BAUCHI STATE UNIVERSITY GADAU

SEMINAR PRESENTATION
ON

TOPIC

EXTRACTION AND CHARACTERIZATION OF ESSENTIAL OIL FROM

MORINGA SEED

SUBMITTED BY

NAME ALIYU YUNUSA ABDULKADIR

MATRIC NUMBER BASUG/UG/SCI/CHM/20/10689
SCHOOL BAUCHI STATE UNIVERSITY GADAU, BAUCHI STATE

DEPARTMENT DEPARTMENT OF CHEMISTRY

COURSE CODE CHM4200
COURSE TITTLE SEMINAR PAPER
SUPERVISOR MAL. ABDULRASHID YUSUF

Sept, 2023.

TABLE OF CONTENTS
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CHAPTER ONE
1.0 INTRODUCTION………………………………………………………………………….…4
1.1 Background to the study………………………………………………………………………4
1.2 Statement of the problems………………………………………………………………….…6

1.3 Aims and Objectives of the study…………………………………………………………..…6

1.4 Research questions………………………………………………………………………….…7

1.5 Significance of the study………………………………………………………………………7

1.6 Limitations of the study………………………………………………………………….……8

CHAPTER TWO

2.0 LITERATURE REVIEW………………………………………………………………......…9

2.1 Introduction……………………………………………………………………………………9

2.2. Cultivation for Seed Production……………………………………………………………9

2.3. Flower Biology and Reproduction…………………………………………………………11

2.4. Production………………………………………………………………………………...…11

2.5. Chemical Characteristics of Seeds and Oil…………………………………………….…12

2.6 Chemical composition of Moringa oleifere seeds. ………………………………………...13

2.7 Physical and chemical characteristics of the oil 13……………………………………….…13

2.7.1 Fatty acid composition (g/100 g) of oil. ………………………………………………...…15

2.7.2 Sterol composition of Moringa oleifere oil (%) .…………………………………….…15

2.7.3 Tocopherol content (mg/kg) of Moringa oleifere oil. ………………………………….…16

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2.8 Phytochemistry and Medicinal Uses 16……………………………………………………17

2.9 Empirical Review 19…………………………………………………………………………19

2.10 Implication of the study 20…………………………………………………………………20

2.11 Uniqueness of the study………………………………………………………………….…20

CHAPTER THREE

3.0 METHODOLOGY…………………………………………………………………………21

3.1 Research design…………………………………………………………………………...…21

3.2 Area of the study……………………………………………………………………………..21

3.3 Sample preparation and Collection………………………………………………………..…21

3.4 Materials…………………………………………………………………………………..…21

3.5 Preparation Moringa oleifere seed for local extraction………………………………….…21

3.6 Methods of oil extraction…………………………………………………………………….22

3.8 methods of data analysis……………………………………………………………………..22

REFERENCES…………………………………………………………………………………23

CHAPTER ONE
1.0 INTRODUCTION

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1.4 Background to the study
Moringa oleifere, commonly referred to as "Moringa" is the most widely cultivated species

of the genus Moringa, which is the only genus in the family Moringaceae. Moringa, which is the

only genus in the family Moringaceae, is an exceptionally nutritious vegetable tree with a variety

of potential uses (Howdey, 2017). Every part of Moringa oleifere such as the seed, root and

stem are useful. The tree itself is rather slender, with drooping branches that grow to

approximately 10m in height. Moringa oleifere is called different names worldwide (Bengo,

2017). Moringa oleifere , known as “Moringa” or “Malunggay” is an Indian tree that also grows

in Asia, Africa, South- america, the Caribbean and Oceania. The oil extracted from Moringa is

known as Ben oil and reportedly con- tains 70% of oleic acid, an 18-carbon long monounsatu-

rated fatty acid (MUFA). Since the oleic acid has good oxidative stability when compared with

polyunsaturated fatty acids (PUFAs), it has found use in the food industry, as it allows for longer

storage and high-temperature frying processing. It also has uses in medicine and water treatment.

According with (Abdulkarim 2019).

Moringa oil is more stable than canola oil, soybean oil, and palm oil when used in frying.

Blending Ben oil with sunflower oil and soybean oil enhances the oxidative stability of the mix-

ture. Another potential use of Moringa oil is as biodiesel feedstock. Currently, there is a nascent

biodiesel industry in the Yucatan Peninsula, Mexico, that has been active for the last 2 - 3 years.

Several groups are trying to obtain biodiesel from vegetable oils, especially Jatropha curcas, and

from waste cooking-oil. As an alternative source of oil, Moringa seeds have been proposed as a

potential source to complement the mentioned feedstock. In this study, experimental results of

solvent Soxhlet extraction using n-hexane and ethanol, and supercritical extraction with CO2 are

reported. Solvent extraction has been reported by (Mani, 2020) using n-hexane, petro- leum ether

4
and acetone. Experimental Soxhlet extraction using n-hexane and ethanol, and also supercritical

extrac- tion with CO2 on Moringa seeds have been reported by (Nguyen,2018). Extraction of

essential oil components using supercritical fluids has received much attention in the past years

because it presents an alternative to solvent extraction or steam distillation. Sovova and Stateva,

(2020) recently review the field of supercritical extraction of vegetable materials and say that

industrial applications are increasing. It is hoped that the results will provide information to

compare Moringa seeds cultivated in Yu- catan-Mexico as raw material candidate for vegetable

oil and biodiesel.

Mani (2020) reported that comparing its chemical properties, Moringa seed oil is considered

equivalent to olive oil, and may be used for human consumption. Also, the oil from Moringa

seeds has shown the strongest anti- fungal activity against a zoophilic dematophyte caused

marked inflammatory reactions in humans Moringa oleifere is now grown widely throughout

the tropics of which Nigeria is a part (Howdey, 2017). It is sometimes known as the ‘drumstick’

or ‘horseradish’ tree ranging in height from 5 to 12m with an open, umbrella-shaped crown,

straight trunk and corky, whitish bark, the tree produces a tuberous tap root. The evergreen or

deciduous foliage (depending on climate) has leaflets 1 to 2 cm in diameter; the flowers are

white or cream coloured (Aleiro, 2017). The fruits (pods) are initially light green, slim and

tender, eventually becoming dark green, firm and up to 120cm long, depending on the variety.

Fully mature, dried seeds are round or triangular, the kernel being surrounded by a lightly

wooded shell with three papery wing. In the tropics, it is used as forage for livestock; in many

countries, Moringa is used as a micronutrient powder to treat diseases. The green pods, fresh and

dried leaves are used as vegetable. The seeds contain up to 40% of oil by weight which is used

for cooking, soap manufacture, cosmetic base and in lamps. All parts of the plant are used in a

5
variety of traditional medicines (Aleiro,2017). The press cake, obtained following oil extraction,

is useful as a soil conditioner; the plants are grown as live fences and windbreaks (Aleiro,2017).

It is also used as fuel wood source after coppicing (cutting back the main stem to encourage side

shoots); as an intercrop with other crops and the wood pulp may be used for paper-making. In the

tropics, it is used as forage for livestock; in many countries, Moringa is used as a micronutrient

powder to treat diseases. The green pods, fresh and dried leaves are used as vegetable.

1.5 Statement of the problems

Due to ever diminishing sources of fats and oils, there is the growing need for the search

of new sources of oil as well as exploiting sources that are currently unexploited in order to

supplement the existing ones. Moringa, a very rapid growing tree found growing in a varying

range of climatic condition is a promising tree and has the potential to become a new source of

oil for Nigeria. It has been reported that some 3000 kg of seeds could be obtained from one

hectare, equivalent to 900 kg oil/hectare, comparable to soybean which also yields an average

of 3000 kg seeds/hectare but with only 20% oil yield. The oil is edible, and closely resembles

olive oil in its fatty acids composition.

1.3 Aims and Objectives of the study

The aim of this research work is to find the extraction methods and characterization of essential

oil from Moringa seed

The specific objectives of this is to;

The following are objectives of this research work;

1. To extract oil from Moringa oleifere , seeds using local extraction methods.

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 2. To determine the percentage oil yield from Moringa oleifere seed.

3. To determine the density and specific gravity of Moringa oleifere seeds oil.

4. To determine the pH level of the Moringa oleifere , seed oil

5. To determine the organic septic properties of the Moringa oleifere , oil

1.4 Research questions

The following research will guide this research work,

1. Why is local extraction method is suitable Moringa oleifere ?

2. What is the percentage of oil yield of Moringa oleifere extracted using local method of

extractions?

3. What are the density and specific gravity of the local methods of extractions?

4. What is the pH level of Moringa oleifere using local method extractions?

5. Is there any organic septic properties of the Moringa oleifere oil using local extractions

methods?

1.5 Significance of the study

This research work will be beneficial the government by aware on the local extractions methods

of Moringa oleifere seed oil and provides local materials for local extractions and in return

reduce the importation of foreign oil in to the country. It will boost the morale of the farmers in

knowing the local way of extracting Moringa oleifere oil from Moringa oleifere seed. It will

makes a lot of the people to possess skills on local extractions and there by serves as means of

7
income to peoples. It will create job opportunity to community that are cultivating the Moringa

oleifere seed.

1.6 Limitations of the study

The study is limited to biology department within the biological laboratory in Aminu Saleh

College Of educations Azare Katagum Local Government Area in Bauchi State. While carrying

out the research the researcher has to consider the time and means of spending money.

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 CHAPTER TWO

LITERATURE REVIEW

2.1 Introduction

Moringa oleifere (Moringaceae) is a fast-growing softwood tree indigenous to sub-Himalayan

tracts of Northern India. It is one of 13 species within the same genus, and has become the most

diffuse in tropical and subtropical areas at altitudes up to 2000 m . Nowadays, M. oleifere is

mainly found in the Middle East and in African and Asian countries, but, due to its adaptability,

it is spreading to other areas, especially tropical and subtropical lands affected by drought. All

parts of the Moringa tree (leaves, seeds, roots and flowers) are suitable for human and animal

consumption. The leaves, which are rich in protein, minerals, β-carotene and antioxidant

compounds, are used not only for human and animal nutrition but also in traditional medicine.

The seeds, instead, have attracted scientific interest as M. oleifere seed kernels contain a

significant amount of oil (up to 40%) with a high-quality fatty acid composition (oleic acid >

70%) and, after refining, a notable resistance to oxidative degradation . The oil is known

commercially as “Ben oil” or “Behen oil”. Its properties make it suitable for both human

consumption and commercial purposes. Indeed, Moringa oil could be a good substitute for olive

oil in the diet as well as for non-food applications, like biodiesel, cosmetics, and a lubricant for

fine machinery. Moreover, after oil extraction, the seed cake can be used in waste water

treatment as a natural coagulant or as an organic fertilizer to improve agricultural productivity,

(Elenga,2020).

2.2. Cultivation for Seed Production

There are two main ways of obtaining M. oleifere plants: sowing and the use of cuttings,

(Bernat,2020). For seed production, sowing is preferred as improved varieties can be selected for

9
cultivation, ensuring proper and profitable production. Seed production, according to harvest and

management practices, (Collin,2020), requires a low density plantation (typically, 2.5 m × 2.5 m,

or 3 m × 3 m) with a triangular pattern, although 1.2 m along a row and 5 m between rows also

seems suitable for satisfactory yields ,(Collin,2020). For leaf production, the spatial distribution

in planting can vary: intensive (spacing from 10 cm × 10 cm to 20 cm × 20 cm), semi-intensive

(spacing 50 cm × 100 cm), or integrated into an agroforestry system (spacing distance of 2–4 m

between rows) ,(Elenga,2020).

Normally, Moringa seeds are sown during the rainy season and can germinate and grow without

irrigation, but for commercial purposes, irrigation through a drip system is recommended,

allowing seed production during the dry season as well. Should irrigation be employed, its

conditions depend on the cultivation area. In a recent study, (Muhl,2020).who used a rainfall

exclusion method, administered three irrigation treatments: 900, 600 and 300 mm per annum

through drip irrigation, simulating three total annual rainfall amounts. The study showed that a

restricted water supply caused no stress to the trees during the reproductive stages; moderate

water prior to floral initiation could be beneficial, stimulating flower initiation, while ample

irrigation thereafter ensured better fruit set and greater yield.

Although M. oleifere can produce a large quantity of seeds when fertilization is adequate, there

has been no exhaustive research on this issue. Several studies focused on leaves or biomass

production, suggest that the vegetative growth of Moringa is best supported by 120 kg N:P:K/ha.

Fertilization must be done during soil preparation before sowing, and when the trees are at the

onset of the growth period, i.e., just before the rainy season. Manure or compost can be used

instead of chemical fertilizer ,(Morgan,2020).

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2.3. Flower Biology and Reproduction

Moringa inflorescences are loose panicles produced in axillary, drooping 10 to 25 cm in length.

The bisexual zygomorphic flowers are up to 12 mm long and are white or cream in colour,

fragrant, and have 5 pale green sepals, 5 white petals, 5 stamens with anthers and 5 without

(staminoid),(Elenga,2020). The flowers are highly cross-pollinated due to delayed stigma

receptivity, and successful pollination requires a large number of insects ,(Morgan,2020).

The fruit is a trilobite capsule (referred to as a pod), 20–60 cm in length, and it ripens about three

months after flowering. The pods become brown and dry at maturity and split open into 3 parts

longitudinally. Each pod usually contains 12 to 35 round seeds, 1 cm in diameter. The seeds have

a brownish semi-permeable hull with three papery whitish “wings” set around it at 120-degree

intervals. A single tree can produce from 15,000 to 25,000 seeds, each weighing, on average, ca.

0.3 g. Early flowering varieties produce pods in six months, while other varieties require more

than one year ,(Muris,2020).

2.4. Production

Seed production varies tremendously. Ndubuaku (2021) studied M. oleifere yields across

Nigeria and reported 4 to 24 tons of seeds/hectare, depending on location, soil type, and

vegetation and climate conditions. There is little information about other countries’ production.

The present-day interest in M. oleifere is not its commercial value, it is its multipurpose uses

and reliability in guaranteeing good yield. Traditionally, its cultivation is almost exclusively for

fresh pod production where there is low population density . This reliability is not the case for

other crops in countries where people are often faced with famine due to crop losses, and where

whole populations are very likely to suffer nutritional deficiencies. However, before proposing

11
Moringa cultivation for such countries, studies are needed as little information is available on

Moringa seed yield under a high-density orchard design, (Ndubuaku,2021).

Recently, Ayerza, (2021) performed a comparative trial in the four ecosystems of Argentina and

Bolivia (Arid Chaco, The Yungas Tropical Forest, Sub-Humid Chaco Lowland, Tropical Forest)

to determine the seed yield and oil of Periyakulam-1 (PKM-1), an early Moringa variety

provided by the Horticultural Research Station of the Tamil Nadu Agricultural University

(TNAU) in India. Differences between years and localities were investigated. Ayerza showed

that over the three-year field trials, the oil percentages for seeds produced at Arid Chaco were

significantly higher than at Sub-Humid Chaco; however, the seed/tree yields and the oil/tree

content among the ecosystems did not differ significantly. Interestingly, when oil percentage per

tree and seed yield/tree were combined, the trees from Sub-Humid Chaco and Yungas Tropical

Forest yielded a significantly higher oil content than from the Arid Chaco trees, suggesting that

location affects yield, presumably due to one or more environmental factors such as temperature,

light, soil type and available nutrients.

2.5. Chemical Characteristics of Seeds and Oil

M. oleifere seeds are globular, about 1 cm in diameter. They are three-angled, with an average

weight of about 0.3 g, 3-winged with wings produced at the base of the seed to the apex 2–2.5

cm long, 0.4–0.7 cm wide; the kernel is responsible for 70%–75% of the seeds (A), kernels (B),

fruits (C) and oil (D) of Moringa oleifere , (Mohl,2021).

Oil is the main component of the seed and represents 36.7% of the seed weight. The oil can be

extracted almost entirely by solvent extraction, generally n-hexane, whereas less yield is

obtained by cold press extraction. In fact, only 69% (on average) of the total oil contained in

12
seeds can be extracted by cold press. Among rural dwellers, the edible oil is extracted by boiling

de-husked seeds with water, and collecting the oil from the surface of the water (Noris 2021).

Apart from the oil, the seed has a high protein content, on average 31.4%, whereas carbohydrate,

fibre and ash contents are 18.4%, 7.3% and 6.2%, respectively. Thus, the defatted seeds of M.

oleifere could provide an economical source of protein for use as a food supplement to

traditional diets to increase protein intake. Furthermore, like the protein fraction, M. oleifere

seeds have a high content of methionine and cysteine, close to that reported for milk and eggs

[24]. Therefore, they can be consumed together with legumes which are deficient in sulphur

amino acids. Moreover, M. oleifere seeds seem to be free of trypsin inhibitor and urease

activity, confirming the high protein digestibility (93%) of M. oleifere seeds.

2.6 Chemical composition of Moringa oleifere seeds.

M. oleifere oil is liquid at room temperature and golden yellow in colour. The extraction

method does not in any way affect the density and refractive index of the oil, and both are similar

to those of olive oil (bengo,2021). Instead, the smoke point is approximately 11 °C higher than

that of olive oil, suggesting a greater stability during the frying process. The oil obtained by cold

pressure extraction is higher in viscosity and acidity than that obtained by solvent-extraction.

This higher viscosity is due to the water bound in the oil during extraction, while the higher

acidity is attributed to the water added during the milling of the seeds prior to cold pressing.

Indeed, the water addition enhances the lipolytic enzyme action and prolongs the contact of the

seed (milled before cold pressing) with air and temperature. Nevertheless, the acidity of the cold-

pressed oil is generally moderate, indicative of its good resistance to hydrolysis. The iodine

number is lower than that of olive oil as Moringa oil is less unsaturated than olive oil. Finally,

13
the saponification value, regardless of the extraction method, is similar to that of olive oil,

(Bengo,2021)

2.7 Physical and chemical characteristics of the oil 13

Table 3 shows the fatty acid composition of M. oleifere oil. The saturated fatty acid content is

21.18%, with palmitic acid dominating, closely followed by behenic, stearic and arachidic acids.

The high behenic acid content is the reason why the oil is known commercially as “Ben” or

“Behen” oil. Small traces of cerotic, lignoceric, myristic, margaric and caprylic acids are also

reported in M. oleifere seed oil. The oil contains a high level of monounsaturated fatty acids, up

to an average of 76.73%. Oleic acid is the predominant fatty acid, and accounts for 73.57% of

the total fatty acids. Further monounsaturated fatty acids present in the oil are gadoleic and

palmitoleic acids. Small traces of erucic acid are reported by some studies. There is a very low

content of polyunsaturated fatty acids, on average 1.18%, and the content of linoleic and

linolenic acids is 0.76% and 0.46%, respectively. In addition, the oil’s fatty acid composition

does not seem to be particularly affected by the extraction method. Only one study reported a

small increment of the stearic and myristic acid content in solvent-extracted oil compared to oil

obtained by cold pressure (Dalot,2021). On the other hand, the agro-climatic characteristics of

the cultivation area and the M. oleifere variety cultivated could be the reason for some

differences in the fatty acid composition of the oil. Nevertheless, the present fatty acid

composition shows that M. oleifere seed oil falls in the category of high-oleic oils, and contains

a high monounsaturated to saturated fatty acids ratio (MUFA/SFA). The MUFA/SFA ratio is

characteristic of several oils, particularly olive oil, and has been associated with a reduced risk of

all-cause mortality, cardiovascular mortality, cardiovascular events, and stroke(Dalot,2021).

Therefore, M. oleifere seed oil could be an acceptable substitute for olive oil as the main dietary

14
fat in countries where the tree grows. M. oleifere seed oil has a monounsaturated fatty acid

content similar to that of olive oil, but from a nutritional point of view, a lower content of

polyunsaturated fatty acids is a limiting factor, which needs to be offset by the consumption of

alternative sources rich in polyunsaturated fatty acids. However, from a technological point of

view, the low content of polyunsaturated fatty acids ensures greater resistance and stability to

oxygen.

2.7.1 Fatty acid composition (g/100 g) of oil.

With regard to the sterol composition, M. oleifere seed oil differs in composition from olive oil

and other conventional vegetable oils used in cooking .

2.7.2 Sterol composition of Moringa oleifere oil (%).

The sterol fractions of the oil consist mainly of β-sitosterol, stigmasterol, campesterol and Δ5-

avenasterol, these accounting for 92% of the total sterols. Other sterol fractions are only present

in trace amounts. The composition of the sterols is not affected by the extraction method;

however, the degumming process involves a reduction in the sterol compounds. Other factors,

such as plant variety and agro-climatic conditions of cultivation, could affect the sterol

composition of the oil.The sterol fraction is of interest for its possible involvement in the

metabolism of cholesterol, lowering the circulating level of LDL cholesterol. Other studies also

suggest an antidiabetic potential of β-sitosterol. However, dietary results concerning the impact

of plant sterols on cardiovascular risk are still conflicting and inconclusive (Abot ,2021).M.

oleifere seed oil is characterized by a high tocopherol content, consisting of α-, γ- and δ-

tocopherols.

15
2.7.3 Tocopherol content (mg/kg) of Moringa oleifere oil.

The tocopherol content could depend on the extraction method used to obtain the oil; however,

results in this context are controversial (Bengo,2021). In fact, although one study showed the

content of α- and δ-tocopherol to be significantly higher in oil obtained by cold pressing, another

study, by the same authors, found a marked similarity in the α- and δ-tocopherol content of oils

obtained by cold pressing and solvent extraction. Nevertheless, the tocopherol content is higher

than that of other oils. The average content of α-tocopherol, which has the greatest vitamin E

potency, reaches 132.3 mg/kg, whereas the average content of the γ- and δ-tocopherols is 63.9

mg/kg and 81.2 mg/kg, respectively, also higher than contents reported for oils from seeds of

other Moringa family species. Such a high tocopherol content can be expected to contribute good

oxidative stability and protection to M. oleifere oil during its storage and processing

(Paul,2021).

Besides its use in human nutrition, M. oleifere oil also suits non-food purposes. Indeed, M.

oleifere oil has attracted the attention of researchers because of its potential in the production of

biodiesel. A recent study, (Taylor,2021).conducted in Australia reports that approximately 3030

kg of oil can produce 1000 L of biodiesel. Note that an equivalent of 3.03 tonnes/ha of oil seeds

can be harvested from dry land, and 6.06 tonnes/ha from irrigated land. Moreover biodiesel

production with Moringa seed oil is not in direct competition with existing farmlands or food

crops (second generation), and as Moringa can be grown on degraded land, studies suggest that

Moringa biodiesel is an acceptable substitute for fossil fuels, even compared to biodiesel derived

from vegetable oils of other species (Taylor,2021).

16
2.8 Phytochemistry and Medicinal Uses 16

In addition to their macronutrient composition, making them attractive for oil production and as

an economic source of protein, the seeds of M. oleifere have been extensively studied for their

content in secondary metabolites, also making them interesting for medical

purposes(Sainty,2021). Several studies have found good antioxidant activity and have isolated

phytochemical compounds that, because of their biological properties, can be used as

nutraceutical molecules . The total phenolic content of M. oleifere seeds has been found to be in

the range of 4581–4953 mg/100 g,similar to leaf amounts . The flavonoids are represented by

catechin, epicatechin, quercetin and kaempferol , present mainly in the bound forM. Moreover,

several phenolic acids have been identified, gallic acid predominating, followed by ellagic and

caffeic acids. Phenolic acids, like p-coumaric, vanillic, protocatechuic, ferulic and cinnamic

acids, have also been identified in M. oleifere seeds, but in smaller amounts. Interestingly, M.

oleifere seed also contains important bioactive compounds including alkaloids, glucosinolates,

isothiocyanates and thiocarbamates (Sainty,2021).

Folk medicine uses raw or crushed M. oleifere seeds as a decoction for treating stomach pain,

ulcers, poor vision, joint pain and for aiding digestion. The seed extract has been found to

possess good antimicrobial activity against numerous bacterial and fungal species . Many of the

phytochemical compounds isolated from the seeds are able to inhibit the growth of certain

pathogenic microorganisms responsible for human infections. For this reason, some authors have

speculated the use of these phytochemicals as an alternative to traditional therapies as they can

be pharmacologically effective with low or no side effects. The seeds’ antimicrobial activity is

also related to the presence of a short cationic protein. This protein, known as the M. oleifere

cationic protein, causes bacterial cell damage through rapid flocculation and the fusion of cell

17
inner and outer membranes. Because of their antimicrobial activity, M. oleifere seeds are used

as nature-based solutions for the problem of water purification in developing countries, using

them as an alternative to Western methods. Studies have shown that this simple filtering method

reduces not only water pollution but also harmful bacteria.

The seeds of M. oleifere have been found to be good antioxidants, able to reduce oxidative

damage associated with aging and cancer. Many of the bioactive compounds isolated from M.

oleifere seeds have been found to be potential antitumor promoters. However, recent study

results found that the ethanol extract of M. oleifere seeds had no significant effect in inhibiting

the proliferation of breast and colorectal tumour cells. Nevertheless, a recent study observed a

cytotoxic effect of M. oleifere oil in several cancer cell lines.

One study reported that M. oleifere seeds possess hepatoprotective, anti-inflammatory and anti-

fibrotic properties against CCl4-induced liver damage and fibrosis. After intoxication with CCl4,

rats simultaneously receiving treatment with Moringa seed ethanol extract (1 g/kg body weight)

for 8 weeks, compared with untreated rats, showed:

(i) lower serum levels of AST and ALT and higher serum albumin levels, indicating

better liver synthesis function.

(ii) lower globulin levels, reduced myeloperoxidase activity as well as lower hepatic

infiltration of inflammatory cells, indicating less inflammation.

(iii) lower hepatic levels of hydroxyproline and a smaller number of α-smooth muscle

actin positive cells, an activation marker of hepatic stellate cells. These cells are one

of the key cell types involved in the progression of liver fibrosis, just as they are the

principal cellular source of the excess collagen synthesis during hepatic fibrosis. Anti-

fibrotic activity also appears to be associated with the antioxidant properties of M.

18
 oleifere seed extract. This is supported by lower levels of liver malondialdehyde and

protein carbonyl, and increased activity of superoxide dismutase, which would reduce

the reactive oxygen species that play an important role in the activation of hepatic

stellate cells. Interestingly, the effects observed for the ethanolic extract of M. oleifere

seeds were, in some cases, comparable to those observed for silymarin, a drug known

to have hepato-protective and anti-fibrotic properties(Taylor,2021).

2.9 Empirical Review 19

Moringa (Moringa oleifere ) seed oil is an edible vegetable oil rich in unsaturated fatty acids.

Bragg,(2021) studied, supercritical CO2 fluid extraction method to obtain the maximum yield of

Moringa seed oil. The effects of temperature, time, and pressure, three characteristics of

extractions, on the extraction rate of Moringa seed oil were investigated by single factor test and

response surface methodological approach. The optimal process conditions of supercritical CO2

fluid extraction of Moringa seed oil were determined as extraction temperature of 45°C,

extraction time of 2.5 h, extraction pressure of 50 MPa, and CO2 flow rate of 240 L/h, resulting

in a maximum yield of 38.54%. Composition analysis shows that the extracted Moringa seed oil

is rich in unsaturated fatty acids, including oleic acid, octadecanoic acid, palmitic acid, stearic

acid, eicosanoic acid, etc. Moringa seed oil also exerted potent antioxidant activities on DPPH

and hydroxyl radicals, and its efficacy was comparable to commercial peanut oil and tea oil.

Overall, this novel extraction method of Moringa seed oil may increase its potential value and

application in the food and nutraceutical industries.

The study conducted by Wang (2021) employed supercritical CO2 fluid extraction as well as the

response surface test design on the basis of single factor experiment to investigate the three

factors of extraction, temperature, time, and pressure, on the extraction rate of Moringa seed oil.

19
The composition of Moringa seed oil was then determined. Moreover, to further understand its

potential food applications and health benefits, in vitro antioxidant activity of Moringa seed oil

was determined by four factors: DPPH free radical scavenging ability, hydroxyl free radical

scavenging ability, superoxide anion free radical scavenging ability, and total antioxidant

capacity. The present study provides a scientific basis for the further development and utilization

of Moringa and Moringa seed oil.

2.10 Implication of the study 20

The implication of all those findings to the researchers work is that it serves as road map in

finding out different strategies in extraction and characterization of essential oil from Moringa

oleifere seed.

2.11 Uniqueness of the study

The uniqueness of this study is that the researcher makes use of grinding process and heating

locally while from review of the literature the different researchers use different extraction

methods industrially.

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 CHAPTER THREE

3.0 METHODOLOGY

3.1 Research design

The research design will be adapt to this research work is experimental research design.

3.2 Area of the study

This research work will carry out in Biology laboratory of Aminu Saleh College of education

Bauchi state.

3.3 Sample preparation and Collection

The dried Moringa oleifere seed will buy from Tatari Ali market within metropolis. Half

measure will be buy but not all will be used for the experiment. The actual initial weigh is

472.69 gram will be collect for the experiment.

3.4 Materials

Materials that will use for the experiment includes Moringa oleifere seed, digital weighing

balance, Blender, Pot, wooden Stirrer, serving spoon, ponding stick

3.5 Preparation Moringa oleifere seed for local extraction

The Moringa oleifere will be sieve to obtain fresh ones. The seed will be grind using blender

and will be transfer it in to motor for ponding using pounded stick.

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3.6 Methods of oil extraction

After the pounding of the seeds, the pounded extract is going to transfer in to the pot and heate it

at ambient temperature. The ponded seed will be stirre in the pot. A hot (boiled), water will be

applied in drop not in excess using serving spoon.

3.8 methods of data analysis

The result obtained will be tabulate and calculate using formula below.

W 1−W 2
Percentage (%) = X 100
W1

Where W1 = Initial weight

And W2 = Final weight

Organoleptic effect of the oil was obtained by observing: Colour, Odour and taste of the oil

22
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