EFFECT OF Moringa oleifera LEAVES ON THE ANTIOXIDANT AND NUTRITIONAL COMPOSITION OF

COMPOSITE BREAD
1Aishat M. Osagie, 2Nicholas O. Orobator and 3Marshall A. Azeke
Xenobiotic and Nutraceutical Research Group, Department of Biochemistry, Faculty of life sciences,
Ambrose Alli University, Ekpoma, Nigeria.

*Corresponding Author: aisha.osagie@aauekpoma.edu.ng

ABSTRACT
Background: Moringa oleifera commonly referred to as miracle plant is rich in healthy antioxidants and bioactive
plant compounds. The nutritional and health benefits of Moringa oleifera have been documented. However, fortifying
bread with Moringa leaves is a novel area that is yet to be fully exploited. Consequently, this study was aimed at
developing sustainable, non-perishable and healthy food thereby improving food safety and quality by evaluating the
antioxidant activities and nutritional components of composite bread with moringa oleifera leaf powder (MOLP).
Methods: Fresh moringa oleifera leaves were obtained, dried and blended into powder. Bread samples were prepared
by supplementing wheat flour with MOLP at the levels of 3.0, 5.0, 7.0, 10.0 and 15.0 % and their in vitro antioxidant
activities and nutritional components were determined using standard protocols.
Results: fortifying bread with MOLP resulted in a steady increase in the phytochemical constituents and antioxidant
activities as the proportion of MOLP to wheat flour increased. The results showed that, total phenols content, total
flavonoids and total alkaloids were significantly (p < 0.05) high in the composite bread made with 10% MOLP. While
total tannins, total saponins and FRAP activities were significantly (p < 0.05) high in the composite bread made from
7% MOLP. The result also showed that composite bread made with 10% MOLP was highest in reducing power
potential and DPPH radical scavenging ability. The result of the nutritional analysis showed that the crude fiber, crude
protein content and crude fat increased significantly (p < 0.05) as the MOLP to wheat flour increased. The
Carbohydrate content of the bread samples decreased significantly (p < 0.05) with increase in the MOLP of the bread
while the moisture content of the bread samples varied significantly (p < 0.05).
Conclusion: The results of this study indicate that supplementation of bread with Moringa oleifera leaf powder
improves the antioxidant and nutritional composition of the bread and can therefore be incorporated as functional food
for the management of diseases and improvement of health status.

Key words: Moringa leaves, composite bread, antioxidant activities, Nutritional composition.

INTRODUCTION
Food safety and nutrition security is a major global challenge which relies on the adequate supply of safe, affordable
and nutritious fresh and processed foods to all people. The challenge of supplying healthy diets will in part be met
through increase in food production. However, reducing food losses throughout the supply chain from production to
consumption and sustainable enhancements in preservation, nutrient content, safety and shelf life of foods enabled by
food processing will also be essential. Medicinal plants present abundant sources of natural antioxidants and find
enormous application in human nutrition, not only as flavoring spices but also as natural remedy. Herbal fortification
of bread and baked foods is a new trend to improve its nutritional value. Herbs are rich in minerals, vitamins,
flavouring agents and natural antioxidants. This supplementation of herbs adds a spicy flavour, greatly improved taste
and sensory properties, and enhanced the level of natural antioxidants. Consequently, this study is aimed at developing
sustainable, non-perishable and healthy food thereby improving food safety and quality and reducing post-harvest
losses.

According to World Data Atlas, vegetable consumption in Nigeria rose from 120g to 165g per person per day from
1992 to 2007. This trend has continued due to increased awareness of the health benefits of fruits and vegetables.
Despite this trend, a recent report by the Agricultural Fresh Produce Growers and Exporters Association of Nigeria
(AFPGEAN) indicated that between 55% and 72% of fruits and vegetables grown in Nigeria perish before they can be
consumed. This is mainly due to the absence of appropriate post-harvest processing and storage facilities. Herbal
medicines are the mainstay of about 75–80% of the world population, mainly in developing countries, for primary
health care because of better cultural acceptability regarding compatibility with the human body and less side effects
(Kingsley and Marshall, 2014). Herbal fortification of baked foods is a new trend that will not only improve the
cultivation and utilization of vegetables and herbs but also improve the nutritional value and health benefits of baked
products. Baked foods such as bread, biscuits, buns, doughnuts and sausage rolls are basic staple in many homes in
Nigeria. In fact, they are more staple than rice, cassava products or any of the popular staples perceived as an exclusive
preserve of Nigerians culinary culture. All genders, all ages, all tribes, both the rich and the poor consume these
products, and it comes in various sizes, shapes, compositions and price tags to meet the needs of different categories of
consumers. It is a grab and go staple that goes with almost anything, that is why there is virtually no household in
Nigeria where these baked foods are not consumed.
Herbs are rich in minerals, vitamins, flavouring agents and natural antioxidants. Roots, stems, leaves or seeds of herbal
plants have long been used in cooking and in naturopathy all over the world. In a recent communication, the
fortification of white bread with coriander leaf powder was reported (Das et al., 2012). This supplementation imparted
a spicy flavour, greatly improved taste and sensory properties, and enhanced the level of natural antioxidants. Fennel
seeds have also been used for the fortification of bread which showed high moisture content in the crumbs, rich
antioxidant content and good consumer acceptability (Das et al., 2013).

Moringa oleifera (MO) (Moringaceae) is a fast growing tree found in India, the Sudan, sub-Saharan Africa, the Pacific
and Caribbean Islands and South America (Araujo et al., 2013; Warhurst, et al., 1997) and can be used as a food crop, a
source of vegetable oil and as a coagulant and adsorbent in water treatment (Warhurst, et al., 1997; Baptista et al.,
2012). All over the world, the nutritional value of the plant is well known and it is also used for the treatment of
different and unrelated diseases. Different parts and preparations of MO have been reported to be used in traditional
medicine and in the treatment of various conditions (Bakre et al., 2013).

The plants are used to treat lots of health problems like nervous disorders (such as muscle spasmodic, epilepsy,
headache, hysteria), also are used for anemia, skin infections, blackheads, anxiety, bronchitis, catarrh, chest congestion,
asthma, blood impurities, cholera, glandular, swelling, headaches, conjunctivitis, cough, diarrhea, eye and ear
infections, fever, abnormal blood pressure, pain in joints, pimples, psoriasis, respiratory disorders, scurvy, semen
deficiency, sore throat, sprain, tuberculosis, for intestinal worms, lactation, hypertension and diabetes(Horwat and
Benin 2011; Mishra, et al., 2012). The MO plant contains various amino acids, fatty acids, vitamins and nutrients
glucosinolates and phenolics (flavonoids, anthocyanins, proanthocyanides and cinnamates) (Akhtar et al., 2007). The
leaves of MO are considered to be a rich source of vitamins and minerals and exhibits strong antioxidant activity, often
attributed to the plants’ vitamins and phenolic compounds such asquercetin and kaempferol (Coppin et al., 2013). MO
organs were also used as nutritional supplement (Mendieta-Araica et al., 2011; Popoola and bembe 2013).
Wheat bread being a basic staple in many homes in Nigeria, increasing its nutritional and biological value cannot be
overemphasized. This study therefore uses the medicinal vegetable MO which is a rich source of bioactive compounds,
especially essential oils, terpenes, phenolic acids and flavonoids with proven antioxidant activity to fortify bread.

METHODS

Sample Collection

The Moringa oleifera leaves were collected from Eboakhula area of Ekpoma, Esan-west local government area, Edo
state, Nigeria. Identification was done by a botanist in the Department of Botany, Ambrose Alli University, Ekpoma,
Nigeria.
Preparation of composite flour
To obtain the flour blends for bread preparation, 100 g of wheat flour was mixed with 3.0, 5.0, 7.0, 10.0 and 15.0 % of
MO leaf powder.

Blend Formulation
The method described by Igbabul et al. (2019) was adopted in this study. Six different samples of blends were
produced and coded as M0, M3.0, M5.0, M7.0, M10.0 and M15.0. Sample M0 served as the control and contained
100% wheat. Samples M0, M3.0, M5.0, M7.0, M10.0 and M15.0 consisted of wheat/Moringa oleifera leaf flours and
the other ingredients for bread production.
Table 1: Blend formulations.

Samples

Ingredients M0 M3.0 M5.0 M7.0 M10.0 M15.0

Wheat flour (g) 100 97.0 95.0 93.0 90.0 85.0
Moringa oleifera leave flour (g) 0 3.0 5.0 7.0 10.0 15.0
Salt (g) 1.5 1.5 1.5 1.5 1.5 1.5
Yeast (g) 2.0 2.0 2.0 2.0 2.0 2.0
Sugar (g) 2.0 2.0 2.0 2.0 2.0 2.0
Deionized water (ml) 50.0 50.0 50.0 50.0 50.0 50.0

Baking Process / Bread Production

The six blends of composite flour were baked into bread using the straight dough method (Chauhan et al., 1992). All

dry ingredients were weighed and mixed using machine food processor (Kenwood KM 201, England) for about 10

minutes at low speed to obtain dough. The produced dough was covered with food wrapper to prevent excessive

moisture loss and left to rise at room temperature for 60 minutes. Then, the dough was punched and kneads again to

release the air inside The kneaded dough was then transferred into baking pans greased with plasticized fat and covered

with greased bread wrapper. The dough was allowed to ferment for 90 minutes at room temperature in the baking pans.

The fermented dough was then allowed to undergo proofing at 400C for 90 minutes and then baked at 250 0C for 30

minutes. The breads were cooled to room temperature and used for analysis.
Sample Extractions

Bread samples were sliced (3 cm width and 1 cm thickness) and air-dried for 24 hours. The dried material was blend to
obtain powdered bread samples. One gram each of the powdered samples was weighed and extracted in 100 ml of 80%
aqueous methanol for 24 h at 25 °C on an orbital shaker. The extract was further filtered using Whatman filter paper
(No. 1) and the filtrate obtained was centrifuged at 3500 rpm for 15 min. Thereafter, the supernatant will be collected
and was used for further studies.

Determination of Total Phenol Content

Total phenol content was determined according to the Folin and Ciocalteau’s method (1927). Concentrations (0.2 - 1

mg/mL) of gallic acid were prepared in methanol. Then, 0.5 mL of the sample (1 mg/mL) was mixed with 2.5 mL of a

ten-fold diluted Folin- Ciocalteau reagent and 2 mL of 7.5% sodium carbonate. The mixture was allowed to stand for

30 min at room temperature then absorbance read at 760 nm. All determinations were performed in triplicates with

gallic acid utilized as the reference control (Folin and Ciocalteau, 1927). Total phenol content of the extracts were then

extrapolated from the standard curve.

Determination of Total Flavonoid Content

The total flavonoid content was determnined using the method of Miliauskas et al., (2004). 2 ml of 2% AlCl3 in ethanol was

mixed with 2 ml of varying concentration of the extracts (0.1 – 1.0 mg/ml) in methanol. The absorbance was measured at 420 nm

after one hour incubation at room temperature. Similar concentrations of quercetin, the positive control were measured. The total

flavonoid content was calculated as mg quercetin equivalent/g of extract. Total flavonoids content of the extracts were then

extrapolated from the standard curve.

Determination of Total Tannins

Tannins content was dertermined by Folin-Denis method (Polshettiwar et al., 2007). The method is based on the

measurement of a blue colour formed by the reduction of phosphotungsto-molybdic acid by tannin-like compounds in

alkaline medium. One milliliter of extract (1mg/mL) and standard solution of Tannic acid (10-150 µg/mL) was made

up to 7.5mL with distilled water. Then 0.5 mL Folin-Denis reagent and 1mL of 7.5 % Na2CO3 solution were added.

The volume was made up to 10 mL with distilled water and absorbance was measured at 700nm. Total tannins content

of the extracts were then extrapolated from the standard curve and expressed as mg of Tannic acid equivalent /g of

extract.
DETERMINATION OF TOTAL ALKALOIDS CONTENT

Total alkaloid level of the extracts was determined following the method reported by Singh et al. (2004). The extract (1

mL) was mixed with 1 mL of FeCl3 solution (0.025 M FeCl3 in 0.5 M HCL) and 1 mL of 0.05 M of 1, 10-

phenanthroline in ethanol. The reaction mixture was incubated in hot water bath at 70 ± 2oC for 30 minutes. The

absorbance of red coloured complex formed was measured at 510 nm against reagent blank. Alkaloids content of

sample was extrapolated from the standard curve and expressed as quinine equivalent in mg/g of sample dry weight.

DETERMINATION OF SAPONINS CONTENT

Estimation of total saponins content was determined by (Makkar et al., 2007) based on vanillin-sulphuric acid

colorimetric reaction with some modification. About 50µl of bread extract was added with 250µl of distilled water. To

this, about 250µl of vanillin reagent, 800mg of vanillin in (10mL of 99.5% ethanol) was added and it was mixed well.

This solution was kept in a water bath at 60°C for 10min, it was cooled in ice cold water and the absorbance was read

at 544nm. Quinine was used as standard. The values were obtained from a standard curve and expressed as quinine

equivalent in mg/g of sample dry weight.

Determination of Reducing Power Potential

The reducing power was determined according the method described by Lai et al. (2001). One milliliter of different
concentrations of extracts (0.1-1.0mg/mL)in water was mixed with 2.5mL of 0.2M phosphate buffer, pH 6.6 and 2.5mL
of 1& potassium ferricyanide. The mixture was incubated at 50ºC for 20mins. Thereafter, 2.5mL of trichloroacetic acid
(10%) was added to the mixture to stop the reaction. Then 2.5mL of distilled water and 0.5mL of 0.1% FeCl 3 were
added and the absorbance measured at 700nm. Higher absorbance values indicated higher reducing power. Vitamin C
served as a positive control.

Ferric Reducing Antioxidant Power (FRAP)

The FRAP assay was carried out using a modified method of Benzie and Strain (1996).

To 1.5 mL of freshly prepared FRAP solution (25 mL of 300 mM acetate buffer pH 3.6, 2.5 mL of 10mM 2,4,6-

tripyridyls- triazine (TPTZ) in 40mM HCl, and 2.5 mL of 20 mM ferric chloride (FeCl3・6H2O)solution) was added to

o
1mL of the extracts at concentrations of 100 - 600µM. The reaction mixtures were incubated at 37 C for 30 min and the

increase in absorbance at 593nm was measured. FeSO4 was used for the calibration curve and ascorbic acid served as

the positive control. FRAP values (expressed as mg Fe (II)/g of the extract) for the extracts were then extrapolated from

the standard curve.

Determination of 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) Radical Scavenging Activity

DPPH is a stable free radial with red color (absorbed at 517nm). If free radicals have been scavenged, DPPH will

change its colour to yellow. This assay uses this character to show free radical scavenging activity. Radical scavenging

activity was done by a slightly modified method of Brand-Williams et al., (1995).

The following concentrations of extract were prepared: 0.002, 0.005, 0.01, 0.025, 0.05, 0.075, 0.1, 0.15, and 1mg/mL.

Ascorbic acid was used as standard, and the same concentrations were prepared as the test solution. All the solutions

were prepared with methanol. 2mL each of the prepared concentrations were placed into test tubes, and 0.5mL of\

1mM DPPH solution in methanol was added. The experiments were carried out in triplicates. The test tubes were

incubated for 15 minutes at room temperature, and the absorbance read at 517 nm. A blank solution containing the

same amount of methanol and DPPH was prepared and the absorbance read. Lower absorbance of the reaction mixture

indicates higher free radical scavenging activity.

The radical scavenging activity was calculated using the following formula:

DPPH radical scavenging activity (%) = [(A0−A1)/ (A0)] × 100

Where A0 was the absorbance of DPPH radical + methanol; A1 was the absorbance of DPPH radical + sample extract

or standard. The 50% inhibitory concentration value (IC50) is indicated as the effective concentration of the sample that

is required to scavenge 50% of the DPPH free radical.

3.2.5 Determination of Proximate Composition

The moisture content, ash, crude fibre, crude protein and fat content of the bread produced from wheat and moringa

oleifera leaf powder were determined using the method of AOAC (2012). Total carbohydrate was calculated by

difference.

RESULTS AND DISCUSSION

The quantitative phytochemical constituents of bread made with Moringa oleifera leaf powder (MOLP) are shown in

table 1. Phenolics and flavonoids are natural compounds found in many cereal grains. A review published in 2012

found growing consensus for the hypothesis that the specific intake of food and drink containing relatively high

concentrations of flavonoids may play a meaningful role in reducing the risk of cardiovascular disease (CVD).

Phenolics have many health benefits like antioxidantive, antiinflammatory, antimutagenic and anticarcinogenic

properties. It also maintains flavor, taste, color and prevention of their oxidation deterioration. The results for the total

phenol content of composite bread showed a significant (p < 0.05) steady increase in value as the concentration of

moringa leaves to flour increased. As reported by Nouman, et al. (2016) analysis of hydro-methanolic extracts of

Moringa, leaves revealed a wide range of phenolic compounds. Many authors also reported that the leaves of M.
oleifera fresh or dried are known to be excellent source of antioxidants and they have significantly higher antioxidant

content comparing to fruits such as strawberries known for high antioxidant contents (Moyo et al., 2012; Vongsak, et

al., 2013; Gopalakrishnan, et al., 2016). The addition of MOLP greatly enhanced the phenol content of bread from

0.5049 ± 0.008 mg GAE) / g extract for control (100% wheat flour) to 1.2375 ± 0.037 mg GAE / g extract of 10%

MOLP. This result is in agreement with that of Bourekoua et al. (2018). The addition of MOLP significantly enriched

the total phenol content of gluten-free products.

The results for the total flavonoid content of composite bread also revealed a steady increase as the concentration of

moringa leaves to flour increased. The flavonoids content significantly (p < 0.05) increased from 1.553 ± 0.004 mg

QE/g extract of the control (100% wheat flour bread) to 9.012 ± 0.247 mg quercetin equivalent (QE) / g extract of 10%

MOLP bread. A decrease was however observed in 15% MOLP bread.

The results for the total tannins content further showed the same trend of significant increase in value as the

concentration of MOLP to flour increased. However, the bread made with 7% (0.5579 ± 0.015 mg TAE / g Extract)

MOLP was highest in total tannins content when compared to the control (100% wheat flour bread; 0.2946 ± 0.026 mg

TAE / g Extract). The presence in moderate amount of tannins is important for its pharmaceuticals and therapeutic

significance.
The results for the total alkaloids content also showed that bread made with 10% MOLP recorded a significantly (p <

0.05) high value (13.48 ± 0.06 mg QE/ g Extract) when compared to the control (5.39 ± 0.189 mg QE / g Extract).

Saponins are a class of chemical compound found in many plant species especially in legume plants (Song and Hu,

2009). It poses a wide range of bioactive activities. The results for the total saponins followed the same trend as total

tannins. The results showed that bread made with 7% MOLP was highest in saponins content (4.523 ± 0.034 mg QE/ g

Extract) when compared to the control (100% wheat flour bread; 2.272 ± 0.032 mg QE/ g Extract).

In general, bread supplemented with MOLP recorded enhanced phytochemical constituents when compared to bread

made with 100% flour. These results are in agreement with that reported by Supriya et al. (2015).
Table 2: Total Phenols, Total Flavonoids, Total Tannins, Total Alkaloids and Total Saponins content of
composite bread made from MOLP.

Total Phenol Flavonoids Total Tannins TotalAlkaloids Total Saponins

Sample (mg GAE / g Extract) (mg QE / g Extract) (mg TAE/ g Extract) (mg QE/ g Extract) (mg QE/ g Extract)

M0 0.5049a ± 0.008 1.553a ± 0.004 0.2946a ± 0.026 5.39a ± 0.189 2.272a ± 0.032

M 3.0 0.7342b ± 0.012 4.205c ± 0.023 0.4488b ± 0.006 10.62b ± 0.06 3.757bc ± 0.2

M 5.0 0.8575bc ± 0.013 7.428d ± 0.041 0.5063bc ± 0.016 11.36c ± 0.16 4.193cd ± 0.007

M 7.0 1.035d ± 0.038 7.913d ± 0.069 0.5579c ± 0.015 12.82d ± 0.16 4.523d ± 0.034

M 10.0 1.2375e ± 0.037 9.012e ± 0.247 0.4908bc ± 0.005 13.48d ± 0.06 3.483b ± 0.189

M 15.0 0.8217 b± 0.008 3.363b ± 0.072 0.5521c ± 0.002 12.28e ± 0.06 4.1cd ± 0.003

Data are presented as Mean ± Standard error of mean. Values in the same column with different alphabetical superscripts are
considered statistically significant (p < 0.05). M0=100% wheat flour (control); M3.0= 3% MOLP : 97% wheat flour; M5.0= 5%
MOLP : 95% wheat flour; M7.0 = 7% MOLP : 93% wheat flour; M10.0 = 10% MOLP : 90% wheat flour; and M15.0 =15%
MOLP:85%wheatflour.
Total antioxidant is measured in terms of radical scavenging activity of antioxidant agents free radicals like DDPH

radical and by FRAP assay. The primary sources of naturally occurring antioxidants are whole grains, fruits and

vegetables. DPPH is widely used free radical to test the ability of compounds to act as free radical scavengers and to

evaluate the antioxidant activity of foods. DPPH is a stable free radical with violet colour. If free radicals have been

scavenged, DPPH will change its colour from violet to pale yellow or colourless. The results for the reducing power

potential, FRAP and DPPH sacavenging abilities varied slightly from each other. From the results, there was steady

increase in reducing power potential (figure 1) as the concentration of moringa leaves to flour increased except for 15%

MOLP bread, where a slight decrease was observed. Bread made with 10% MOLP was significantly high in reducing

power potential (3.175 ± 0.065 mg AAE / g extract) when compared to the other groups and the control (100% flour

bread; 0.294 ± 0.034 mg AAE / g extract).

There was significant (p < 0.05) increase in FRAP activities in all the bread substituted with moringa leaves when

compared to the bread made from whole wheat alone (figure 2). However, this increase in FRAP activities among the

groups is not significantly different from each other. The highest FRAP activity was observed in 7% MOLP bread

when compared to the control (100% flour bread).

The result for DPPH scavenging ability showed that composite flour exhibited enhanced antioxidant and free radical

inhibiting activity (figure 3). Just like most of the phytochemicals, highest activity was found in 10% MOLP bread
when compared to the control (100% flour bread). These results as regards FRAP and DPPH are consistent with that

reported by Supriya et al. (2015). In their study of bread (composite flour), improved antioxidants were recorded.

3.5
e
mg Ascorbic acid equivalent / g Extract

3
f
2.5 d M0
M 3.0
2
c M 5.0
1.5 b M 7.0

1 M 10.0
M 15.0
0.5 a
0
Reducing Power

Figure 1: The Reducing Power Potential of composite bread made with MOL.
Data are presented as Mean ± Standard error of mean. Values in the same column with different alphabetical
superscripts are considered statistically significant (p < 0.05). M0=100% wheat flour (control); M3.0= 3% MOLP :
97% wheat flour; M5.0= 5% MOLP : 95% wheat flour; M7.0 = 7% MOLP : 93% wheat flour; M10.0 = 10% MOLP :
90% wheat flour; and M15.0 =15% MOLP : 85% wheat flour.
 1.8
C C C
mg Ascorbic acid equivalent / g Extract

1.6 b C

1.4
a M0
1.2
M 3.0
1
M 5.0
0.8
M 7.0
0.6 M 10.0
0.4 M 15.0
0.2
0
FRAP

Figure 2: The Ferric Reducing Antioxidant Potential of composite bread made with MOLP.
Data are presented as Mean ± Standard error of mean. Values in the same column with different alphabetical
superscripts are considered statistically significant (p < 0.05). M0=100% wheat flour (control); M3.0= 3% MOLP :
97% wheat flour; M5.0= 5% MOLP : 95% wheat flour; M7.0 = 7% MOLP : 93% wheat flour; M10.0 = 10% MOLP :
90% wheat flour; and M15.0 =15% MOLP : 85% wheat flour.
 100
90
80
70
% Inhibition

60
50
40
30
20 Series1
10
0

GROUPS

M0=100% wheat flour (control); M3.0= 3% MOLP: 97% wheat flour; M5.0= 5% MOLP : 95% wheat flour; M7.0 =
7% MOLP : 93% wheat flour; M10.0 = 10% Moringa oleifera Leave Flour : 90% wheat flour; and M15.0 =15%
MOLP:85%wheatflour.MOLP=Moringa oleifera leaf powder.
Proximate analysis of the different composition flour and bread

Proximate analysis is important for evaluating the nutritional content of food products. The

different chemical composition of composite flour affects the nutritional quality of the

product. The total protein, dietary fiber, moisture, fat, ash, carbohydrate and energy content

in composite flours and final bread prepared from them are shown Table 2. From the results,

the moisture content varied significantly (p < 0.05). There was significant decrease (p <

0.05) in moisture content in 3% (11.75±0.115 %), 7% (8.92±0.06 %) and 10% (11.45±0.04

%) MOLP bread when compared with the control (12.09±0.11 %). While a significant (p <

0.05) increase was observed in 5% (13.01±0.06 %) and 15% MOLP bread (13.63±0.03 %).

Grain quality is very much depends on its moisture content. Moisture content is the indicator

of grain storability. High moisture content increases the microbial activity which

deteriorates the product during storage.

The ash is composed of non-combustible, inorganic minerals that are concentrated in the

bran layer. Ash content can be attributed to the mineral content in the sample. The result

also showed a significant (p <0.05) increase in ash content of 10% (2.47% ±0.02) and 15%

(2.43 % ±0.05) MOLP bread when compared to the control (1.84 % ± 0.02) . The result

further showed significant (p < 0.05) increase in crude protein content of all supplemented

groups. Highest protein content was seen in 15% MOLP bread (16.97±0.08 %) when

20
compared with the control (8.05±0.01). This significant increase in the protein content in the

composite flours could be attributed to high protein content in Moringa oleifera leaves.

Crude fat also revealed significant (p < 0.05) increase in all the supplemented groups, with

highest value observed in 15% MOLP (14.24% ± 0.08) when compared with the control

(10.32% ± 0.06 %). The high fat content in composite flours could have the ability to make

bread without or less addition of shortening agent.

From the results also, there was significant (p < 0.05) increase in crude fiber of all the

supplemented groups, with highest value observed in 15% (1.36% ± 0.02) when compared

to that of the control (0.65% ± 0.02). The main role of fiber is to keep the digestive system

healthy. Fiber has also been shown to benefit diabetes (Montonen et al., 2003) blood

cholesterol levels (Anderson et al., 2005), reduces constipation, coronary heart disease (Liu

et al., 1999), and obesity (Lairon et al., 2005).

The results of also showed significant decrease in the carbohydrate content of all

supplemented groups. 3% (58.26% ± 0.29), 5% (56.30% ± 0.36), 7% (58.08 % ± 0.06), 10%

(57.06 % ± 0.13) and 15% (51.37% ± 0.08) when compared to the control (67.06% ± 0.16).

The decrease observed in the carbohydrate content with increase in MOLP of bread samples

may be as a consequence of the increase in other parameters examined. These results are in

21
agreement with that of Summaya et al. (2016). In their study, significant improvement in

nutritional composition was recorded in wheat – oat composite bread when supplemented

with moringa oleifera leaf powder.

22
Table 3: Proximate Composition of Composite Bread Made from Moringa oleifera Leaf Powder.

% Composition

Samples Moisture Ash Crude Protein Crude Fat Crude Fiber Carbohydrate

M0 12.09± 0.11c 1.84±0.02b 8.05±0.01a 10.32±0.06a 0.65±0.02a 67.05±0.16e

M 3.0 11.75±0.15b 0.57±0.02a 10.84±0.14b 16.85±0.11d 0.74±0.03b 59.25±0.29d

M 5.0 13.01±0.06d 1.79±0.01b 11.49±0.46c 16.64±0.07d 0.77±0.01b 56.30±0.36b

M 7.0 8.92±0.10a 1.83±0.05b 15.66±0.02d 14.18±0.05c 1.32±0.05c 58.09±0.06d

M 10.0 11.45±0.04b 2.47±0.02c 14.66±0.12e 13.05±0.03b 1.31±0.02c 57.06±0.13c

M 15.0 13.63±0.03d 2.43±0.05c 16.97±0.08f 14.24±0.08c 1.36±0.02c 51.37±0.08a

Data are presented as Mean ± Standard error of mean. Values in the same column with different alphabetical
superscripts are considered statistically significant (p < 0.05). M0=100% wheat flour (control); M3.0= 3% MOLP :
97% wheat flour; M5.0= 5% MOLP : 95% wheat flour; M7.0 = 7% MOLP : 93% wheat flour; M10.0 = 10% MOLP :
90% wheat flour; and M15.0 =15% MOLP : 85% wheat flour.

23
CONCLUSION

Bread was produced successfully from whole wheat flour and moringa oleifera leaf powder

(MOLP). Bread produced with 7% and 10% MOLP significantly increased phytochemical and

antioxidant contents of obtained bread. Supplementation of bread with MOLP also improved on

the nutritional composition (fibre, protein, crude fat and minerals) and decreased carbohydrate

and moisture content with extended shelf life of the bread. It can be concluded from the present

study that use of the formulated composite flour can be considered in the preparation bread

enriched with physiological-functional and nutritive properties and can therefore be suggested

for incorporation as functional foods for the management of diseases like diabetes.

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