Gonder and North showa type mung
bean flours (P > 0.05). The interaction of processing methods and origin of mung bean had
significant effect on the crude fat content of mung bean flour sample (p<0.05). The result of this
study shows that a slight decrease in crude fat content of soaked, dehulled and boiled mung bean
flour samples as compared to the raw mung bean flour sample this might be due to their

30
diffusion in water. The fat content of the raw sample (1.5%) was close from the result (1.85%-
2.83%) reported by Bhatty et al. (2000) and Mubarak (2005).

4.1.4 Total ash

The total ash content of raw and processed mung bean flour samples were ranged from 2.13% to
3.47% (Table 2). The maximum ash content observed for dry roasted Gonder type and North
showa type mung bean flour and minimum ash content observed for boiled Bale type mung bean
flour. Moreover, boiling and dehulling had significantly decreased the total ash content of mung
bean flour sample as compared to the raw mung bean flour sample (P < 0.05). Furthermore, there
were no significant difference among the total ash content of germinated, raw, dry roasted and
soaked mung bean flour samples(p>0.05). Consequently there were no significant differences
among the total ash content of the Bale type, Gonder type and North showa type mung bean flour
samples (p>0.05). The interaction of processing methods and origin of mung bean had
significant effect on the total ash content of mung bean flour sample (p<0.05). The result of this
study disclose that there was decreasing in the total ash content of boiled and dehulled mung
bean flour samples as compared to the raw mung bean flour sample. The leaching of minerals
from the seeds during boiling and dehulling could be the reason. It was observed that legumes
contained a large portion of water soluble ash which has the tendency to leach out during hydro
processing of seeds. The raw sample had ash content (3.2%-3.33%) close to the value reported
by Mubarak (2005) for undehulled mung bean seeds flour (3.76%).

4.1.5 Crude fiber

The crude fiber content of raw and processed mung bean flour samples were ranged from 0.48%
to 7.57% (Table 2). The maximum crude fiber content observed for raw Bale type mung bean
flour sample and minimum crude fiber observed for boiled Gonder type mung bean flour sample.
Furthermore, soaking and dehulling, boiling, germination and dry roasting had significantly
decreased the crude fiber content of mung bean flour samples as compared to the raw mung bean
flour sample (P < 0.05). Consequently there were significant differences (p<0.05) among the
crude fiber content of the North showa type, Bale type and Gonder type mung bean flour
samples (p<0.05). Finally there was no significantly difference between the crude fiber content
of Bale and Gonder type mung bean flour sample (P > 0.05). The interaction of processing
methods and origin of mung bean had significant effect on the crude fiber content of mung bean

31
flour sample (p<0.05). The result of this study reveal that all the five processing methods
(soaking, dehulling, boiling, germination and dry roasting) decreased the crude fiber content of
mung bean flour samples as compared to the raw mung bean flour sample this might be due to
solubilization of cellulose and hemi cellulosic material by the processing methods. The crude
fiber content of the raw sample (5.47%-7.57%) was slightly higher than that reported by
Mubarak (2005)-4.63%.

4.1.6 Utilizable carbohydrate

The utilizable carbohydrate content of raw and processed mung bean flour samples were ranged
from 50.43% to 59.97% (Table 2). The maximum utilizable carbohydrate content observed for
dry roasted Bale type mung bean flour sample and minimum utilizable carbohydrate content
observed for raw Bale type mung bean flour sample. In addition, dry roasting, dehulling,
germination and soaking, and boiling had significantly increased the utilizable carbohydrate
content of mung bean flour samples as compared to the raw mung bean flour sample (P < 0.05).
Consequently there were significant differences among the utilizable carbohydrate content of the
North showa type, Bale type and Gonder type mung bean flour(p<0.05). Finally there was no
significantly difference between the utilizable carbohydrate content of Bale and Gonder type
mung bean flour sample (P > 0.05). The interaction of processing methods and origin of mung
bean had significant effect on the utilizable carbohydrate content of mung bean flour sample
(p<0.05). The result of this study reveal that all the five processing methods (soaking, dehulling,
boiling, germination and dry roasting) increased the utilizable carbohydrate content of mung
bean flour samples as compared to the raw mung bean flour sample this might be due to the retro
gradation of starch after gelatinization by the processing methods. This result was in agreement
with the finding of Chandrasiri et al., (2016) which showed that sprouting and boiling increased
in the utilizable carbohydrate content of mung bean flour sample.

32
Table 2: Proximate composition of raw and processed mung bean flour

Total
Method of %Crude %Crude %Crude energy(Kcal/10
processing Origin of mung bean %Moisture %Ash protein %crude Fat Fiber Carbohydrate 0gm)
d cd a*
Bale type mung bean 9.00±0.00 3.20±0.00 28.30±0.20 1.50±0.00b* 7.57±0.30d 50.43±0.50a 328.41±1.20a
Raw Gonder type mung bean
7.47±0.23d 3.33±0.23cd 27.65±0.31a 1.50±0.00b 5.47±0.35d 54.58±0.85a 342.42±2.88a
North showa type mung bean
8.53±0.50d 3.33±0.23cd 28.18±0.31a 1.50±0.00b 6.92±0.05d* 51.54±0.46a* 332.37±2.66a*
Bale type mung bean 5.07±0.23b 3.07±0.23b 30.57±0.20d* 1.50±0.00a* 1.28±0.11ab 58.52±0.20cd 369.86±1.20d
Dehulled Gonder type mung bean
5.33±0.46b 3.07±0.23b 30.09±0.92d 1.00±0.00a 2.06±0.32ab 58.45±1.30cd 367.65±2.04d
North showa type mung bean
5.47±0.50b 3.07±0.23b 30.92±0.40d 1.50±0.00a 1.03±0.16ab* 58.02±0.52cd* 369.25±3.34d*
a d c* c* c d
Bale type mung bean 4.20±0.20 3.20±0.00 30.22±0.71 1.33±0.28 1.07±0.06 59.97±0.80 374.27±0.38d
Gonder type mung bean
Dry roasted 3.73±0.12a 3.47±0.23d 30.15±0.51c 2.00±0.00c 2.29±0.32c 58.36±0.52d 367.54±0.81d
North showa type mung bean
4.47±0.42a 3.47±0.23d 29.69±0.11c 1.50±0.00c 2.74±0.46c* 58.14±0.76d* 364.79±3.46d*

Bale type mung bean 5.07±0.12b 3.33±0.23d 30.28±0.00cd* 1.50±0.00b* 1.23±0.08b 58.59±0.39c 368.97±1.57c
Gonder type mung bean
Germinated 5.53±0.23b 3.47±0.23d 30.19±0.08cd 1.50±0.00b 1.29±0.06b 58.01±0.30c 366.33±1.82c
North showa type mung bean
5.40±0.35b 3.60±0.00d 30.69±0.10cd 1.50±0.00b 1.99±0.24b* 57.81±0.86c* 363.54±1.23c*
Bale type mung bean 7.47±0.23c 3.33±0.23bc 29.52±0.20b* 1.00±0.00a* 0.63±0.10a 58.05±0.10c 363.77±0.39b
Soaked Gonder type mung bean
6.93±0.23c 3.07±0.23bc 29.29±0.10b 1.50±0.00a 1.45±0.17a 57.76±0.15c 361.70±0.69b
North showa type mung bean
6.73±0.23c 3.07±0.23bc 29.40±0.00b 1.50±0.00a 1.74±0.25a* 57.56±0.25c* 361.33±0.99b*
d a c* a* ab b
Bale type mung bean 7.67±0.12 2.13±0.23 30.63±0.31 1.00±0.00 1.19±0.10 57.38±0.30 365.54±1.63b
Boiled Gonder type mung bean
9.40±0.20d 2.27±0.23a 29.98±1.01c 1.50±0.00a 0.48±0.02ab 56.37±1.29b 358.91±1.16b
North showa type mung bean
7.13±0.31d 2.27±0.23a 29.51±0.09c 1.50±0.00a 2.73±0.02ab* 56.87±0.83b* 358.99±2.09b*
All values are the means of triplicates ± standard deviation n=3
Means with the same superscript letters within a column are not significantly different at (P>0.05)
a, b, c, d are superscript to show significance difference between means a<b<c<d
Means with superscript * indicated there is significant different between origin of mung bean (p<0.05)

33
4.2 Functional Properties of Mung bean flour
Table 3. Functional properties of mung bean flour

Water Oil
Method of bulk density Absorption Absorption Swelling Solubility
processing Origin of mung bean Capacity Capacity power Index
Bale type mung bean 1.08±0.00e 1.95±0.05b 1.67±0.00a* 19.00±0.00c* 13.04±0.06b*
Raw Gonder type mung bean 1.07±0.00e 2.07±0.03b 1.81±0.01a** 20.67±0.58c** 11.34±0.27b**
North showa type mung bean 1.06±0.00e* 2.04±0.02b 1.88±0.01a*** 12.00±0.00c*** 20.47±0.09b***
Bale type mung bean 1.00±0.01b 1.84±0.01a 1.85±0.01b* 17.67±0.58c* 12.67±0.49a*
Dehulled Gonder type mung bean 1.01±0.00b 1.77±0.01a 1.73±0.01b** 19.33±1.15c** 11.16±0.63a**
North showa type mung bean 1.00±0.00b* 1.90±0.02a 1.84±0.01b*** 14.33±0.58c*** 15.31±0.44a***
Bale type mung bean 1.01±0.00c 2.55±0.04d 1.88±0.01e* 8.67±0.58a* 31.79±2.34e*
Dry roasted Gonder type mung bean 1.00±0.00c 2.54±0.05d 1.77±0.01e** 7.33±0.58a** 37.65±2.83e**
North showa type mung bean 1.02±0.00c* 2.58±0.00d 1.98±0.00e*** 7.33±0.58a*** 37.55±2.75e***
Bale type mung bean 1.01±0.00d 2.10±0.01c 1.86±0.01c* 15.33±0.58d* 21.85±0.75c*
Germinated Gonder type mung bean 1.02±0.00d 2.10±0.01c 1.80±0.00c** 19.00±0.00d** 13.72±0.06c**
North showa type mung bean 1.03±0.00d* 2.20±0.02c 1.88±0.01c*** 19.33±0.58d*** 15.20±0.39c***
Bale type mung bean 1.02±0.00d 2.04±0.02c 1.97±0.01d* 11.00±0.00b* 22.42±0.11d*
Soaked Gonder type mung bean 1.01±0.00d 2.22±0.02c 1.73±0.00d** 19.67±0.58b** 12.48±0.40d**
North showa type mung bean 1.03±0.00d* 2.04±0.00c 1.90±0.01d*** 13.67±1.15b*** 20.65±1.54d***
Bale type mung bean 0.93±0.00a 3.22±0.02e 1.80±0.01b* 6.00±0.00a* 51.89±0.19f*
Boiled Gonder type mung bean 0.93±0.00a 3.14±0.02e 1.78±0.01b** 11.67±1.15a** 26.98±2.71f**
North showa type mung bean 0.94±0.00a* 3.16±0.01e 1.84±0.01b*** 6.00±0.00a*** 52.61±0.19f***
All values are the means of triplicates ± standard deviation n=3
Means with the same superscript letters within a column are not significantly different at (P>0.05)
a, b, c, d, e, f are superscript to show significance difference between means a<b<c<d<e<f
Means with superscript * indicated there is significant different between origin of mung bean (p<0.05)

4.2.1 Bulk density

The bulk density of raw and processed mung bean flour samples collected from Bale, Gonder
and North showa were ranged from 0.93% to 1.08% (Table 3). The highest bulk density
observed for raw Bale type mung bean flour and lowest bulk density observed for boiled Bale
type and Gonder type mung bean flour sample. In addition, germination and soaking, dry
roasting, dehulling, and boiling had decreased the bulk density of mung bean flour samples as
compared to the raw mung bean flour sample significantly (P < 0.05). Furthermore, there were
significant difference) among the bulk density of the North showa type, Bale type and Gonder
type mung bean flour sample (p<0.05). Consequently there was no significant difference
between the bulk density of Bale type and Gonder type mung bean flour sample (p>0.05). The
interaction of processing methods and origin of mung bean had significant effect on the bulk

34
density of mung bean flour sample (p<0.05). The result of this study disclose that all the five
processing methods (soaking, dehulling, boiling, germination and dry roasting) decreased the
bulk density of mung bean flour samples as compared to the raw mung bean flour sample. Bulk
density is influenced by the structure of the starch polymers and the loose structure of the starch
polymers by processing methods could result in low bulk density. The advantage of flour that has
high bulk density is that it does not take up too much space when distributing and decreases
packaging costs (Karuna et al., 1996).

4.2.2 Solubility index

The solubility index of raw and processed mung bean flour samples were ranged from 11.16% to
52.61% (Table 3). The maximum solubility index observed for boiled North showa type mung
bean flour and minimum solubility index observed for dehulled Gonder type mung bean flour.
Moreover, boiling, dry roasting, soaking and germination had significantly increased the
solubility index of mung bean flour sample as compared to the raw mung bean flour sample (P <
0.05). Furthermore, dehulling had significantly decreased the solubility index of mung bean flour
sample as compared to the raw mung bean flour sample (P < 0.05). Consequently there were
significant difference among the solubility index of the North showa type, Bale type and Gonder
type Mung bean flour samples (p<0.05). The interaction of processing methods and origin of
mung bean had significant effect on the solubility index of mung bean flour sample (p<0.05).
The result of the study shows that boiling, dry roasting, soaking and germination increased the
solubility index of mung bean flour samples as compared to raw mung bean flour sample. As a
direct result of flour swelling, there is a parallel increase in the solubility of flour. High solubility
implies high leaching. The high water solubility of any sample analyzed may be attributed to the
degree of swelling power and swelling power and solubility of the flour provide evidence of non-
covalent bonding between molecules within the flour (Onitilo et al., 2007).But dehulling
decreased the solubility index of mung bean flour sample as compared to raw mung bean flour
sample and it is probably due to the removal of seeds cover.

4.2.3 Swelling power

The swelling power of raw and processed mung bean flour samples were ranged from 7.33% to
20.67% (Table 3). The maximum swelling power observed for raw North showa type mung bean

35
flour and minimum swelling power observed for boiled Bale and North showa type mung bean
flour. In addition, germination had significantly increased the swelling power of mung bean flour
sample as compared to the raw mung bean flour sample (P < 0.05). Moreover, boiling, dry
roasting, and soaking had significantly decreased the swelling power of mung bean flour sample
as compared to the raw mung bean flour sample (P < 0.05). Furthermore, there was no
significant difference between the swelling power of dehulling mung bean flour sample and raw
mung bean flower sample(p>0.05). Consequently there were significant difference among the
swelling power of the North showa type, Bale type and Gonder type mung bean flour samples
(p<0.05). The interaction of processing methods and origin of mung bean had significant effect
on the swelling power of mung bean flour sample (p<0.05). The result of the study disclose that
boiling, dry roasting, and soaking decreased the swelling power of mung bean flour samples as
compared to raw mung bean flour sample. But germination increased the solubility index of
mung bean flour sample as compared to raw mung bean flour sample. The swelling power of
flour samples is often related to their protein and starch contents (Woolfe, 1992). A higher
protein content in flour may cause the starch granules to be embedded within a stiff protein
matrix, which subsequently limits the access of the starch to water and restricts the swelling
power. In addition to protein content, a higher concentration of phosphorous may increase
hydration and swelling power by weakening the extent of bonding within the crystalline domain
(Singh et al., 2003).Furthermore, the amylopectin is primarily responsible for granule swelling,
thus higher amylose content would reduce the swelling factor of starch (Tester and Morrison,
1990).The variation in the swelling power indicates the degree of exposure of the internal
structure of the starch present in the flour to the action of water (Ruales,1993). The result of this
study similar with previous study was conducted by Pangastuti et al. (2013).

4.2.4 Water Absorption Capacity

The water absorption capacity of raw and processed mung bean flour samples were ranged from
1.77% to 3.22% (Table 3). The maximum water absorption capacity observed for boiled Bale
type mung bean flour and minimum water absorption capacity observed for dehulled Gonder
type mung bean flour. Furthermore, boiling, dry roasting, and germination had significantly
increased the water absorption capacity of mung bean flour sample as compared to the raw mung
bean flour sample (P < 0.05). An increase in water absorption capacity by germination could be

36
attributed to the breakdown of polysaccharide and an increase in protein content, which probably
increased the sites for interaction of water molecules Elkhalifa and Bernhardt (2010).Moreover,
dehulling had significantly decreased the water absorption capacity of mung bean flour sample
as compared to the raw mung bean flour sample (P < 0.05). Consequently there were no
significant differences among the water absorption capacity of the North showa type, Bale type
and Gonder type mung bean flour sample(p>0.05). The interaction of processing methods and
origin of mung bean had significant effect on water absorption capacity of mung bean flour
sample (p<0.05).

4.2.5 Oil Absorption Capacity

The oil absorption capacity of raw and processed mung bean flour samples were ranged from
1.67% to 1.98% (Table 3). The maximum oil absorption capacity observed for dry roasted North
showa type mung bean flour and minimum oil absorption capacity observed for raw Bale type
mung bean flour sample. Moreover, dry roasting, soaking, germination, dehulling, and boiling
had significantly increased the oil absorption capacity of mung bean flour samples as compared
to the raw mung bean flour sample (P < 0.05). Consequently there were significant difference
among the oil absorption capacity of the North showa type, Gonder type and Bale type mung
bean flour samples (p<0.05). The interaction of processing methods and origin of mung bean had
significant effect on the oil absorption capacity of mung bean flour sample (p<0.05). The result
of this study show that all the five processing methods (soaking, dehulling, boiling, germination
and dry roasting) increased the oil absorption capacity of mung bean flour samples as compared
to the raw mung bean flour sample. Earlier studies also showed that the oil absorption capacity
of sorghum and brown rice flour increased as the germination time progressed Chinma et al.
(2015). Oil absorption capacity is an important property in food formulations because fats
improve the flavor and mouth feel of foods (Plaami, 1997).

4.3 Determination of Antinutritional Factors of Mung bean flour

4.3.1 Phytate
The phytate content of raw and processed mung bean flour samples collected from Bale, Gonder
and North showa ranged from 133.86% to 190.75% (Table 4). The maximum phytate content

37
observed for dehulled Bale type mung bean flour and minimum phytate content observed for raw
Gonder type mung bean flour. Furthermore, soaking, dehulling, boiling, and germination had
significantly decreased the phytate content of mung bean flour sample as compared to the raw
mung bean flour sample (P < 0.05). Moreover, dry roasting had significantly increased the
phytate content of mung bean flour sample as compared to the raw mung bean flour sample (P <
0.05). Consequently there were significant difference among the phytate content of the Bale type,
North showa type and Gonder type mung bean flour sample (p<0.05). The interaction of
processing methods and origin of mung bean had significant effect on the phytate content of
mung bean flour sample (p<0.05). The result of this study reveal that soaking, dehulling, boiling
and germination decreased the phytate content of mung bean flour sample as compared to the
raw mung bean flour sample. But dry roasting increased the phytate content of mung bean flour
sample as compared to the raw mung bean flour sample. The differences in the loss of phytic
acid contents during processing could probably be explained on the basis that phytases activity at
a temperature of 40–55C0 may degraded inositol hexaphosphate to the pentaphosphate or lower
molecular weight forms (De Boland et al.,1975). Phytic acid content decreased because insoluble
complexes between phytate and other components were formed during processing (Kumar et al.,
1978).

4.3.2 Tannin
The Tannin content of raw and processed mung bean flour samples were ranged from 13.69% to
23.71% (Table 4). The maximum tannin content observed for soaked Gonder type mung bean
flour and minimum tannin content observed for germinated Gonder type mung bean flour. In
addition, germination and dehulling had significantly decreased the tannin content of mung bean
flour sample as compared to the raw mung bean flour sample (P < 0.05). Furthermore, dry
roasting and soaking significantly increased the tannin content of mung bean flour sample as
compared to the raw mung bean flour sample (P < 0.05). Moreover, there were no significant
difference between the tannin content of boiled and raw mung bean flour sample (P >0.05).
Consequently there were significant difference among the tannin content of the Bale type,
Gonder type and North showa type mung bean flour sample(p<0.05). The interaction of
processing methods and origin of mung bean had significant effect on the tannin content of mung
bean flour sample (p<0.05). The result of this study disclose that soaking, dehulling, boiling and

38
germination decreased the tannin content of mung bean flour sample as compared to the raw
mung bean flour sample. But dry roasting and soaking increased the tannin content of mung bean
flour sample as compared to the raw mung bean flour sample. The decreased during processing
might be due to thermal degradation and denaturation of the antinutrients as well as the
formation of insoluble complexes and leaching out of hydrolysable tannin in the boiling water
(Osman,2004).

Table 4. Antinutritional contents of raw and processed mung bean flour

Phytate
Method of content(µg/100 Tannin(mg/10
processing Origin of mung bean gm) 0gm)
Bale type mung bean 182.59±0.08e* 19.29±0.18c*
Raw Gonder type mung bean 133.86±0.91e** 17.25±0.15c**
North showa type mung bean 176.14±0.01e*** 18.71±0.31c***
Bale type mung bean 190.75±0.02b* 18.41±0.05b*
Dehulled Gonder type mung bean 134.51±0.70b** 17.15±0.08b**
North showa type mung bean 148.04±0.22b*** 17.98±0.09b***
Bale type mung bean 179.46±0.09f* 20.43±0.20e*
Dry roasting Gonder type mung bean 184.23±0.31f** 20.60±0.19e**
North showa type mung bean 183.94±0.82f*** 22.22±0.07e***
Bale type mung bean 181.54±0.01d* 17.95±0.50a*
Germinating Gonder type mung bean 134.31±0.91d** 13.69±0.10a**
North showa type mung bean 169.06±0.28d*** 12.93±0.91a***
Bale type mung bean 138.05±0.30a* 17.30±0.29d*
Soaked Gonder type mung bean 145.72±0.16a** 23.71±0.13d**
North showa type mung bean 145.43±0.39a*** 19.25±0.33d***
Bale type mung bean 163.23±0.36c* 19.77±0.12c*
Boiled Gonder type mung bean 134.31±0.91c** 17.93±0.08c**
North showa type mung bean 183.06±0.69c*** 17.27±0.08c***
All values are the means of triplicates ± standard deviation n=3
Means with the same superscript letters within a column are not significantly different at (P>0.05)
a, b, c, d, e, f are superscript to show significance difference between means a<b<c<d<e<f
Values are expressed in µg/100gm and mg/100g of dry weight basis
Means with superscript * indicated there is significant different between origin of mung bean (p<0.05)

4.4 Antioxidant activities of mung bean flour

4.4.1 Yield of extract

The percentage of yields of extract raw and processed mung bean flour samples collected from
Bale, Gonder and North showa by methanol were ranged from 13.4% to 2.8% (Table 5). From

39
the yield of extract the free radical scavenging capacity by DPPH, total phenol and flavonoid was
determined.

Table 5 Percentage of yields of extract by methanol from raw and processed mung bean
Sample type Raw Dehulled Soaked Germinated Boiled Dry roasted
Bale type 5 5 4.8 11.6 3 4.8
Gonder type 5.4 2.8 9 12 2.8 5.4
North showa type 7.6 8.6 4.4 11.8 3.8 13.4

4.4.2 Determination of radicals scavenging activity of mung bean by DPPH

Radical scavenging activity of the sample extracts was measured by determining the inhibition
rate of DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical. DPPH is stable free radical at room
temperature and accepts an electron / hydrogen radical to become a stable diamagnetic molecule
(J.M, 2004). The reduction capability of DPPH radical is determined by the decrease in its
absorbance at 5l7 nm, induced by antioxidants. The decreased in absorbance of DPPH radical is
caused by antioxidants, because of the reaction between antioxidant molecules and radicals,
progresses, which results in the scavenging of the radical by hydrogen donation. It is visually
noticeable as a change in color from purple to yellow. Hence, DPPH is usually used as a
substrate to evaluate the antioxidative activity (R.Edamatsu, 1989).The ability of mung bean
extracts to quench reactive species by hydrogen donation was determined by the DPPH radical
scavenging activity. The antioxidant of the sample can react with DPPH free radical, a violet
color is converted in to yellow color of α, α-diphenil-β-picryllhydrazine. The discoloration of the
reaction mixture can be quantified by measuring the absorbance at 517nm, which indicates the
radical scavenging ability of the antioxidant. The concentration antioxidant of the sample at IC50
was determined from the graph of %inhibition vs concentration.

40
Fig7: Percentage inhibition verse concentration of samples and ascorbic acid at
IC50 by DPPH

At the concentration 50mg/ml, the scavenging effect of ascorbic acid and mung bean extract
which is collected from Bale, Gonder and North showa on the DPPH radical scavenging
decreased in the order of ascorbic acid>boiled >raw>soaked>germinated>dehulled. From the
above figure the concentration of the raw and processed mung bean that scavenged free radical
ranged from 4.67mg/ml to 31.41mg/ml, which means the minimum value was for ascorbic acid
that had the largest antioxidizing activity and the maximum value was for dehulled Gonder type
mung bean which had less oxidizing activity. The reduction of antioxidant of the dehulled
Gonder and North showa type mung bean sample was due to the removal of the seed cover of
mung bean. As concentration increased from 4.67mg/ml to 31.41 mg/ml, the antioxidant
activities decreased.

41
Table 6 Concentration of ascorbic acid and samples at IC50
Variety Ascorbic Raw Dehulled Soaked Germinated Boiled Dry roasted
acid
Bale type Mung 4.67 13.26 15.61 15.17 9.99 11.11 8.35
bean
Gonder type Mung 4.67 14.84 31.41 6.5 15.44 4.99 14.78
bean
North showa type 4.67 16.68 30.86 15.7 28.39 13.12 19.39
mung bean

4.4.3 Determination of Total Phenol

The total phenol content of raw and processed mung bean flour samples were ranged from 14.28
to 125.72 mg GAE/g extract (Table 6). The maximum total phenol content observed for
germinated Bale type mung bean flour and minimum total phenol content observed for dry
roasted North showa type mung bean flour. Germination had significantly increased the total
phenol content of mung bean flour sample as compared to the raw mung bean flour sample (P <
0.05). Furthermore, dry roasting, boiling, soaking and dehulling had significantly decreased the
total phenol content of mung bean flour sample as compared to the raw mung bean flour sample
(P < 0.05). Moreover, there were significant difference among the total phenol content of the
Bale type, Gonder type and North showa type mung bean flour sample(p<0.05). Consequently,
there was no significant difference between Gonder type and North showa type mung bean flour
sample (p>0.05). The interaction of processing methods and origin of mung bean had significant
effect on the total phenol content of mung bean flour sample (p<0.05). The result of this study
reveal that germination increased the total phenol content of mung bean flour sample as
compared to the raw mung bean flour sample. But dry roasting, soaking, boiling and dehulling
decreased the total phenol content of mung bean flour sample as compared to the raw mung bean
flour sample. This shows an increased in phenolic content during germination. Germination
process causes release of microbial enzyme which in turn produces more freely available form of
plant chemicals like flavonoid, alkaloid and phenyl propanoids (Messens and Vuyst 2002).

42
4.4.4 Determination of total flavonoid
The total flavonoid content of raw and processed mung bean flour samples were ranged from
42.38 to 190.99 mg DE/g. (Table 6). The maximum total flavonoid content observed for
germinated Bale type mung bean flour and minimum total flavonoid content observed for
dehulled Gonder type mung bean flour. Moreover, germination and dry roasting had significantly
increased the total flavonoid content of mung bean flour sample as compared to the raw mung
bean flour sample (P < 0.05). Furthermore, dehulling, boiling, and soaking had significantly
decreased the total flavonoid content of mung bean flour sample as compared to the raw mung
bean flour sample (P < 0.05). Consequently, there were significant difference among the total
flavonoid content of the North showa type, Bale type and Gonder type mung bean flour sample
(p<0.05). The interaction of processing methods and origin of mung bean had significant effect
on the flavonoid content of mung bean flour sample (p<0.05). The result of this study reveal that
germination and dry roasting increased the total flavonoid content of mung bean flour sample as
compared to the raw mung bean flour sample. But soaking, boiling and dehulling decreased the
total flavonoid content of mung bean flour sample as compared to the raw mung bean flour
sample. Germination increased the flavonoid content might be due to microbial enzymes, such as
glucosidase, amylase, cellulase, tannase, esterase, invertase or lipase produced during
germination can hydrolyse glucosides, and break down plant cell walls or starch. These enzymes
play a role in disintegrating the plant cell wall matrix and consequently facilitating the flavonoids
extraction (Hur et al., 2014). Another mechanism is along germination the β- glucosidase of
microbial origin could also be used to hydrolyze the phenolic and flavonoids.

43
Table 7 Total phenol and flavonoid contents of raw and processed mung bean
Method of Total Phenol(mg Total Flavonoid(mg
processing Variety GAE/g extract) DE/g)
Bale type mung bean 25.71±0.87c* 66.39±0.73d*
Raw Gonder type mung bean 45.14±0.40c, 77.72±0.62d**
North showa type mung bean 37.27±0.46c 87.29±0.67d***
Bale type mung bean 23.28±0.97b* 58.45±0.91a*
Dehulled Gonder type mung bean 19.04±0.68b 42.38±0.79a**
North showa type mung bean 43.85±0.15b 44.19±0.56a***
Bale type mung bean 36.88±1.06a* 114.24±0.58e*
Dry roasting Gonder type mung bean 23.62±0.36a 97.23±0.18e**
North showa type mung bean 14.28±0.92a 151.10±0.17e***
Bale type mung bean 125.72±13.26d* 190.99±0.80f*
Germinating Gonder type mung bean 54.98±0.60d 143.94±0.65f**
North showa type mung bean 65.23±0.53d 173.40±0.65f***
Bale type mung bean 26.46±2.29b* 71.53±0.58c*
Soaked Gonder type mung bean 39.69±0.97b 78.32±0.70c**
North showa type mung bean 29.75±1.13b 66.08±0.29c***
Bale type mung bean 18.37±0.63b* 49.51±0.09b*
Boiled Gonder type mung bean 35.60±0.47b 44.92±0.37b**
North showa type mung bean 32.99±0.54b 60.79±0.34b***
All values are the means of triplicates ± standard deviation n=3
Means with the same superscript letters within a column are not significantly different at (P>0.05)
a, b, c, d, e, f are superscript to show significance difference between means a<b<c<d<e<f
Means with superscript * indicated there is significant different between origin of mung bean (p<0.05)

44
 CHAPTER FIVE
5. CONCLUSION AND RECOMMENDATION

5.1. Conclusion

Mung bean is one of the exported commodity in Ethiopian Commodity Exchange and consumed
pules in Ethiopia. This study attempted to investigate the effect of five processing methods
(boiling, dehulling, dry roasting, germination and soaking) on nutritional composition, functional
properties, antinutritional factors, and antioxidant activities of raw and processed mung bean
flour collected from Bale, Gonder and North showa. From the results of the present study it was
understood that mung bean contains adequate amount of carbohydrate, crude fiber, and crude
protein. Furthermore, the results of this study also showed that mung bean contains low levels of
antinutrients (phytate and tannin) when compared to other pulses and cereals. Moreover, there
were further reductions of the antinutritional factors during processing. In addition, it was
observed in the study that functional properties of mung bean flour was remarkably higher.
Moreover, high water absorption capacity and swelling power than other pulses flour was
recorded. However, the solubility index and oil absorption capacity was in the range.
Consequently, evaluation of the five processing methods in terms of antinutrients reduction and
nutrients enrichment indicated that all the five processing methods were found to be effective in
the reduction of antinutrients but effect of germination was found to be highest in the reduction
of antinutritional factors. In relation to nutritional profile, the low protein content of mung bean
flour sample was observed to be increased by germination and total phenol and flavonoid
contents also increased by germination and dehulling. Germination of mung bean flour was a
more acceptable process as it was inexpensive, fuel efficient method and environmentally
friendly by which people can obtain good quality food and this process can only be performed at
their own homes. However, boiling was found to decreased most of the functional properties of
mung bean flour. Germination and dry roasting increased oil absorption and reduced most of
antinutritional factor of mung bean samples. Therefore among the five traditional applied
processing methods germination and dry roasting was recommended process. Finally among the
three types of export mung bean varieties (Bale type mung bean, Gonder type mung bean and
North Showa type mung bean) in terms of proximity composition, functional properties,

45
antinutritional factor and antioxidant activities Bale type mung bean was better than Gonder type
and North showa type mung bean.

5.2 Recommendations

Due to the low attention given to mung bean in Ethiopia and limitation of mung bean in specific
regions and yet no research on it, mung bean cannot known as functional food in Ethiopia

 Encourage public awareness on the importance of mung bean and community based
management of these pulse through avoiding antinutritional factors by processing.
 Encourage research on this plant, especially for its potential as a nutraceutical, or
pharmaceutical product for use in the treatment of obesity, obesity-related dyslipidemia
diabetes, hyperlipidemia and hypercholesterolemia.
 Encourage farmers to cultivate and conserve mung bean that grow in their farmlands to
be used in times of drought.
 Analysis should be done for the vitamins and minerals content of mung bean.
 Investigation on more specific properties, such as baking property, of these composite
flours should be undertaken in the future to fully investigate their specific application.
 Functional property of mung bean is relatively higher than other pulses, therefore it is a
good input for product formulation with other crop.
 The starch characteristic of mung bean should be studied to know how much
amylose/amylopectin and gelatinization properties.
 Sensory evaluation of the product should be developed.

46
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 Appendix-1

1. ANOVA and Duncan table of proximity analysis

1.1 ANOVA table of moisture content
Source Sum of df Mean F Sig.
Squares square
Corrected Model 144.327a 17 8.49 98.80 0.000
Intercept 2188.86 1 2188.86 25473.80 0.000
Processing method 129.91 5 25.98 302.374 0.000
Origin 0.164 2 0.082 .957 0.394
Processing method * Origin 14.25 10 1.425 16.588 0.000
Error 3.09 36 0.086
Total 2336.28 54
Corrected Total 147.42 53

1.2 Duncan table of moisture content

Subset
Method of processing N 1 2 3 4
Duncana,b dry roasted 9 4.1333
dehulled 9 5.2889
germinated 9 5.3333
soaked 9 7.0444
boiled 9 8.0667
raw 9 8.3333
Sig. 1.000 .750 1.000 .062
Means for groups in homogeneous subsets are displayed.
Based on observed means. The error term is Mean Square (Error) = .086.
a. Uses Harmonic Mean Sample Size = 9.000.
b. Alpha = .05.

1.3 ANOVA table of Crude protein

Source Sum of df Mean F Sig.
Squares square
Corrected Model 42.419a 17 2.495 13.845 0.000
Intercept 47746.976 1 47746.976 264928.456 0.000
Processing method 37.786 5 7.557 41.932 0.000
Origin 1.153 2 0.577 3.199 0.053
Processing method * Origin 3.480 10 0.348 1.931 0.073
Error 6.488 36 0.180
Total 47795.883 54
Corrected Total 48.907 53

53
1.4 Duncan table of Crude protein
Subset
Method of processing N 1 2 3 4
Duncana,b raw 9 28.0422
soaked 9 29.4011
dry roasted 9 30.0200
boiled 9 30.0389
germinated 9 30.3867 30.3867
dehulled 9 30.5244
Sig. 1.000 1.000 .091 .496
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.180.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

1.5 Duncan table Export type mung bean origin of Crude protein

Subset
Export type mung bean Origin N 1 2
Duncana,b Bale type mung bean 18 29.5600
Gonder type mung bean 18 29.7289 29.7289
North showa type mung bean 18 29.9178
Sig. .240 .190
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.180.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05.

1.6 ANOVA table of Crude fat

Source Sum of df Mean F Sig.

Squares square
Corrected Model 2.856a 17 0.168 36.294 0.000
Intercept 111.227 1 111.227 24025.000 0.000
Processing method 0.634 5 0.127 27.400 0.000
Origin 0.454 2 0.227 49.000 0.000
Processing method * Origin 1.769 10 0.177 38.200 0.000
Error 0.167 36 0.005
Total 114.250 54
Corrected Total 3.023 53

54
1.7 Duncan table method of processing of Crude fat

Subset
Method of processing N 1 2 3
Duncana,b dehulled 9 1.3333
soaked 9 1.3333
boiled 9 1.3333
raw 9 1.5000
germinated 9 1.5000
dry roasted 9 1.6111
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.005.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

1.8 Duncan table Export type mung bean variety of Crude fat

Subset
Export type mung bean Origin N 1 2
Duncana,b Bale type mung bean 18 1.3056
Gonder type mung bean 18 1.5000
North showa type mung bean 18 1.5000
Sig. 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.005.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05.

1.9 ANOVA table of Total ash

Source Sum of df Mean F Sig.

Squares square
Corrected Model 9.659a 17 0.568 12.784 0.000
Intercept 517.701 1 517.701 11648.267 0.000
Processing method 9.197 5 1.839 41.387 0.000
Origin 0.077 2 0.039 0.867 0.429
Processing method * Origin 0.385 10 0.039 0.867 0.571
Error 1.600 36 0.044
Total 528.960 54
Corrected Total 11.259 53

55
1.10 Duncan table of method of processing of total ash

Subset
Method of processing N 1 2 3 4
Duncana,b boiled 9 2.2222
dehulled 9 3.0667
soaked 9 3.1556 3.1556
raw 9 3.2889 3.2889
dry roasted 9 3.3778
germinated 9 3.4667
Sig. 1.000 .377 .188 .099
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.044.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

1.11 ANOVA table of Crude fiber

Source Sum of df Mean F Sig.

Squares square
Corrected Model 222.590a 17 13.094 316.368 0.000
Intercept 310.704 1 310.704 7507.280 0.000
Processing method 198.460 5 39.692 959.045 0.000
Origin 5.705 2 2.853 68.924 0.000
Processing method * Origin 18.425 10 1.842 44.518 0.000
Error 1.490 36 0.041
Total 534.784 54
Corrected Total 224.080 53

1.12 Duncan table of method of processing of Crude fiber

Subset
Method of processing N 1 2 3 4
Duncana,b soaked 9 1.2756
dehulled 9 1.4567 1.4567
boiled 9 1.4656 1.4656
germinated 9 1.5056
dry roasted 9 2.0356
raw 9 6.6533
Sig. .068 .635 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.041.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

56
1.13 Duncan table of Export type mung bean variety Crude fiber

Subset
Export type mung bean Origin N 1 2
Duncana,b Bale type mung bean 18 2.1633
Gonder type mung bean 18 2.1744
North showa type mung bean 18 2.8583
Sig. .871 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.041.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05.

1.14 ANOVA table of Utilizable carbohydrate

Source Sum of df Mean F Sig.

Squares square
Corrected Model 307.168a 17 18.069 44.792 0.000
Intercept 175246.557 1 175246.557 434433.761 0.000
Processing method 266.281 5 53.256 132.021 0.000
Origin 6.228 2 3.114 7.720 0.002
Processing method * Origin 34.659 10 3.466 8.592 0.000
Error 14.522 36 0.403
Total 175568.247 54
Corrected Total 321.690 53
1.15 Duncan table of method of processing of Utilizable carbohydrate

Subset
Method of processing N 1 2 3 4
Duncana,b raw 9 52.1822
boiled 9 56.8733
soaked 9 57.7900
germinated 9 57.8078
dehulled 9 58.3300 58.3300
dry roasted 9 58.8222
Sig. 1.000 1.000 .096 .109
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.403.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

57
1.16 Duncan table of Export type mung bean variety Utilizable carbohydrate

Subset
Export type mung bean Origin N 1 2
Duncana,b North showa type mung bean 18 56.4906
Bale type mung bean 18 57.1578
Gonder type mung bean 18 57.2544
Sig. 1.000 .651
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.403.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05.

58
Appendix-2

2. ANOVA and Duncan table of functional properties

2.1 ANOVA table of Bulk density
Source Sum of df Mean F Sig.
Squares square
Corrected Model 0.090a 17 0.005 756.174 0.000
Intercept 54.888 1 54.888 7799819.379 0.000
Processing method 0.088 5 0.018 2492.726 0.000
Origin 0.000 2 0.000 28.337 0.000
Processing method * Origin 0.002 10 0.000 33.465 0.000
Error 7.04E-006 36 7.04E-006
Total 54.978 54
Corrected Total 0.091 53

2.2 Duncan table of method of processing of Bulk density

Subset
Method of processing N 1 2 3 4 5
Duncana,b boiled 9 .9318
dehulled 9 1.0022
dry roasted 9 1.0111
soaked 9 1.0164
germinated 9 1.0187
raw 9 1.0689
Sig. 1.000 1.000 1.000 .084 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
7.04E-006.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

2.3 Duncan table of Export type mung bean variety of Bulk density
Subset
Export type mung bean Origin N 1 2
Duncana,b Bale type mung bean 18 1.0054
Gonder type mung bean 18 1.0072
North showa type mung
18 1.0119
bean
Sig. .052 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
7.04E-006.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05.

59
2.4 ANOVA table of Solubility
Source Sum of df Mean F Sig.
Squares square
Corrected Model 9093.415a 17 534.907 294.628 0.000
Intercept 30645.383 1 30645.383 16879.549 0.000
Processing method 7299.149 5 1459.830 804.078 0.000
Origin 673.263 2 336.632 185.417 0.000
Processing method * Origin 1121.004 10 112.100 61.745 0.000
Error 65.359 36 1.816
Total 39804.158 54
Corrected Total 9158.775 53

2.5 Duncan table of method of processing of Solubility

Subset
Method of processing N 1 2 3 4 5 6
Duncana,b dehulled 9 13.0500
raw 9 14.9511
germinated 9 16.9200
soaked 9 18.5189
dry roasted 9 35.6667
boiled 9 43.8278
Sig. 1.000 1.000 1.000 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
1.816.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

2.6 Duncan table of Export type mung bean variety of Solubility

Subset
Export type mung bean Origin N 1 2 3
Duncana,b Gonder type mung bean 18 18.8906
Bale type mung bean 18 25.6106
North showa type mung
18 26.9661
bean
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
1.816.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

60
2.7 ANOVA table of swelling power

Source Sum of Squares df Mean square F Sig.

Corrected Model 1385.333a 17 81.490 209.546 0.000
Intercept 10250.667 1 10250.667 26358.857 0.000
Processing method 1004.000 5 200.800 516.343 0.000
Origin 175.000 2 87.500 225.000 0.000
Processing method * Origin 206.333 10 20.633 53.057 0.000
Error 14.000 36 0.389
Total 11650.000 54
Corrected Total 1399.333 53

2.8 Duncan table of method of processing of swelling power

Subset
Method of processing N 1 2 3 4
Duncana,b dry roasted 9 7.7778
boiled 9 7.8889
soaked 9 14.7778
dehulled 9 17.1111
raw 9 17.2222
germinated 9 17.8889
Sig. .708 1.000 .708 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.389.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

2.9 Duncan table of Export type mung bean variety of swelling power

Subset
Export type mung bean Origin N 1 2 3
Duncana,b North showa type mung
18 12.1111
bean
Bale type mung bean 18 12.9444
Gonder type mung bean 18 16.2778
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.389.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

61
2.10 ANOVA table of Water Absorption Capacity

Source Sum of df Mean F Sig.

Squares square
Corrected Model 10.843a 17 0.638 162.312 0.000
Intercept 286.719 1 286.719 72963.360 0.000
Processing method 10.694 5 2.139 544.263 0.000
Origin 0.013 2 0.007 1.664 0.204
Processing method * Origin 0.136 10 0.014 3.466 0.003
Error 0.141 36 0.004
Total 297.703 54
Corrected Total 10.985 53

2.11 Duncan table of method of processing Water Absorption Capacity

Subset
Method of processing N 1 2 3 4 5
Duncana,b dehulled 9 1.8400
raw 9 2.0200
soaked 9 2.1011
germinated 9 2.1344
dry roasted 9 2.5544
boiled 9 3.1756
Sig. 1.000 1.000 .267 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.004.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

2.12 ANOVA table of Oil Absorption Capacity

Source Sum of df Mean F Sig.

Squares square
Corrected Model 0.327a 17 0.019 259.300 0.000
Intercept 180.914 1 180.914 2442336.400 0.000
Processing method 0.064 5 0.013 172.600 0.000
Origin 0.124 2 0.062 834.100 0.000
Processing method * Origin 0.139 10 0.014 187.690 0.000
Error 0.003 36 0.000074
Total 181.243 54
Corrected Total 0.329 53

62
2.13 Duncan table of method of processing of Oil Absorption Capacity

Subset
Method of processing N 1 2 3 4 5
Duncana,b raw 9 1.7844
dehulled 9 1.8033
boiled 9 1.8056
germinated 9 1.8467
soaked 9 1.8656
dry roasted 9 1.8767
Sig. 1.000 .587 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
7.41E-005.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

2.14 Duncan table of Export type mung bean variety of Oil Absorption Capacity

Subset
Export type mung bean Origin N 1 2 3
Duncana,b Gonder type mung bean 18 1.7689
Bale type mung bean 18 1.8367
North showa type mung bean 18 1.8856
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
7.41E-005.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

63
Appendix-3

3. Standard Curve, ANOVA and Duncan table of Antinutritional factor

3.1 Standard Curve of phytate

3.2 ANOVA table of phytate

Source Sum of Squares df Mean square F Sig.
Corrected Model 23983.973a 17 1410.822 5320.324 0.000
Intercept 1409620.697 1 1409620.697 5315794.380 0.000
Processing method 7251.338 5 1450.268 5469.077 0.000
Origin 8100.693 2 4050.346 15274.186 0.000
Processing method * Origin 8631.942 10 863.194 3255.176 0.000
Error 9.546 36 0.265
Total 1433614.216 54
Corrected Total 23993.520 53

64
3.3 Duncan table of method of processing of phytate
Subset
Method of processing N 1 2 3 4 5 6
Duncana,b soaked 9 143.0644
dehulled 9 157.7644
boiled 9 160.1978
germinated 9 161.6356
raw 9 164.1978
dry roasted 9 182.5456
Sig. 1.000 1.000 1.000 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.265.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.
3.4 Duncan table of Export type mung bean variety of phytate

Subset
Export type mung bean Origin N 1 2 3
Duncana,b Gonder type mung
18 144.4878
bean
North showa type
18 167.6117
mung bean
Bale type mung bean 18 172.6033
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.265.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

3.5 Standard Curve of Tannin

65
3.6 ANOVA table of Tannin
Source Sum of df Mean F Sig.
Squares square
Corrected Model 340.178a 17 20.010 228.386 0.000
Intercept 18352.964 1 18352.964 209468.871 0.000
Processing method 206.242 5 41.248 470.782 0.000
Origin 5.819 2 2.909 33.206 0.000
Processing method * Origin 128.117 10 12.812 146.224 0.000
Error 3.154 36 0.088
Total 18696.296 54
Corrected Total 343.332 53

3.7 Duncan table of method of processing of Tannin

Subset
Method of processing N 1 2 3 4 5
Duncana,b germinated 9 14.8556
dehulled 9 17.8456
boiled 9 18.3233
whole 9 18.4178
soaked 9 20.0889
dry roasted 9 21.0822
Sig. 1.000 1.000 .503 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.088.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

3.8 Duncan table of Export type mung bean variety of Tannin

Subset
Export type mung bean Origin N 1 2 3
Duncana,b North showa type mung bean 18 18.0589
Gonder type mung bean 18 18.3889
Bale type mung bean 18 18.8589
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.088.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

66
Appendix-4

4 Graph of percent of inhibition for three type Mung bean of DPPH, Standard curve,
ANOVA and Duncan table of Antioxidant Activity
4.1 Graph of percent of inhibition for three type Mung bean of DPPH

67
4.2 Standard curve of Total Phenol

68
4.3 ANOVA table of total Phenol
Source Sum of df Mean F Sig.
Squares square
Corrected Model 32752.587a 17 1926.623 183.038 0.000
Intercept 81165.770 1 81165.770 7711.10 0.000
Processing method 20778.126 5 4155.625 394.803 0.000
Origin 431.672 2 215.836 20.505 0.000
Processing method * Origin 11542.790 10 1154.279 109.662 0.000
Error 378.930 36 10.526
Total 114297.287 54
Corrected Total 33131.517 53

4.4 Duncan table of method of processing of total Phenol

Subset
Method of processing N 1 2 3 4
Duncana,b dry roasted 9 24.9267
dehulled 9 28.7222
boiled 9 28.9867
soaked 9 31.9656
raw 9 36.0411
germinated 9 81.9744
Sig. 1.000 .051 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
10.526.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

4.5 Duncan table of Export type mung bean variety of total Phenol

Subset
Export type mung bean Origin N 1 2
Duncana,b Gonder type mung bean 18 36.3467
North showa type mung bean 18 37.2261
Bale type mung bean 18 42.7356
Sig. .421 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
10.526.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

69
4.6 Standard curve of Flavonoid

4.7 ANOVA table of Flavonoid

Source Sum of df Mean square F Sig.

Squares
Corrected Model 108268.596a 17 6568.388 18567.731 0.000
Intercept 436572.392 1 436572.392 1272874.529 0.000
Processing method 98575.374 5 19715.075 57481.456 0.000
Origin 2519.479 2 1259.739 3672.908 0.000
Processing method * Origin 7167.743 10 716.774 2089.834 0.000
Error 12.347 36 0.343
Total 544847.335 54
Corrected Total 108274.943 53

4.8 Duncan table of method of processing of Flavonoid

Subset
Method of processing N 1 2 3 4 5 6
Duncana,b dehulled 9 48.3389
boiled 9 51.7411
soaked 9 71.9789
raw 9 77.1322
dry roasted 9 120.8556
germinated 9 169.4422
Sig. 1.000 1.000 1.000 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.343.
a. Uses Harmonic Mean Sample Size = 9.00
b. Alpha = .05.

70
4.9 Duncan table of Export type mung bean variety

Subset
Export type mung bean Origin N 1 2 3
Duncana,b Gonder type mung bean 18 80.7500
Bale type mung bean 18 91.8533
North showa type mung
18 97.1411
bean
Sig. 1.000 1.000 1.000
Means for groups in homogeneous subsets are displayed. Based on observed means. The error term is Mean Square (Error) =
.343.
a. Uses Harmonic Mean Sample Size = 18.000.
b. Alpha = .05

71