ARBAMINCH UNIVERSITY

SAWLA CAMPUS

ARBAMINCH INSTITUTE OF TECHNOLOGY

DEPARTMENT OF FOOD TECHNOLOGY AND PROCESS

ENGINEERING

A thesis submitted to the Department of Food Technology and

Process engineering Arbaminch University

In partial fulfillment of the requirements for the degree of bachelor

of sciences in Food Technology and Process engineering

Development and characterization of MORINGA fortified blended

food powder from peanut and soybean

NAME: DAWIT TADESSE

ID: RAMIT/568/11

JANUARY 2021

SAWLA, ETHIOPA
Acronyms

FBF Fortified Blended Food

CSB Corn Soy Blend

USAID

FAQR Food Aid Quality Review

WFP World Food Program

WHO World Health Organization

AOAC Association Of Official Analytical Chemists

OCS Organosulfur Compounds

MLP Moringa Leaf Powder

ANOVA Analysis Of Variance

GTP Growth And Transformation Plan

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 Catalog

Chapter one 1
1 Introduction 1
1.1 Background 1
1.1.1 Supplementary foods 2
1.2 Statement of the problem 3
1.3 Objectives 5
1.3.1 General objectives 5
1.3.2 Specific objective 5
1.4 Scope of the study 5
1.5 Significance of the study 5
CHAPTER TWO 7
2 Literature review 7
2.1 Malnutrition 7
2.2 Malnutrition in Ethiopia 7
2.3 Production and usage of soybean, peanut and MORINGA 8
2.3.1 Peanut 8
2.3.2 Soybean 9
2.3.3 MORINGA 10
Bio active compounds and their sources 14
2.4 Fortified blended foods 15
CHAPTER THREE 17
3 Materials and method 17
3.1 Description of the study area 17
3.2 Sample Collection and Transportation 17
3.3 Raw material preparation 17
3.3.1 Soybean flour preparation 17
3.3.2 Peanut flour preparation 18
3.3.3 MORINGA powder preparation 18
3.4 Functional properties of flours 19
3.4.1 Water Absorption Capacity (WAC) 19
3.5 Proximate Analysis 20
3.5.1 Moisture Content determination 21

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 3.5.2 Total Ash determination 21
3.5.3 Determination of Crude Protein 22
3.5.4 Crude Fat Determination 23
3.5.5 Crude Fiber determination 24
3.5.6 Determination of Total Carbohydrate 24
3.5.7 Calorific Value 25
3.6 Test for bioactive compounds 26
3.6.1 Condensed tannins 26
3.6.2 Phytic acid content 26
3.6.3 Total phenolic compounds 26
3.7 Production of experimental FBF 27
3.7.1 Formulation of experimental food blends 27
Ingredient blending ratio 27
4 Reference 29

Chapter one

1 Introduction

1.1 Background

Traditional complementary foods in the developing countries are known to be of low

nutritive value and are characterized by low protein, low energy density and high
bulk, because they are usually cereal–based. The protein content of cereals such as
maize and guinea corn, which is often used, is of poor quality, being low in lysine and
tryptophan amino acids which are indispensable for the growth of the young child.
For example maize porridge or bread has been implicated in the aetiology of
protein-energy malnutrition in children during the complementary period (Solomon,
M. (2005)). This nutritional deficiency can be corrected by several ways, one of
which is supplementation with grain legumes or oil seeds and fortification with
MORINGA stenopetala leaf powder (Solomon, M. 2005,Olushola, A.T.E. 2006)

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Soybean is an important source of high quality, inexpensive protein and oil. At 38%
soybean has the highest protein content of all food crops and is second only to peanut
in terms of oil content (18%) among food legumes. Compared to other protein-rich
foods such as meat, fish, and eggs, soybeans is by far the cheapest. It also has a
superior amino-acid profile compared to other sources of plant protein. The crops
main use is flour, protein products and animal feed (WHO 1998, Saskia, D.P. and
Martin, W.B. 2008).

Groundnut contains high quality edible oil, easily digestible protein and
carbohydrates. A nutritious peanut butter is prepared from groundnut. It is also a
significant source of resveratrol, a chemical compound that is reported to have a
number of beneficial health effects, such as anti-cancer, antiviral, neuro protective,
anti-aging, anti-inflammatory and life prolonging effects (Saskia, D.P. and Martin,
W.B. 2008).

MORINGA is a “miracle plant” that has almost all the minerals and vitamins that the
body needs for vibrant and good health. The leaves, pods and flowers of this plant
which are used as vegetable in many parts of the world have great nutritional value
(Fuglie, L.J. 2001). Almost all parts of the plant have therapeutic value. The leaves
are especially beneficial in the treatment of many ailments due to their various
medicinal properties and their rich iron content. MORINGA leaves and pods can be
an extremely valuable source of nutrients for people of all ages. The leaves can be
dried, made into powder (Olushola, A.T.E. 2006) and stored for use when needed.
Strategic use of such inexpensive high protein sources that complement the amino
acid pattern of cereal staple foods is highly recommended to upgrade the nutritional
status and prevent protein-energy malnutrition in the developing world.

The fortification of maize, soybean and peanut food formulations with MORINGA
oleifera leaf powder can dramatically improve their protein quality and micro nutrient
content, with little or no increase in the production cost (El-Adawy, T.A. 1997).

1.1.1 Supplementary foods

Supplementary foods are specially formulated foods, in ready-to-eat or in milled
form, which are modified in their energy density, protein, fat or micro-nutrient

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composition to help meet the nutritional requirements of specific populations.
Supplementary foods are not intended to be the only source of nutrients and are
different from complementary foods, in that the latter are intended for progressive
adaptation of infants 6 months of age and older to the food of the family.

They are also different from food supplements, which refer to vitamin and mineral
supplements in unit dose forms such as capsules, tablets, powders or solutions, where
national jurisdictions regulate these products as food. Supplementary foods have been
used to rehabilitate moderately malnourished persons or to prevent a deterioration of
nutritional status of those most at risk by meeting their additional needs, focusing
particularly on children 6–59 months of age, pregnant women and lactating mothers.
Examples of supplementary foods include fortified blended foods (FBF), which can
be used to prepare smooth, ready-to-eat porridges, and lipid-based nutrient
supplements.

1.2 Statement of the problem

Nutrition plays a vital role for normal growth and to maintain physical and mental
fitness throughout one’s life. Inadequate nutrition may lead to malnutrition, growth,
retardation, reduced work capacity and poor mental and social development (Awasthi
and kumar, 1999).

Nutrient deficiencies/ malnutrition i.e., deficiencies of vitamins, protein and other

important nutrients considered the most widespread form of malnutrition, with
women and children being particularly vulnerable. It is a major impediment to
socioeconomic development and contributes to a vicious cycle of underdevelopment,
to the detriment of the already underprivileged groups.

In growing children, the adverse effects of micro-nutrient deficiencies include poor

growth and development, mental and neuromotor performance, immuno competence,
physical working capacity, overall reproductive performance as well as increased
morbidity, mortality, and risk of maternal death (Viteri FE and Gonzalez H 2002).

In many low-income countries including Ethiopia, large proportions of the population

are nutritionally vulnerable (Tontisirin et al., 2003). Generally, infants and young

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children, pregnant and lactating women, and the elderly are regarded as nutritionally
vulnerable.

Vulnerable groups are population groups whose immune system is compromised or

depressed nutritionally, medically, or socially and require provision of extra,
nutritionally high-quality foods in addition to the general ration to rehabilitate or
prevent deterioration in their conditions; beside various problems being faced by the
vulnerable groups.

Malnutrition in Ethiopia derives directly from dependence on un diversified

livelihoods based on low-input, low-output rain-fed agriculture. Ethiopian farmers do
not produce enough food even in good rainfall years to meet consumption
requirements (Stephen Devereux).Overcoming micro-nutrient malnutrition is
therefore, a precondition for ensuring rapid and appropriate national development.

The search for novel high quality but cheap sources of protein and energy has
continued to be a major concern in many parts of the developing world (Arinathan et
al., 2009). Investigations on economically viable indigenous food ingredients as
alternative strategies to curb under nutrition and food insecurity are of utmost
importance to broaden the essential nutrient sources for human nutrition (Barba de la
Rosa et al., 2009)

A good number of global goals have been established and considerable investments
have been made by governments and aid agencies in programs designed to prevent
micro-nutrient malnutrition in Ethiopia and other developing countries.

Diet-based strategies through consumption of a broad variety of foods are the most
promising approach for a sustainable control of malnutrition and disease among the
vulnerable groups (Marchione, 2002); Vulnerable groups are population groups whose
immune system is compromised or depressed nutritionally, medically, or socially and
require provision of extra, nutritionally high-quality foods in addition to the general
ration to rehabilitate or prevent deterioration in their conditions.

Beside various problems being faced by the vulnerable groups, cancer and diabetes
have been increasing dramatically in the last two decades and the

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prevention/treatment of diabetes has received a paramount importance among the
health professionals and nutritionists.

The easiest and cheapest way of overcoming micro-nutrient malnutrition for most of
the vulnerable groups of the population is the blending of our locally available food
crops to produce high nutrient, inexpensive and popular food products with high
quantity/quality of micro-nutrients, to provide alternatives to the more expensive
imported foods.

The leaves, seeds, and flowers of MORINGA oleifera all have great nutritional and
therapeutic value. The seeds are eaten like peas or roasted like nuts; while the flowers
are eaten when cooked and taste like mushrooms. The leaves are outstanding as a
source of vitamins A, B group and (C when raw) and are among the best sources of
minerals.

They are also excellent sources of protein, but poor sources of carbohydrates and fats.
Thus, MORINGA leaves are one of the best plant foods available in nature (Price LL.
2000).

Therefore the aim of this project is, to study MORINGA leaf powder addition on the
nutrient composition of soybean and peanut flour formulations for possible use as
supplementary food.

1.3 Objectives

1.3.1 General objectives

The general objective of this study is to develop and characterize a fortified blended
food (FBF) from peanut, soybean and MORINGA leaf powder (MLP).

1.3.2 Specific objective

❖ Evaluate the effects of MORINGA powder .on the nutritional quality of FBFs.

❖ To evaluate proximate composition of the food product.

❖ To test the product for bio active compounds.

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❖ To evaluate the sensory acceptability of the product.

1.4 Scope of the study

The scope of this study is limited to the development and characterization of

MORINGA fortified blended food in powder form from the specified local raw
materials.

1.5 Significance of the study

This research work is generally expected to be beneficial in the following ways:

❖ Experience for the researchers.

❖ Serve as a reference material for other researchers doing work on similar topic.

❖ Development of MORINGA fortified food.

❖ Help alleviate malnutrition in the society.

❖ Indicate the effect of blending on the quality of food

❖ Development of a new product from locally available materials.

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 CHAPTER TWO

2 Literature review

2.1 Malnutrition

Malnutrition refers to deficiency of all essential nutrients. In the world each country is
affected by different forms of malnutrition, especially in developing countries due to
poverty and lack of sufficient knowledge about nutrition, malnutrition is the key cause
of morbidity and mortality of children in the world such as 149 million under five
years children are attacked by stunned, 45 million wasted, 38.9 million
overweight,45% death (WHO, 2021).There are two types of malnutrition, the first one
is protein-energy malnutrition which includes lack of protein, fat, and carbohydrate,
the second one is micro-nutrients such as deficiency of essential minerals and
vitamins. Malnutrition causes adverse effects on up to two years of age infants
because it is at this age children have high nutritional requirements for growth and
development (Blossner et al., 2005), high brain growth velocity, this age is a transition
period of family food trained so it must be fulfilled required quality and quantity of
nutrients. Under nutrition included underweight (low weight for age), wasting (low
weight for height), and stunting (low height for age). 2.2.1

2.2 Malnutrition in Ethiopia

Malnutrition is a common public health problem and the main cause of death of
children in developing countries. In Ethiopia there are different forms of malnutrition
such as low weight for height (Wasting), low height for age (Stunning), low weight
for age (underweight) and the second form is micro-nutrient malnutrition like vitamin
A deficiency, Iron deficiency, and Zinc deficiency which includes overweight and
obesity which is too heavy or excessive fat for height. In Ethiopia, the main causes of
malnutrition are determined food insecurity, poor guiding and child feeding practices,
high incidence of infectious diseases, and limited access to quality nutrition services.
Household capital, education, and family planning are also key drivers of children’s
nutrition.

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2.3 Production and usage of soybean, peanut and MORINGA

2.3.1 Peanut
Groundnut (Arachishypogaea L.), which is also known as peanut, earth nut, monkey
nut and goobers, is an annual legume. It is one of the world’s most important oil seed
crops (Dwivedi, S.L., Crouch et al 2003), ranking the 13th most important food crop
and 4th most important oil seed crop of the world (Surendranatha, E.C., Sudhakar, et
al 2011), being cultivated in more than 100 countries in six continents (Sharma, K.K.
and Mathur, B.P., 2006).

Cultivated groundnut originated from South America (Weiss, E.A., 2000.) Its
cultivation is mostly confined to the tropical, subtropical, and warm temperate (zones)
countries between 40o N and 40o S latitude. It is currently grown on 25.2 million
hectares worldwide with a total production of 35.9 million metric tons, with
developing countries in Asia (66%) and Africa (25%) as the major producers (FAO
2006). In 2009, China, India and the United States were the three largest producers of
groundnut (USDA-FAS 2010).

Groundnut is relatively new to Ethiopia. It was introduced from Eritrea to Hararghe in

the early 1920s by Italian explorers (Daniel, E., 2009). Major groundnut producing
areas in Ethiopia are Babile, Gursum, Beles, Didessa, Gambella and Pawe. Gamu
Gofa, Illubabor, Gojam, Wello and Wellega are identified as potential production
areas (Daniel, E., 2009). During the 2014, it was cultivated on 79943.03 ha of land
and 112088.7 tons of groundnuts were produced, with average yield of 1.402 tons per
ha (CSA 2014).

Peanut is the fourth most widely cultivated oil seed in the America continents, Africa,
and Asia. Peanut plants grow best in well-drained sandy soils and sunny warm
temperatures with moderate rainfall. In Ethiopia, the total planted area was 443000 ha
and a production of 103.7 M ton of grain as it has been reported in the 2014 cropping
season (FAOSTAT 2014). This report also showed that the productivity of peanut has
been 1604.1 kg ha 1. To increase this productivity, plant breeder has tried to release
new high yielder varieties of peanut in Ethiopia (Kebede and Bushra 2012). So far,
peanut varieties improving program in Ethiopian has obtained the peanut variety

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yielding up to 2.2-ton ha−1. When its productivity is comparing to the productivity
(3.25-ton ha−1) that has been reported in North Carolina is very low (Jones 2003).
Hence, improving the production and productivity of peanut in Ethiopia using
environmentally friendly technologies is essential.

2.3.2 Soybean

Soybean (Glycine max) is one of the most important food plants of the world and
seems to be growing in importance as industrial and multipurpose crop. In Ethiopia,
soybean is a multipurpose most nutritionally rich crop as its dry seed contains the
highest protein and oil content. Thus, production of soybean in Ethiopia is very
essential to overcome malnutrition and partially compensate the expensive source of
animal proteins and as a source of income for small holder farmers.

Production of this crop is indispensable in the country to enrich the staple cereal based
food with sufficient and high-quality protein (Mekonnen and Kaleb, 2014). Soybean
is a drought tolerant crop that requires warm climates and is suitable for low to
medium altitudes (Ogema et al., 1988; Urgessa, 2015). Since its introduction in
Ethiopia in the early 1950s soybean has become one of the most important lowland
grain legumes in the country that is highly adapted to diverse agro ecological
conditions including areas of marginal to the production of most of other crops.

Furthermore, soybean is the primary source of edible oil globally with the highest
gross output of vegetable oil among the cultivated crops with total cultivated area of
117.7 million ha and total production of 308.4 million tons (FAOSTAT, 2015). In
recent years, production and area cultivated under soybean in the country has
increased trend (One of the reasons for soybean production increase is policy
measures taken by the government.

For example, GTP II plan has given focus for soybean production as industrial crop
and its production is expected to increase from 0.72 million quintals in 2015 to 1.2
million quintals by the year 2020 to meet the market demand by creating a linkage
with the industry and export market (GTP II, 2015). Soybean is one of the legume
crops introduced to Benishangul-Gumuz region during the resettlement program in
1986. Predominantly, the crop is produced by smallholder and some commercial

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levels in the region with productivity potential of 16 to 17 qt/ha (BoARD, 2014). The
productivity level of the crop is nearest to the national average yield which is around
17.2 qt/ha. The crop grows widely in those zones of the region mainly for its
economic advantage in the local market and household consumption (AsARC, 2006).

2.3.3 MORINGA

M. stenopetala, often referred to as the African MORINGA tree, is a multipurpose

tree native to Ethiopia, northern Kenya and eastern Somalia, which has a wide range
of adaptation from the arid to humid climates (Jahn, 1991). MORINGA stenopetala is
locally called Shiferaw in Amharic or Haleko in Gamo and Wolyeta, and cabbage tree
in English (Abuye et al., 2003).

The botanical description of this tree: the height is 6–12 m with a diameter of 60 cm, a
smooth bark, strongly branched, sometimes with several trunks, and its wood is soft
(Abuye et al., 2003; Orwa et al., 2009). This plant is drought resistant like in southern
part of Ethiopia and often grows in well-drained soils at altitudes of 900–1200 m with
a mean annual rainfall ranging from 500–1400 mm (Orwa et al., 2009), however, the
cold temperature is the limiting factor for the cultivation of this species.

2.3.3.1 Nutritional composition

MORINGA leaves have been advocated as a source of highly digestible proteins with
considerable amino acid profile that contains the sulfur-containing amino acids,
methionine and cysteine (Booth and Wickens, 1988). It is also rich in the minerals
calcium and iron and the vitamins A, B, C and E (Booth and Wickens, 1988). The
leaves are also rich in β-carotene and are an exceptionally good source of fiber
(Nambiar et al., 2003). M. stenopetala leaves are rich in protein (28.2%) and contain
reasonable amounts of essential amino acids, vitamins, and minerals (Abuye et al.,
2003; Melesse, 2011).

M. oleifera leaves are nutritious and contain an average of 29% protein, 28 mg of

iron, 1.9 g of calcium, and 0.8 g of vitamin C (Wangcharoen and Gomolmanee,
2013). The leaf also has a wide range of beneficial polyphenolic compounds, which
include zeatin, quercetin, β-sitosterol, caffeolquinic acid, rutin, lutein, catechins,

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isothiocynates and kaempferol (Nambiar et al., 2003). The seed also has a high oil
(41.1%) and protein content (42.6%) (Seifu, 2012). This shows that it can be used as
a source of oil. In addition, the seeds of this plant are used for water treatment (Bichi
et al., 2012). On the other hand, MORINGA leaf contains various antinutritional
factors that may affect efficient utilization, absorption and digestion of nutrients, and
thereby decrease their bioavailability and nutritional value (Lestienne et al., 2007).

In addition, the bitterness and dark green color (Abuye et al., 2003) is a limitation to
the use of MORINGA leaf powder in food formulations. According to the findings of
(Sengev et al 2013), wheat bread with 5% of MORINGA leaves flour was
unacceptable though it has high nutritional values. Moreover, a wheat cookies
developed by mixing MORINGA leaves flour, had acceptability to the maximum of
10% MORINGA flour mixture (Nwakalor, 2014).

2.3.3.2 Bio active compounds

Bio active compounds are essential and non-essential compounds (e.g., vitamins or
polyphenols) that occur in nature and can be shown to have an effect on human health
(Biesalski et al. 2009). The majority are found in foods that predominantly originate
from the plant kingdom - in various fruits, vegetables and grains - with some others
from animal sources (Rein et al. 2013).

Bio active compounds vary widely in their chemical structures and functions; as such,
they are categorized accordingly (Kris-Etherton et al. 2002). Several examples of
plant derived bioactive compounds include polyphenolics, organosulfur compounds,
phytosterols, carotenoids and monoterpenes - all of which have subcategories
exhibiting a diverse range of chemical and biological properties (Kris-Etherton et al.
2002). They may also differ in attributes regarding sites of action, distribution in
nature, and concentrations in foods as well as the human body (Carbonell-Capella et
al. 2014)

Natural antioxidants protect against various diseases, which are induced by free
radicals. Antioxidants have various activities in addition to scavenging free radicals.
These include inactivating metal catalysts by chelation, reducing hydroperoxides into
stable hydroxyl derivatives and interacting synergistically with other reducing

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compounds (Frankel and Finley, 2008). The increased popularity of natural food
additives may prompt food manufacturers to replace synthetic antioxidants with
ingredients containing natural anti-oxidative compounds.

2.3.3.2.1Polyphenols

Polyphenols are polyhydroxyphenols, which are structural class of compounds that

are mostly composed of natural compounds, and sometimes synthesized or semi
synthetic in nature (Cattani et al. 2012).

Polyphenols are among the largest group of secondary metabolites found in plants.
Over 8000 structurally variants of polyphenols exist which are characterized by
aromatic rings with one or more hydroxyl groups (Han et al. 2007). These compounds
have the characteristics of multiple phenolic structural units in them, hence, the name
polyphenols. Polyphenols are mostly of plant origin and they are among the most
studies class of phytochemicals.

Some classes of polyphenols are

❖ Flavonoid

❖ Tanin

❖ Lignan

❖ Stilbenes

2.3.3.2.2Organosulfur Compounds

Organosulfur compounds (OSCs) are natural compounds that contains sulfur. They
are unique because of their unsavoury or foul smelling odours, with few exceptions.
This class of compound contains most of the Earth’s sweetest compounds derivatives
of sulfur, e.g. saccharin, a benzoic sulfimide artificial sweetener which is almost 400

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times as sweet as sucrose. OSCs are present naturally in plants and animals. They are
indispensable to life because they help in the prevention and treatment of several
life-threatening diseases such as cancer, diabetes, cardiovascular diseases,
neurodegenerative disorders, viral, bacterial and fungal infections. The natural world
has abundant of OSCs. Among the 20 general amino acids, two (methionine and
cysteine) are OSCs, while the antibiotics sulfa and penicillin medicines both have
sulfur (Block 1978)

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Bio active compounds and their sources

No Sources Bioactive
compounds

1 Fruits Apple, grape, Polyphenols,

cherry, peach, anthocyanins,
mangos, blueberry, carotenoids,
cranberry, flavonoids,
raspberry, bilberry, betalains
cactus pear (betacyanins,
betaxanthins)

2 Vegetables Carrot, tomato, Carotenoids,

onion, cauliflower, lycopene,
broccoli polyphenols,
glucosinolates,
vitamins

3 Grains Pigmented corn, Anthocyanins,

oat, barley, wheat, polyphenols,
amaranth, bean, flavonoids, soluble
rice fiber, l-lysine

4 leaginous by-products Defatted soybean Proteins,

paste, pumpkin, polyphenols,
and defatted carotenoids
sunflower pasta

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2.3.3.2.3Overview of Bio availability of nutrients and bio active
compounds

The ability of a bio active compound to exert its health-promoting effects upon a
living organism depends on its bio available dose instead of the administered dose
(Holst and Williamson 2008). The bio available dose is a more accurate representation
of what the body is capable of absorbing, distributing, metabolizing and excreting
(ADME) as opposed to what is administered or how much of a compound is present
in specific foods.

A prerequisite to assessing bio availability is bio accessibility, or the fraction of a food

constituent that, as a result of being released from the food matrix, becomes present in
the gut and may pass through the intestinal barrier (Saura-Calixto et al. 2007). A
compound must be released from the food matrix, whether by digestive enzymes or
colonic micro flora, and in the right form to be absorbed before it can become bio
accessible (D'Archivio et al. 2007). The caloric content and makeup of the food
matrix can affect the bio accessibility of digested compounds.

2.4 Fortified blended foods

Fortified-blended foods (FBFs) are porridge mixes composed of cereals and legumes
that have been milled and fortified with vitamins and minerals. FBFs are major food
aid products for young children, women, and other vulnerable groups in developing
countries. Historically, corn-soy blend (CSB) has been the most widely distributed
FBF in a majority of the food aid–receiving countries (Fleige LE et al 2010). The US
Agency for International Development (USAID) Food Aid Quality Review (FAQR)
recommended developing novel FBFs using cereals that are both culturally and
nutritionally acceptable in Africa. It also recommended sorghum as an alternative to
corn or wheat and suggested other legumes could be paired with it as alternatives to
soy (Webb P, Rogers B et al 2011). One logical legume to investigate is cowpea,
because Africa is the world’s leading producer of cowpea (95%) n addition to
sorghum (41%).

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FBFs, that are currently used, are partially pre-cooked foods. They are designed to be
cooked, fried or baked to complete their digestibility. WFP (2002) suggested that the
cooking time for FBFs should vary from 2 to 15 minutes depending on the kind of
preparation required. Vegetables, seasoning and other additives are used in order to
improve the palatability and to increase the nutritive value of the final product. (Rowe
et al. 2009) reported that African people added sugar and vegetable oil to their meal.
Cinnamon, herbs, or banana were often added to the Guatemalan recipes. Thin or
thick porridges are the most common dishes prepared from cereal-based products
(Rowe et al., 2008; Moussa et al, 2011). The difference between thick and thin
porridge is the concentration of the flour used in the preparation.

Advantages of FBF

❖ It is of low cost

❖ Readily provides amino acids

❖ The blended flour ingredient is readily available

Disadvantages

❖ The pre-mixing of the ingredients is a difficult procedure

❖ The product is bulky (i.e. it’s relatively of a high weight for the number of

calories provided compared to other products)

❖ It is of poor nutritional quality

❖ Requires an addition of oil which may not be readily available

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 CHAPTER THREE

3 Materials and method

3.1 Description of the study area

This experiment was carried out in wolkite university food engineering laboratory;
wolkite is the capital of gurage zone in SNNPR and the town is 170km south west of
Addis ababa.

3.2 Sample Collection and Transportation

Matured MORINGA leaves were bought from the local market of Sawla town in Gofa
zone. The leaves were fresh green colour. Leaves striped off the branches before
transporting them to the laboratory in wolkite town. The leaves were transported in a
carton box instead of plastic bags to allow sufficient aeration to prevent quality loss
loss. The remaining raw materials were provided by wolkite university laboratory.

3.3 Raw material preparation

MORINGA, Soybean and Peanut flours are prepared using a modification of the
method described by Solomon. The clean green MORINGA leaves, soybeans and
peanut will be washed separately in clean tap water. The MORINGA leaves and
peanut are dried in an oven for 15 minutes; while MORINGA leaves were dried in a
solar drier.

3.3.1 Soybean flour preparation

The soybean is soaked in tap water for 10 minutes to remove the hull and then dried
in an oven at 110˚C for 15 minutes; soybean is separately milled in a disc attrition
mill (ASIKO All, Addis, Nigeria) and sieved using a 500 µm sieve. and packed for
further use. (Soaking and roasting were intended to remove the beany flavour).

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Sorting and soaking

Drying

Sieving

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3.3.2 Peanut flour preparation
The peanut is also roasted in an oven at 70˚C for 15minutes and the seed coat
removed to get clean partially roasted peanuts. Peanut flour is obtained by pounding
the roasted peanut in a wooden mortar for size reduction and milling in a 2L mistral
grinder (model SAISHO, S-T4PN) into a smooth powder.

3.3.3 MORINGA powder preparation

MORINGA leaf powder will be prepared using a modification of the method

described by Gernah and Sengev . MORINGA leaves were washed in clean tap water
containing 5% Sodium chloride. They are then dried a solar drier, grounded into
powder and sieved with a fine sieve (500 µm). The powder is then dried to constant
weight in a hot air oven at 60˚C. All flours were packaged in black polyethylene bags
and stored in air tight plastic containers away from light.

3.3.3.1 Washing

The leaflets are washed in troughs using clean potable water to remove dirt. Washing
is done again in 1% saline solution for 3-5 minutes to remove microbes.

Finally washed again in clean water. Leaves are now ready for drying. Drain each
trough after each wash: fresh leaves must always be washed with fresh water.

3.3.3.2 Drying

The leaves are spread thinly on a mesh and dried in the dryer for about 4 hours
(Temperature range is 35°C–55°C on a very sunny day). The final product should be
very brittle. solar drying is recommended for both small and large scale processing,
particularly for those in rural communities where there is no electricity. Loading
density should not exceed 2 kg/m2 .

3.3.3.3 Milling

Dry leaves are milled using a stainless steel hammer mill. For personal or household
use, leaves can be pounded in a mortar, or milled with a kitchen blender. Small-scale

23 | Page
processors can use a burr mill or rent a commercial hammer mill for routine milling of
their products.

3.3.3.4 Sieving

Sieve the leaf powder if need be. When milled with a hammer mill, the fineness of the
product will depend on the size of the screen used in milling. If too coarse, sift using a
sifter with the desired screen size.

Recommended particle sizes are:

Coarse ( 1.0 mm – 1.5 mm)

Fine (0.5 mm – 1.0 mm)

Very fine (0.2 mm – 0.5 mm)

3.4 Functional properties of flours

3.4.1 Water Absorption Capacity (WAC)

Water absorption was measured according to the procedure of (Andersson et al).

(1979) with some modifications. About 2g sample was added to 1 ml distilled water
in a weighing centrifuge tube. Material was suspended in water by mixing with a thin
glass rod without meal adhering to the side of the centrifuge tube. After a holding
period of 30 min, 10 ml of distilled water were used to wash the meal adhering to the
stirring rod and centrifuge tubes. The suspension was then centrifuged at 3000 rpm for
25 min. The supernatant liquid was discarded and the tube was kept mouth down in
forced draft air oven at 50°C for 20 min, it was allowed to drain and dry, and then it
was kept in desiccators at (22±3°C) and subsequently weighted. Water absorption WA
expressed as the amount of water retained by 100 g of the sample. Water absorption
index (WAI) and water solubility index (WSI)

WAI (grams of gel recovered per gram of dry sample) and WSI (percent of dry
sample in water layer) were determined according to the methods of Andersson and

24 | Page
Hedlund (1991). Two grams of the sample were suspended in 30 ml of water in a 50
ml centrifuge tube at 30°C., then stirred intermittently for 30 min, and centrifuged at
3000 rpm for 10 minutes. The supernatant liquid was carefully poured into a tared
evaporating dish. The remaining gel was weighed and the WAI was determined. The
amount of dried solids recovered by evaporating the supernatant from the water
absorption test was expressed as percentage of dry solids.

WSI and WAI were calculated according to the following equations:

𝑠𝑜𝑙𝑖𝑑𝑠 𝑤𝑒𝑖𝑔ℎ𝑡
WA = 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒
×100

𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑔𝑒𝑙
WAI= 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒
×100

3.5 Proximate Analysis

The physicochemical analysis for the final product will be done using the AOAC
method. The determination method of the moisture, carbohydrate, protein, fat, fiber,
and ash is shown below.

3.5.1 Moisture Content determination

Moisture content will be determined by the AOAC method (Association of Official

Analytical Chemists) (AOAC, 2000); the Official Method 925.10, by drying the
samples in an oven until a constant weight is obtained. The crucible with its content
will put into a drying oven at 105°C for 5hr and place in desiccators to cool. The
weight of the crucible (W1) will be determined. 5 grams of the samples will
accurately weigh into a previously cleaned, dried and weighed glass crucible. Samples
will weigh in dry crucible (W2) will dire at 105 º C until constant mass will attain and
after cooling in desiccators to room temperature it will again weigh (W3). The
samples will then cool in desiccators and weigh. The process will be repeated until a
constant weight is obtained.

25 | Page
The loss in weight will be expressed as a percentage of the initial weight of sample
gave the percent moisture content. The moisture content will be determined using
equation(Tolesa, 2014)

𝑤2−𝑤3
Moisture content (%) = 𝑤1−𝑤2
×100

3.5.2 Total Ash determination

Total Ash content was determined by the method specified by Association of Official
Analytical Chemists‟ (AOAC, 2000), using the Official Method 923.03. Clean
porcelain crucible, dried at 120⁰C in an oven will ignite at about 550⁰C in a muffle
furnace for 3 hours will be cooled in desiccators and weigh (M1). Then 2.0 g samples
will be weighed into previously dry and weigh (M2) porcelain crucible. These
samples will dry at 120⁰C for 1 hour and carbonized by oven until the contents turn
black. The crucible with the contents will place in a Muffle furnace (Gallenkamp,
Model: FSL 340-0100, U.K) set at 550⁰C for 1 hour to ignite until ashing is
completed. After this period the crucible with its content is removed and cooled in the
desiccators. The crucible with the residue is weighed (M3). The weights of the ash are
expressed as a percentage of the initial weight of the samples.

𝑤2−𝑤3
Total Ash (%) = 𝑤2−𝑤1
×100

Where: (M2 – M1) is sample mass in g on dry base and (M3 – M1) mass of ash in g

3.5.3 Determination of Crude Protein

Crude protein was determined by the method of the Association of Official Analytical
Chemists‟ (AOAC, 2000) using the Official Method 920.87. The determination of
crude protein will pass through three steps these are digestion, distillation and
titration.

Two grams (2.0g) of the samples is weighed into a digestion flask and 0.5g of
selenium catalyst is added. 25ml of concentrated H2SO4 is then added and the flask
shaken to mix the contents. The flask is then placed on a digestion burner for 8h and
heat to (370⁰C) allowing digestion until the solution turns green and clear. The sample
solutions are then transferred into a 100ml volumetric flask and made up to the mark

26 | Page
with distilled water. Then 25ml of 2% boric acid is added using a pipette into a 250ml
conical flask and 2 drops of mixed indicator (20ml of bromocresol green and 4ml of
methyl red) solution will add. Into the decomposition chamber of the distillation
apparatus will add 15ml of 40% NaOH solution. 10ml of the digested sample solution
will then introduce into a flask. The condenser tip of the distillation apparatus will
then dip into the boric acid contained in the conical flask. The ammonia in the sample
solution will then distill into the boric acid until it completely changed to bluish
green. Finally, the distillate will titrate with standardized 0.1N Sulphuric acid to a
reddish color. The percent total nitrogen and crude protein is calculated using the
following equation.

(𝑉2×𝑉1)×𝑁×14.007
Nitrogen (%)= 𝑊°
×100

Where:

❖ V2 = Volume in ml of standard sulfuric acid solution used in the titration for the

test material. 23

❖ V1 = Volume in ml of standard sulfuric acid solution used in the titration for the

blank determination.

❖ N = Normality of standard sulfuric acid (0.1N).

❖ Wₒ = Sample weight on dry matter basis and 14.007 is the molecular weight of

nitrogen.

N. B: The % of nitrogen is converted to % of protein by using appropriate

conversion factor (Tolesa, 2014)

(Crude protein content percent per weight = total nitrogen * 6.25 for composite flour
and total nitrogen 5.71 for wheat flour and banana flour).

27 | Page
3.5.4 Crude Fat Determination

Crude fat was determined based on the Sohxlet extraction method of AOAC (2000)
using official method 920.39. A 250 ml quick fit round bottom flask will wash and
dry in an oven (Gallenkamp, model OV 880, England) at 105⁰C for 25 minutes and
allow to cool to room temperature before it is weighed. A clean and dry muslin
thimble containing about 5 g of dry sample and cover with fat free cotton at the
bottom and top is placed in the extraction chamber. 2.0g of the samples will be
weighed into the thimble. This is then inserted into the extraction column with the
condenser connected. 200ml of the extracting solvent (petroleum ether, boiling point
40-60⁰C) will pour into the round bottom flask and fit into the extraction unit. The
flask will then heat with the aid of electro-thermal heater at 60⁰C for 8 hrs. Losses of
solvent due to heating will check with the aid of the condenser so that it cool and
reflux the evaporated solvent. After extraction, the thimble will remove and the
solvent salvaged by distillation. The flask containing the fat and residual solvent will
place on a water bath to evaporate the solvent followed by a further drying in an oven
(Gallenkamp, model: OV 880, England, 1974) at 1050C for 30 minutes to completely
evaporate the solvent. It will then cool in desiccators and weigh. The flask containing
the extracted fat will dry on a steam bath at 98⁰C to a constant mass. The fat obtained
will express as a percentage of the initial weight of the sample using the following
formula.

𝑊2−𝑊1
Crude fat, % by weight= 𝑊
×100

Where:

❖ W1 = weight of the extraction flask (g),

❖ W2 = weight of the extraction flask plus the dried crude fat (g), and

❖ W = weight of samples (g)

28 | Page
3.5.5 Crude Fiber determination

Crude fiber is determined by the method of the Association of Official Analytical

Chemists‟ (AOAC, 2000) using the official method 962.09. About 3.0g defatted
samples (from crude fat determination above) are transferred into 750 ml Erlenmeyer
flasks and 200ml of boiling 1.25% H2SO4 will add and the flask will immediately set
on a hot plate electric oven at 130⁰C and condenser connected to it. The content will
brought to boil within 1 minute and the sample will digest for 30 minutes. At the end
of the 30 minutes, the flask is removed and the content is filtered through a linen cloth
in a funnel and subsequently will wash with boiling water until the washings is no
longer acidic.

The samples will wash back into the flask with 200ml boiling 1.25% NaOH solution.
The condenser will again connect to the flask and the content of the flask will boil for
30 minutes. It will then filter through the linen cloth and thoroughly wash with boiling
water until the washings will no longer alkaline. The residue will transfer to a clean
crucible with a spatula and the remaining particles wash off with 15ml ethanol into
the crucible. The crucible with its content will then dry in an oven (Gallenkamp,
Model: OV 880, England, 1974) at 105⁰C overnight and cool in a desiccator and
weighed (M1). The crucibles with its content will then ignite in a furnace (Muffle
furnace size 2, England) at 550⁰C for 2h, cool and re-weigh (M2). The loss in weight
gave the crude fiber content and will express as a percentage of the initial weight of
the sample using the formula. The total crude fiber will expressed in percentage as

𝑀1−𝑀2×100
𝑡𝑜𝑡𝑎𝑙 𝑐𝑟𝑢𝑢𝑑𝑒 𝑓𝑖𝑏𝑒𝑟 (%) = 𝑀3

3.5.6 Determination of Total Carbohydrate

Total percentage carbohydrate was determined by the difference method as reported

by Osborne and Voogt (1978). This method involves adding the total values of crude
protein, crude fat, crude fiber, moisture and ash constituents of the sample and
subtracting it from 100. The value obtained is the percentage carbohydrate constituent
of the sample. Total carbohydrate content of the samples including crude fiber will
determined by subtraction of the above test parameters from 100%. Carbohydrate
content will determine by difference.

29 | Page
Total %C= 100 − (%𝑀 + %𝑃 + %𝐹 + %𝐹𝐵 + %𝐴)

Where:

❖ C=Carbohydrate content,

❖ M=Moisture content,

❖ P=Protein content,

❖ F=Fat content,

❖ Fb=Fiber content and

❖ A=Ash content.

3.5.7 Calorific Value

Energy value (calorific value) was quantified using an indirect calculation method.
The three groups of nutrients, which provide the body with energy, are carbohydrates,
fats and proteins (James, 1995). One gram of carbohydrate (C) was assumed to give
4Kcal energy; one gram of fat (F) 9Kcal energy and one gram of protein (P) 4Kcal.
Therefore, determination of calorific value (Kcal/100g) of baked products will
determine according to James (1995).

Where:

❖ P = Protein content (%).

❖ F = Fat content (%).

30 | Page
❖ C = Available total carbohydrate (%)

3.6 Test for bioactive compounds

3.6.1 Condensed tannins

Condensed tannins were determined according to a SOP. The method is based on

Price et al. (1978) using vanillin reagent. For this method, methanol extracts were
made of leaf and leaf-derived samples. The extracts were mixed using an ultra-turrax
(45 seconds at 10000 rpm) and centrifuged (15 minutes, 4000 rpm, 4 °C). Both the
pellet and the supernatans were collected. The analysis was performed on the
collected supernatans.

3.6.2 Phytic acid content

The method for this analysis is described by Reichwald & Hatzack (2008) and is
based on determining phytic acid through indirect spectrophotometry, as the
absorbance by light of non-complexed iron to phytate is measured. A standard curve
with concentrations ranging between 0 mg/ml and 140 mg/ml was constructed using
phytic acid dodecasodium salt. The absorbance was measured at 540 nm.

3.6.3 Total phenolic compounds

Total phenolics were determined using the Folin-Ciocalteu method (Waterhouse 2002)
with modifications. Gallic acid (Sigma-Aldrich Co., St. Louis, Mo., U.S.A.) standards
were prepared with 0.0, 50.0, 100.0, 150.0, 250.0, 500.0 mg/L. Approximately 0.1 mL
of standard solution and hydrophilic extract samples were pipetted into test tubes, to
which 7.9 mL of deionized water and 0.5 mL Folin reagent (Sigma-Aldrich Co.) were
added to each of the standard and sample solutions. After 1 min, 1.5 mL of sodium
carbonate solution was added followed by vortexing. The sodium carbonate solution
was prepared with 100 g anhydrous sodium carbonate in 400 mL water, which was
allowed to sit for 24 h, filtered and brought to a final volume of 1 liter. Standards and
samples remained at room temperature for 2 h followed by absorbance measurement
at 765 nm using a SAFIRE2 microplate reader equipped with version 6.1 Magellan
reader software (Tecan US, Raleigh, N.C., U.S.A.). Total phenolics were calculated as
milligrams of gallic acid equivalents per gram (mg GAE/g)

31 | Page
3.7 Sensory evaluation of FBF as a drink

Sensory analysis attribute of the food product was done in wolkite University,
Institute of Technology, 18 people were selected as panelists to do the analysis The
panelists were given orientation on overall sensory evaluation procedures. The
powder was prepared into a shake by directly mixing with powdered milk, sugar and
water then served to the panelists with small coffee cups. This evaluation included
color, Aroma, sweetness, appearance taste, bitterness, and overall acceptability using
seven hedonic scale;

(7) like extremely,(6) like very much, (5)like slightly, (4)neither like nor dislike,
(3)dislike slightly, (2)dislike very much,(1)dislike extremely.

The Panelists were provide with clean bottled water to rinse their mouth with after
evaluating each sample.

3.8 Production of experimental FBF

3.8.1 Formulation of experimental food blends

When deciding the blend ratio of each raw material proximate composition of the
product was the main criteria. Six blends were formulated, out of one, (50% peanut,
50% soybean) which had no moringa powder served as the control.

The five experimental blends were prepared according to the percentages of

constituent ingredients given in the following table and each blend is named
according to its distinguished constituent ingredient as follows:

Percentage of ingredients used to develop the snack products

Ingredient blending ratio

Sample Peanut (%) Soybean (%) MORINGA (%)

code

G1P1 50 50 0

32 | Page
G1P2 45 50 5

G1P3 50 45 10

G1P4 40 45 15

G1P5 45 35 20

G1P6 35 40 25

33 | Page
Production process flow chart

Moringa leaves

34 | Page
 Soybean

Peanut

Washing

Sorting

Sorting

Drying in solar dryer

Soaking

Roasting

Grinding

De-husking

De-husking

Blanching

Grinding

Drying

Sieving

Grinding

Mixing

Sieving

Blended product

3.9 Experimental design and statistical analysis

Experimental analysis of results was given as mean and standard deviation (SD) using
one-way analyses of variance (ANOVA) by using SPSS version 20 Duncan‟s multiple
range test software application. Except for sensory attribute analysis, some analysis

35 | Page
was measured in triplicate of each sample and some were in duplicate, differences
considered to be significant at P<0.05 (95% confidence level).

In this study optimization of proximate composition analysis were by using Minitab

17 software design the effect of a single variable (Factor) blending ratio (R1-R6) of
the product compared with the control blend to clearly understand the effect of
moringa fortification on proximate composition.

36 | Page
 Chapter four

4 Results and discussion

4.1 Proximate composition of the formulated FBF

This study focused on supplementary food formulated from a blend of soybean,

peanut and moringa leaf powder (MLP). by preparing five blends each with differing
amount of MLP to determine the effect of moringa supplementation on nutritional
quality and identify the blend with best nutritional value.

Proximate composition of formulated products was dependent on the nutrient

composition of raw materials used. The proximate compositions of formulated
products were compared to a control product which had no moringa in it.

37 | Page
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