CHAPTER ONE

1.0 INTRODUCTION

1.1 BACKGROND OF THE STUDY

The water melon (Citrullus lanatus) fruit belongs to the family of cucurbitaceae. The plant
produces a special type of fruit known by botanists as a berry which has a thick rind (exocarp)
and fleshy center (mesocarp and endocarp). Water melon fruit has a smooth exterior rind, green,
juicy, sweet interior flesh (dark red). The inner rind being white or light green. This fruit
contains about 6% sugar and 90% water by weight and a lot of nutrients like carbohydrate, fat,
protein, minerals like calcium, iron, magnesium, phosphorus, potassium, and Zinc and Vitamins
like vitamin B, C and notably Vitamin A through its concentration of beta- carotene. The fruit
contains many obovate, smooth compressed seeds thickened at the margin and of a black or
yellowish white color. Watermelon plays a very important role in Africa as it is used to quench
thirst when there is shortage of water. The seed of watermelon (Citrullus lanatus) can be bruised
and rubbed up with water to form an emulsion, which can be used to cure catarrhal infections,
disorders of the bowels, urinary passage and fever. It is also being used as worm expeller, in
recent years it has been used to expel tape worm (Abiodun and Adeleke, 2010). In Nigeria, the
watermelon seed is often discarded after the juicy part has been eaten. The seed constitutes a
public nuisance because the sellers of watermelon do not dispose of these in a hygienic manner.
Therefore, this project work when successfully completed will convert waste into wealth. From
previous studies on watermelon seed oil, it is light penetrating and rich in essential fatty acids
(Abiodun and Adeleke, 2010). As a result of importance of drying techniques, this research was
designed to observe the effect of two drying methods on the physico-chemical properties of the
seed oil and also to determine its edibility and effectiveness in soap making. In Africa,
Watermelon seeds have been prized for the highly nutritive oil that they contain. Traditionally,
the seeds are removed from the rind and then allowed to dry outside in the sun. Once dried, the
seeds are then pressed to extract the beneficial oil. Natural Sourcing provides the most superior
watermelon seed oil available. It is carefully processed and packaged to maintain the purity,
freshness and beneficial properties of this exceptional watermelon seed oil. The fatty acid profile
of edible oils plays an important role in their stability and nutritional value. Monounsaturated
(18:1) and polyunsaturated (18:2) fatty acids have been found to be effective replacements for
saturates as part of cholesterol-lowering diets (Mattson and Grundy, 1985).

However, it is also known that the oils with substantial amounts of unsaturation, particularly 18:2
fatty acids, are susceptible to oxidation and may produce products that contribute to
arteriosclerosis and carcinogenesis. Some studies with experimental animals indicate that
excessive amounts of linoleic acid promote carcinogenesis(Kubow, 1990).Watermelon seed oil,
rich in linoleic acid (~64.5 %), is used for frying and cooking in some African and Middle
Eastern American countries owning to its unique flavour (Akoh and Nwosu, 1992).

1
 Much research has been published on the oxidative stability of some vegetable or fruit oils, but a
little has been reported on the stability of watermelon seed oil. The modification of watermelon
seed oil fatty acid composition by incorporation of oleic acid (18:1) has been explored
(Charment et al., 1997). The modified watermelon seed oil was produced with the better balance
of monounsaturated (18:1) and essential Fatty acids (18:2), and also improved the seed oil
oxidative stability and nutritional value (Charment et al., 1998). The watermelon family
Cucurbitaceae is the most popular fruit in Serbia, with a traditional name ″lubenica″.
Unfortunately, there is no information of domestic application or medicinal values of this seed
oil for cooking and or frying as useful source of oil with good nutritional value. Thus,
watermelon seed oil composition (physical and chemical) shall be evaluated in this present study.
The data generated in the work will help in the future selection of watermelon seed oil for use in
human diet. The light texture, moisturizing capabilities and stable shelf life of watermelon seed
oil makes it a highly suitable emollient in natural baby care formulations and light body
emulsions. Unlike mineral oil that is a common ingredient in commercial skin projects,
watermelon seed oil does not clog pores or prevent the body from natural elimination of toxins
through the skin. The rich composition of essential fatty acid contained in watermelon seed oil
nourishes and restores elasticity to the skin (Akoh and Nwosu, 1992). It may be recommended
for use in skin care formulations for all skin types including dry, oily and maturing skin.
Watermelon seed oil may also be a perfect choice for inclusion in hair care formulations as it is
non greasy yet highly moisturizing. This research work will study the different physiochemical
compositions of the oil and its edibility for domestic use and in the cosmetic industry.

2
1.2 PROBLEM STATEMENT

In Nigeria, the watermelon seed is often discarded after the juicy part has been eaten. The
seed constitutes a public nuisance because the sellers of watermelon do not dispose of
these in a hygienic way. Therefore, this project work when completed will successfully
convert waste to wealth which is a good form of waste management.

Investigation indicates that a cholesterol rich diet causes abnormally high levels of
cholesterol and the related fats and lipids in the blood. Evidence strongly indicates that
people with such high levels are more likely to develop atherosclerosis and heart attacks
than those with lower levels.

1.3 AIMS AND OBJECTIVES

1.3.1 AIM

The aim of this research project is to investigate the effect of the two drying methods on
the physiochemical properties of edible water melon seed oil.

1.3.2 OBJECTIVES

1. To extract oil from watermelon seed using Soxhlet apparatus.

2. To use food grade ethanol as a solvent.
3. To carryout physiochemical analysis of the extracted oil
4. To carryout GC-MS analysis of the extracted oil
5. To study the effects of different drying methods
1.4 SCOPE OF THE STUDY
1. Extraction of oil from watermelon seed by solvent extraction method
2. Determination of the physiochemical properties of the oil
3. Determination of the effect of different drying methods on the physiochemical
properties of the oil.
4. To carryout GC-MS analysis of the extracted oil.

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 1.5 JUSTIFICATION AND SIGNIFICANCE OF THE STUDY

Nutrients in watermelon seeds are able to ward off cancer, improve and or prevent
cardiovascular disease, hypertension, and reduce levels of bad cholesterol. The oil obtained from
the seed is a good source of healthy fat high in unsaturated fat better than low-fat foods,
because of increasing HDL (good cholesterol) and lower LDL (bad cholesterol).

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 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 GENERAL OVERVIEW ON WATERMELON SEED OIL

Watermelon seed (citrullus lanatus) oil is also known as Ootanga and Kalahari oil. It is an
annual training herb which is grown widely in warm temperature, subtropical regions all over the
world. It comes from the diminutive form of citrus and referring to colour and shape of the fruit,
and vulgaris meaning common or ordinary fruit. Its English common name, watermelon comes
from the flesh of this succulent fruit, which contain over 90 % water. Native of Africa, it was a
valuable and portable source of water for desert situations and when natural water supplies were
contaminated. It is eaten fresh or raw in this part of the world for its cool and refreshing juice,
and the seeds are usually thrown away as waste. The protein content of the seeds varies from 25 -
32 % while the oil content varies from 20-50 % (Ige et al., 1984). In some parts of the world, the
seeds are eaten as sources of protein and oil, and at times ground into flour, which is
incorporated into cereal flour for bread making. Watermelons were first cultivated in Egypt
where testaments to their legacy were recorded in hieroglyphics painted on building walls. The
fruit was held with such high regards that it was placed in the tombs of many Egyptian kings. It
is not surprising that watermelon played an important role in Egypt, and subsequently in
countries in the Mediterranean region, since water was often in short supply in these areas, and
people could depend upon watermelon for its thirst-quenching properties. Watermelons were
brought to China around the 10th century and then to the Western Hemisphere shortly after the
discovery of the New World. In Russia, where much of the commercial supplies of watermelons
are grown, there is a popular wine made from this fruit. In addition to Russia, the leading
commercial growers of watermelon include China, Turkey, Iran and the United States (Ali et al.,
2008).

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In Nigeria, traditional protein foods such as meat, fish, poultry and dairy products are expensive
and therefore the majority of population cannot afford them. In this regard, attention has been
shifted to alternative sources of protein particularly among the oil seeds, kernels and less known
plant sources. Extraction and characterization of a food source, determine not only average
nutrient content, but also help to find alternative uses for such food materials.

2.2 Oil

2.2.1 Fatty Acids

The fatty acid (FA) composition determines the physical properties, the stability and the
nutritional value of lipids. All lipids of natural origin consist of saturated FA, monoenoic FA, and
polyunsaturated fatty acids (PUFA) in various proportions and differ in detailed FA composition.
Variations in plant and animal lipid FA composition make it possible to determine the origin of
the lipids. FA distribution in triacylglycerol (TAG) as well as in phospholipids affects the physical
properties, lipolytic and oxidative stability, and nutritional availability of lipids. In many TAGs,
the fatty acids are arranged in a non-random distribution. In plants, monoenoic FA and PUFA
are dominant at sn-2 position. In some animal fats the TAG sn-2 position is occupied by palmitic
acid (Kolakowska et al., 2003).

2.2.2 Tocopherols

The tocochromoanols comprise a group of chemically related tocopherols and tocotrienols,

which are similar in molecular structure and occur in plants, plant oils, nuts, grains, fruits and

vegetables. All compounds in the homologues, exhibit antioxidative and vitamin E activity. In all

6
homologues the basic structural unit is a chroman ring system (2-methyl-6-hydroxy-chroman)

with an isoprenoid side chain with 16 C-atoms. The homologues differ in the number and

distribution of methyl groups in the aromatic ring. Tocopherols differ from tocotrienols because

they have a saturated side chain. Tocopherols are antioxidants possessing a “carry through”

property; this is defined as the ability of an antioxidant to survive the technological process,

mainly the heat treatment of food stuffs, and transfer the stabilizing activity to the final

product. Acting as chain-breaking antioxidants, tocopherols react with lipid radicals to convert

them into more stable products (Kolakowska et al., 2003).

The major lipid radical formed at normal oxygen pressure is the peroxy radical ROO, which is

converted into a hydroxyperoxide by a hydrogen donator. The mode of action of tocopherols as

antioxidants is explained by their role in donating hydrogen from their phenolic group to

radicals, to stabilize them and to stop the growing formation of hydroperoxides. The resulting

tocopheryl semiquinone radical molecule itself, which loses the antioxidative properties, shows

different possibilities for further reacting. Two tocopheryl semiquinone molecules can form one

molecule of the stable tocopherylquinone and one regenerated molecule of tocopherol. The

reaction between the two radicals may also form tocopherol dimers, especially the g -

tocopherol-biphenyl-dimer and the g-tocopherol-ether-dimer, which have antioxidative

properties. A model study showed that g -tocopherol was superior to a-tocopherol as an

antioxidant, because it oxidizes to more stable compounds, which are still effective as

antioxidants (Wagner and Elmadfa, 1999).

7
The tocopherol content of foods is important to protect food lipids against autoxidation and,
thereby to increase their storage life and their value as wholesome foods.

2.2.3 Oxidative Stability

Oxidative stability is an important parameter for evaluating the quality of oils and fats, as it

gives a good estimation of their susceptibility to oxidative deterioration, the main cause of their

alteration. Lipid oxidation has been recognized since antiquity as a development of quality

deterioration of fats and oils, resulting in the development of unpleasant taste and odours as

well as changes in colour, viscosity, specific gravity and solubility of products. Several authors

have reviewed the mechanisms involved in lipid oxidation and its subsequent effects in foods.

Autoxidation is a natural process that takes place between molecular oxygen and unsaturated

fatty acids. Autoxidation of unsaturated fatty acids occurs as a free radical process that consists

of three basic steps of initiation, propagation and termination. Initiation starts with the

abstraction of a hydrogen atom adjacent to a double bond in a fatty acid (RH) and this may be

catalyzed by light, heat or metal ions to form a free radical (Shahidi, 1995).

2.2.4 Refining

The aim of refining is to achieve the maximum attainable elimination of all non- triacylglycerol

or partial acylglycerol components of crude oils with the maximum saving of unaltered

triacylglycerols, natural antioxidants and vitamins. The components to be removed are thus

primarily the free fatty acids, the phosphoacylglycerols, sterols, pigments, glucosides, waxes,

hydrocarbons, and all those glyceridic and non-glyceridic lipid transformation products which

8
together with contaminations may be detrimental to the flavor or oxidative stability of the

refined oil. The refining operation consists of four steps; the sequence of these steps is:

Degumming, Deacidification (by caustic neutralization), Bleaching or decolorization,

Deodorization (Hoffmann, 1999). Differences in the normal composition of crude oils and fats

affect the further processing conditions necessary to purify the oil. Not all the co-constituents

in the crude oils are undesirable. The sterols are colorless and heat stable and, for all practical

purposes, inert; hence they pass unnoticed unless present in unusually large amounts.

Tocopherols have the important function of protecting the oil from oxidation. Most of the

other compounds are objectionable constituents, since they render the oil dark colored, cause

it to foam or smoke, or are precipitated when the oil is heated in subsequent processing

operations (Hui, 1996). During the refining process fatty substances can be structurally altered

by various chemical reactions like hydrolysis, oxidation, dehydration, dimerization,

polymerization and interesterification. The type of these chemical reactions depends on the

process used for the refining process (DGF 125 Deutsche Gesellschaft für Fettwissenschaft

(German Society for Fat Science)).

Alkali refining removes free fatty acids to produce edible fats and oils with very low acid values,

even though free fatty acids are not objectionable, unless present in higher amounts. Free fatty

acids can be easily removed by washing crude fat or oil with a solution of sodium hydroxide or

sodium carbonate. Alkali refining alone improves the product quality, but it is not sufficient to

remove all objectionable substances so that other refining steps had to be introduced.

9
Phospholipids and other polar lipids are removed by alkali refining with free fatty acids, but it

was soon found that it would be better to remove phospholipids in a separate step consisting

of heating the crude raw material with water or an aqueous solution of phosphoric acid. The

sedimented materials, called gums, can be removed by centrifuging. Pigments are removed by

bleaching with bleaching clay or charcoal, and volatile substances are removed by distillation

with steam at high temperatures and under reduced pressure (deodorization). The resulting

product is nearly colourless, bland with no appreciable flavour, and has a good stability on

storage. Alkali refining has several disadvantages: The procedure is rather expensive and Loss

of neutral triacylglycerols.

In physical refining, free fatty acids are removed from crude oils by physical methods without

the application of sodium hydroxide or sodium carbonate (Dahlke and Eichelsbacher, 1998).

Several physical procedures have been developed; most of them are only of historical interest.

Steam refining is the only large scale practicable method used in industry nowadays, and it is

connected with efficient degumming and bleaching (Stage, 1983). The oxidative stability of

vegetable oils depends on the fatty acid composition and the presence of minor components

such as tocopherols, carotenoids, chlorophylls, metal ions, phosphorus and the initial amount

of hydroperoxides. Crude vegetable oils are usually oxidatively more stable than the

corresponding refined oils (Pekkarinen and Heinonen, 1998).

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2.2.4.1 Degumming

Most important degumming methods are water degumming, which is defined as treatment of

crude oil with hot water, and acid degumming, defined as treatment of crude oil with

phosphoric acid or citric acid. It is relatively easy to remove hydratable phospholipids from

crude oils as they readily absorb water and thereby becoming insoluble in oil. A major problem

with this method is the removal of non-hydratable phospholipids, mainly calcium and

magnesium salts of phosphatidyl ethanolamine. Water degumming may be sufficient for some

types of vegetable oils (watermelon seed oil), and 99 % of phospholipids are removed by

heating oil to 80–85 °C with water and then filtering at 55–60 °C. However, water degumming

does not destroy non hydratable salts of phosphatidic acids; therefore, it is not sufficiently

efficient in most cases for the preliminary purification of oils before physical refining. For acid

degumming, originally phosphoric acid was used as it forms non-dissociable phosphates with

calcium, iron, or magnesium ions and is much cheaper than polyvalent organic acids. The use of

0.05–0.20 % phosphoric acid was found satisfactory for the degumming of palm oil (Faessler,

1998) but not for most other vegetable oils (Iwuoha et al., 1996). For the degumming of

watermelon seed oil the application of phosphoric acid and water was recommended, first with

concentrated acid and then with dilute acid. Citric acid is an alternative to phosphoric acid. It is

more expensive than phosphoric acid, but the removal of non hydratable phospholipids is more

efficient. Acetic anhydride was found similarly efficient as citric acid for the degumming of

watermelon seed oil. Sodium citrate was reported as more efficient for removing calcium,

11
magnesium, or iron from non-hydratable phosphatides than free citric acid (Rohdenburg and

Perédi, 1993).

Acid degumming processes usually lead to lower residual phosphorus and iron contents, than

those of water degumming.

2.2.4.2 Caustic Neutralization

Free fatty acids can be easily removed by washing crude fat or oil with a solution of sodium

hydroxide or sodium carbonate; the process is called alkali refining. Alkali refining alone

improves the product quality, but it is not sufficient to remove all objectionable substances so

that other refining steps had to be introduced (Cmolik and Pokorny, 2000). The caustic is

prepared by dilution of sodium hydroxide with softened water to a specific gravity of about 17-

18 Baume. The amount of alkali solution added to the oil is known as the "treat," which can be

measured based on the percent FFA in the degummed oil. The correct excess treat will enable

the highest efficiency and allow a minimum loss of neutral oil. The excess treat depends on the

quality of the feedstock being alkali refined and must be constantly adjusted (Hui, 1996). For

physical refining first an acid pretreatment, (usually 0.05 to 0.2 % of phosphoric acid well mixed

at about 100 °C), takes place followed immediately by heavy conventional adsorptive bleaching

using up to 2-3 % clay. Alkali refining works on the entire undesirable, non-triglyceride

components except carotene and sulfur, and both of these are easily removed in the

deodorization step. Alkali refining is the only step other than deodorization that removes free

fatty acids (Wan, 1991).

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2.2.4.3 Bleaching

Bleaching of edible fats and oils is usually conducted at a temperature of 90 to 100 °C. For

difficult-to-bleach oils the temperature can even reach 120 °C; this temperature used during

deodorization cannot be applied to the bleaching step. Activated bleaching earth has not only

an adsorptive effect but acts also as a solid acid, a cation exchanger and catalyst of various

reactions (DGF: 135 of the Deutsche Gesellschaft für Fettwissenschaft (German Society for Fat

Science)). Bleaching is a technological process during refining of edible fats and oils, which

cannot be discussed without consideration of other processes during refining as degumming,

neutralization and deodorization. All these processes performed within the refining have to be

used to remove certain naturally occurring or secondary by-products as well as possible

contaminants, which have a negative influence on organoleptic properties, shelf life and the

use of edible oils for human consumption. A major task of the bleaching step is the

decomposition of possibly existing hydroperoxides in order to improve the stability of the

refined product. (DGF 125 of the Deutsche Gesellschaft für Fettwissenschaft (German Society

for Fat Science)). At the bleaching step due to the low temperature of 90 to 120 °C only a small

quantity of non-conjugated isomeric trans fatty acids is being formed. In the presence of an

acidic bleaching earth the formation of steradienes and disterylethers is catalytically supported

already at temperatures below 100 °C (Wan, 1991).

2.2.4.4 Deodorization process

Deodorization is the last major processing step in refining of edible oils. It is responsible for

removing both the undesirable ingredients occurring in natural fats and oils, and those which

13
may be impacted by prior unit processes such as caustic refining, bleaching, hydrogenation, or

even storage. Experience has shown that flavour and odour removal correlates well with the

reduction in free fatty acid content of the oil. Deodorization consists of steam-stripping the oil

for free fatty acids removal, which depends on stripping steam rate, pressure and temperature

(Wan, 1991). Flavour and oxidative stability of vegetable oils are usually improved by

deodorization; a comparison of flavour and oxidative stability of soybean oil deodorized at 240,

270 and 295 °C showed that oils deodorized at higher temperatures were less stable. When a

lower temperature is used, more stable oils are obtained when the deodorization period is

lengthened; when a high temperature is used, flavors and oxidative stable oils are more likely

attained when the deodorization period is short (Moser et al., 1966). Linoleic acid is on the one

hand stable at 230 or 240 °C, on the other hand at 250 °C and particularly at 270 °C the

formation of trans isomers of linoleic acid is accelerated. Linolenic acid is much less stable on

heating than linoleic acid even at low deodorization temperatures. The major factors affecting

the formation of trans fatty acids during deodorization are temperature, heating time, pressure,

and stripping steam dosage. A deodorization temperature of 220-230 °C seems to be the critical

point above which 18:3 fatty acid isomerization substantially increases. The critical temperature

is higher for 18:2 fatty acids, over 240 °C. Chemical refining should be performed at 230-235 °C

and physical refining at 235-240 °C to obtain deodorized oils with a low level of Trans FA (< 1 %)

(Tasan and Demiric, 2003).

2.3 Nutritional Benefits of Watermelon Seed Oil

14
Watermelon seed oil is highly nutritious which makes it a strong and a perfect ingredient for

baby oils and other facial oils. The light texture and moisturizing property of watermelon oil

makes it a perfect addition to natural baby oil formulations.

The watermelon seed oil does not block the pores, thus letting the body to eliminate all the

toxins naturally through the pores, saving the person from the risk of any skin infection

(Shadrach and Oyebiodun, 1999).

Watermelon seed oil is a good absorbent, making it a perfect oil to be used by people with oily

or greasy skin. It can, however, be suitable and beneficial for the other skin types as well. It is

used as an ingredient for skin care products for all the skin types including dry skin, oily skin,

acne-prone or maturing skin, because of it being rich in essential fatty acids, which helps in

maintaining the elasticity of the skin (Shadrach and Oyebiodun, 1999).

Watermelon seed oil also acts as a good cleanser, as the rich omega acid and linoleic acid within

the oil helps in the extraction of excess sebum within the skin, ensuring the cleanliness of the

skin. It can also be used to dissolve the oils that get accumulated in the pores, making it the

best option for oily skin types (Shadrach and Oyebiodun, 1999).

The mild aroma, indefinite shelf life and viscosity make it all-purpose oil. The watermelon seed

oil is also used in aromatherapy. Its perfume soothes the mind and body of a person and can be

used as an effective stress buster.

Watermelon seed oil, similar to watermelon, is resilient in nature. The fatty acid contents in the

oil contain water soluble properties that help the oil to penetrate through the skin easily,

15
providing good moisturizing effect to the skin. This moisturizing property of the oil can also be

used to keep the hair and scalp healthy and pigmented. This property makes watermelon seed

oil an effective ingredient to be used in different moisturizing and hair care products.

Watermelon oil also serves as an anti-aging agent because of its moisturizing property. It helps

lock the moisture within the skin and helps the skin to combat aging degradation, in the

process regenerating the elasticity of the skin.

2.3.1 Application of Watermelon Seed Oil

Watermelon seed oil is used in baby formulations, creams, lotions, soaps, and eye creams. This

oil is good for balancing oil production, while being restorative and nourishing in action. The

neutrality of watermelon oil makes it a perfect oil to blend for body or facial oils being effective

on greasy skin types. The light texture, moisturizing capabilities and stable shelf life of

watermelon seed oil makes it to be a highly suitable addition to natural baby oil formulations.

Unlike mineral oil that is a common ingredient in commercial baby oils, watermelon seed oil

does not clog pores and eventually helps the body to naturally eliminate toxins through the

skin. Watermelon Seed Oil is highly nourishing yet it is light oil with good absorption properties.

It is a good choice for use with oily skin but can be effective with all skin types. Hence Water

Melon seed oil is added to skin care formulations for all skin types, including dry, oily, acne-

prone or maturing skin as it is rich in essential fatty acids which help to restore elasticity to the

skin. The high content of omega acids and Linoleic acid within the oil assist in the removal of

16
excess sebum within the skin ensuring the skin is cleansed. The oil from the watermelon seeds

can be used to dissolve the oil build up in the pores. So it is best for oily skin. Watermelon seed

oil is also a perfect choice for inclusion in hair care formulations as it is non-greasy yet highly

moisturizing. The nutritious contents of the melon seed oil helps to add nourishment to

improve the strength of the hair. In addition, watermelon seed oil’s viscosity, mild aroma and

indefinite shelf life make it good all-purpose carrier oil for use in aromatherapy. Watermelon is

rich in carotenoids. Some of the carotenoids in watermelon include lycopene, phytofluene,

phytoene, beta-carotene,lutein, and neurosporene. Lycopene makes up the majority of the

carotenoids in watermelon. The carotenoid content varies depending on the variety of the

watermelon. Depending on the variety, carotenoid content in red fleshed watermelon varies

from 37 – 121 mg/kg fresh weight, where as lycopene varies from 35 – 112 mg/kg fresh weight.

Carotenoids have antioxidant activity, free-radical scavenging property. Several researches have

reported an association between dietary lycopene consumption and lower incidence in diseases

such as prostate and oral cancers. Lycopene may also help reduce risks of cardiovascular

disease. Watermelon seeds are excellent sources of (both essential and non-essential amino

acids) and oil. Watermelon seed is about 35 % protein, 50 % oil, and 5 % dietary fibr .

Watermelon seed is also rich in micro- and macro-nutrients such as magnesium, calcium,

potassium, iron, phosphorous, zinc etc. Besides, the red flesh, in the Arab world, salted and

roasted watermelon seeds are also consumed as a snack (Michigan, 2009).

17
2.3.2 Health Benefits of Watermelon

Studies suggest that these potent antioxidants watermelon may have protective effects against

heart disease and certain cancers, such as prostate, bladder, and cervical cancer. Lycopene and

beta-carotene are compounds called carotenoids, which are highly colored pigments that help

protect plants against damage from sunlight. Carotenoids are important to humans because

they have antioxidant activity and prevent free radicals from causing harm to the body, similar

to other types of antioxidants such as selenium and vitamins C and E. Some carotenoids are

also converted to vitamin A, which is necessary for normal immune function and development

of cells. Other significant carotenoids include lutein and zeaxanthin, which play a role in visual

function. The watermelon juice diet increase blood concentrations of lycopene and beta-

carotene almost five-fold and two-fold, respectively, compared with the diet without

watermelon juice. However, there was no significant increase in lycopene or beta-carotene

levels when the amount of watermelon juice was doubled. Consumption of tomato juice

produced a similar increase in blood lycopene concentrations, but failed to increase beta-

carotene levels. Although the current study shows watermelon juice increases lycopene and

18
beta-carotene blood levels, the long-term health benefits consuming watermelon have yet to

be investigated. (Kamel et al., 1982).

2.4 Leaching

Liquid can be extracted from solid by leaching. As the name implies, contacting the solid with a

suitable solvent leaches the soluble liquid contained in a solid out. The principle of application

of leaching is in the extraction of valuable oils from nuts and seeds such as palm oil and grape

seed oil. The equipment used to contact the solids with the solvent is usually designed to suit

the type of solid being processed and is to an extent unique to the particular industry.

2.4.1Methods of Extraction

2.4.1.1 Supercritical fluid extraction (SFE)

The state of a substance is called supercritical fluid when both temperature and pressure

exceed the critical point values. Extractions with supercritical fluid solvents have emerged in

recent years as highly promising environmentally benign technologies for the production of

natural extracts with high potency of active ingredients, such as flavours, fragrances, spice oils

and oleoresins (Michigan, 2009). SFE is the process of separating one component (the

extractant) from another (the matrix) using supercritical fluids as the extracting solvent.

Extraction is usually from a solid matrix, but can also be from liquids. SFE can be used as a

19
sample preparation step for analytical purposes, or on a larger scale to either strip unwanted

material from a product (e.g. decaffeination) or collect a desired product (e.g. essential oils).

Carbon dioxide (CO2) is the most used supercritical fluid, sometimes modified by co-solvents

such as ethanol or methanol. Extraction conditions for supercritical CO2 are above the critical

temperature of 31 oC and critical pressure of 74 bar. SFE is an alternative to liquid extraction

using solvents such as hexane, petroleum ether or dichloromethane. There will always be some

residual solvent left in the extract and matrix, and there is always some level of environmental

contamination from their use. In contrast, carbon dioxide is easy to remove simply by reducing

the pressure, leaving almost no trace. Extraction using supercritical fluid yields oil of high purity

however, requirement for high pressures increases the cost compared to conventional liquid

extraction, so SFE will only be used where there are significant advantages (Michigan, 2009).

2.4.1.2 Solvent extraction

This is a process which involves extracting oil from oil-bearing materials by treating it with a low

boiler solvent as opposed to extracting the oils by mechanical pressing methods (such as

expellers, hydraulic presses etc). The solvent extraction method recovers almost all the oils and

leaves behind only 0.5 – 0.7 % residual oil in the raw material while in the case of mechanical

pressing the residual oil left in the oil cake may be anywhere from 6 – 18 % (Michigan, 2009).

The solvent extraction method can be applied directly to any low oil content raw materials. It

can also be used to extract pre-pressed oil cakes obtained from high oil content materials. As a

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result of high percentage of recovered oil, solvent extraction has become the most popular

method of extraction of oils and fats. Fat is one of the essential components of the human diet,

therefore the demand for oils and fats are increasing with the increase in population and

standards of living.

Solvent extraction is basically a process of diffusion of a solvent into oil-bearing cells of the raw

material resulting in a solution of the oil in solvent. Various solvents can be used for extraction.

In a nutshell, the extraction process consists of treating the raw material with a solvent and

recovering the oil by distillation of the resulting solution of oil in the solvent called miscella.

Evaporation and condensation from the distillation of miscella recovers the solvent absorbed in

the material. The solvent thus recovered is reused for extraction. The low boiling point of

extractive solvent and the high solubility of oils and fats in it are the properties exploited in the

solvent extraction process (Michigan, 2009). It has a disadvantage for highly volatile and

flammable solvents that could result to a risk of fire and explosion.

2.4.1.3 Mechanical Pressing

Oil milling (mechanical extraction) can be only pre-pressing in which case seeds are lightly

pressed, and which leaves about 14 – 18 % oil in the pressed cake. This method results in

higher capacity, lower power consumption, lower wear and tear or maintenance. But it also

lowers recovery of oil, it is recommended when the buyer has his own solvent extraction line,

so that balance oil is recovered during solvent extraction process.

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2.4.2 Factors Influencing the Rate of Extraction

The selection of the equipment for an extraction process is influenced by the factors which are

responsible for limiting the extraction rate. Thus, if the diffusion of the solute through the

porous structure of the residual solids is the controlling factor, the material should be of small

size so that the distance the solute has to travel is small. On the other hand, if diffusion of the

solute from the surface of the particles to the bulk of the solution is the controlling factor, a

high degree of agitation of the fluid is required. There are four important factors to be

considered: (Coulson and Richardson, 2002).

2.4.2.1 Particle size

Particle size influences the extraction rate in a number of ways. The smaller the size, the

greater is the interfacial area between the solid and liquid, and therefore the higher is the rate

of transfer of material and the smaller is the distance the solute must diffuse within the solid as

already indicated. On the other hand, the surface may not be so effectively used with a very

fine material if circulation of the liquid is impeded, and separation of the particles from the

liquid and drainage of the solid residue are made more difficult. It is generally desirable that the

range of particle size should be small so that each particle requires approximately the same

time for extraction and, in particular, the production of a large amount of fine material should

be avoided as this may wedge in the interstices of the larger particles and impede the flow of

the solvent (Coulson and Richardson, 2002).

2.4.2.2 Solvent

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The liquid chosen should be a good selective solvent and its viscosity should be sufficiently low

for it to circulate freely. Generally, a relatively pure solvent will be used initially, although as the

extraction proceeds the concentration of solute will increase and the rate of extraction will

progressively decrease, first because the concentration gradient will be reduced, and secondly

because the solution will generally become more viscous.

2.4.2.3 Temperature

In most cases, the solubility of the material which is being extracted will increase with

temperature to give a higher rate of extraction. Further, the diffusion coefficient will be

expected to increase with rise in temperature and this will also improve the rate of extraction.

In some cases, the upper limit of temperature is determined by secondary considerations, such

as, for example, the necessity to avoid enzyme action during the extraction of sugar (Coulson

and Richardson, 2002).

2.4.2.4 Agitation of the fluid

Agitation of the solvent is important because this increases the eddy diffusion and therefore

the transfer of material from the surface of the particles to the bulk of the solution, as

discussed in the following section. Further, agitation of suspensions of fine particles prevents

sedimentation and more effective use is made of the interfacial surface (Coulson and

Richardson, 2002).

2.4.3 Choice of Solvent

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The choice of solvent is the most important factor to be considered for effective, complete and

economical extraction of desirable solute. The following characteristics must be accessed in the

choice of solvent for extraction (Onyeike and Acheru, 2002).

2.4.3.1 Selectivity

This is the ability to extract a substance from the mixture of two substances preferentially so

that the ratio of the extracted substance to that of the other substance in the mixture after the

removal of solvent is different from the ratio of these components in the solvent free raffinate.

2.4.3.2 Recoverability

This is the ability to recover the solvent for other uses and this must ordinarily be done by

another mass transfer operation by the method of distribution.

2.4.3.3 Volatility

This is the ability of a liquid to change into vapour or gas because the recovery of solvent is

usually by distillation.

2.4.3.4 Purity

This means that the solvent should be pure and not contain any impurity of any form so as not

to interfere with the use for which the solvent is intended.

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2.4.3.5 Solubility

This is the ability of solvent to dissolve a wide variety of substance. In solvent extraction a

solvent of high solubility power is chosen.

2.4.3.6 Inflammability

This is a measure of ease with which solvents ignite, therefore in order to avoid fire or

explosion solvents with flash point higher than 32.2 o C should be chosen.

2.4.3.7 Safety

If possible, and all other factors considered a solvent should be chosen that is not toxic or

dangerously inflammable.

2.5 Physio - Chemical Properties of Oil

2.5.1 Acid value (free fatty acid)

The free fatty acids in oil are estimated by titrating against potassium hydroxide (KOH) using

phenolphthalein. The acid value is the amount of milligram of KOH required to neutralize the

free fatty acids present in one gram of sample. It is expressed as oleic acid equivalent (Shahidi,

1995). Mohammed (2004) reported acid value as 0.53 ± 0.25 mg KOH/g and the concentration

of free fatty acid as 0.27 ± 0.13 % for pumpkin seed kernel oil, Marjana (2005) reported acid

value as 1.00 mgKOH/g and free fatty acid as 0.521(% oleic acid) for melon seed oil, Taiwo et al.

(2008) reported acid value as 13.40 mgKOH/g and free fatty acid as 6.24 mgKOH/g for

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watermelon seed oil, Fatimah et al. (2011) reported acid value as 1.57 mgKOH/g and free fatty

acid as 0.785 (% oleic acid) for winter melon seed oil, Ziyada and Elhussein (2008) reported acid

value as 2.31 mgKOH/g and free fatty acid as 1.16 mgKOH/g for colocynthoide seed oil.

2.5.2 Saponification value

This is the amount of alkali in milligram required to neutralize a definite quantity (1g) of an oil

or fat. This value is useful for a comparative study of the fatty acid chain length in oil or fat. A

known quantity of oil is refluxed with an excess amount of alcoholic KOH. After saponification

the remaining KOH is estimated by titrating it against a standard acid (Taiwo et al., 2008).

Mohammed (2004) reported saponification value as 185.20 ± 5.84 mgKOH/g for pumpkin seed

kernel oil, Marjana (2005) reported saponification value as 188 mgKOH/g for melon seed oil,

(Taiwo et al.,2008) reported saponification value as 117.81 mgKOH/g for watermelon seed oil,

Fatimah et al. (2011) reported saponification value as 182.3-194.1 mg KOH/g for winter melon

seed oil, Ziyada and Elhussein (2008) reported saponification value as 189 – 201 mgKOH/g for

colocynthoide seed oil.

2.5.3 Iodine value

It is a measure of the degree of unsaturation in oil. It is constant for particular oil or fat. Iodine

value is a useful parameter in studying oxidative rancidity of oils. Oil contains both saturated

and unsaturated fatty acids. Iodine gets incorporated into the fatty acid chain wherever the

double bonds exist. Hence the measure of iodine absorbed by oil gives the degree of

unsaturation. Iodine value or number is defined as the gram of iodine absorbed per 100 g of the

oil. Mohammed (2004) reported iodine value as 105.12 g/100 g for pumpkin seed kernel oil,

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Marjana (2005) reported iodine value as 119 g/100 g for melon seed oil, Taiwo et al. (2008)

reported iodine value as 59.69 g/100 g for watermelon seed oil, Fatimah et al. (2011) reported

iodine value as 119.9-125.1 g I/100 g for winter melon seed oil, Ziyada and Elhussein (2008)

reported iodine value as 128.9 g /100 g for colocynthoide seed oil.

2.5.4 Peroxide value

Rancidity is brought about by the action of air (oxidative) or by micro organisms in oil. In

oxidative rancidity oxygen is taken by the oil or fat with the formation of peroxides. Peroxides

value is a measure of the peroxides contained in the oil. The peroxides present are determined

by titration against thiosulphate in the presence of KI using starch as indicator (Abiodun and

Adeleke, 2010). Mohammed (2004) reported peroxide value as 0.85 ± 0.46 meq peroxide /kg

for pumpkin seed kernel oil, Marjana (2005) reported peroxide value as 7.9 meq peroxide/kg

for melon seed oil, Taiwo et al. (2008) reported peroxide value as 18.7 meq peroxide/kg for

watermelon seed oil, Fatimah et al. (2011) reported peroxide value as 1.13-1.33 meq

peroxide/kg for winter melon seed oil, Ziyada and Elhussein (2008) reported peroxide value as

2.34 meq peroxide/kg for colocynthoide seed oil and for other seed oil such as cotton seed,

groundnut and sun flower as 34.1-36.02, 23.0-30.0, and 20.87 meq peroxide/kg.

2.5.5 Refractive index

It helps to determine the purity and quality of oil sample. The refractive index of a substance is

the ratio of the speed of light at a definite wavelength in vacuum to its speed in the medium.

Mohammed (2004) reported refractive index of 1.4656 ± 0.004 for pumpkin seed kernel oil,

Marjana (2005) reported refractive index (20 oC) of 1.4733 for melon seed oil, Taiwo et al.

27
(2008) reported refractive Index of 1.459 for watermelon seed oil, Fatimah et al. (2011)

reported refractive Index (25 oC) of 1.4627-1.4646 for winter melon seed oil, Ziyada and

Elhussein (2008) reported refractive index (60 oC) of 1.4589 for colocynthoide seed oil and other

seeds oil such as cotton seed, groundnut and sesame as 1.4572, 1.4550 and 1.4580.

2.5.6 Specific gravity

This is the ratio of the density of oil to the density of a standard substance at the same

temperature and pressure. For liquids and solids the standard substance is usually water.

Ziyada and Elhussein (2008) reported specific gravity (25 °C) of 0.919 g/ml for colocynthoide

seed oil, Mohammed (2004) reported specific gravity of 0.913 g/ml for pumpkin seed kernel oil,

Marjana (2005) reported specific gravity (20 oC) of 0.914 g/ml for melon seed oil, Taiwo et al.

(2008) reported specific gravity of 0.86 g/ml for watermelon seed oil, Fatimah et al. (2011)

reported specific gravity of 0.89-0.91 g/ml for winter melon seed oil. Other seeds oil such as

sesame and sunflower has specific gravity of 0.885- 0.889 g/ml (Onyeike et al., 2002).

2.5.7 Viscosity

This is the property of a fluid or semi-fluid that causes it to resist flow. Ziyada and Elhussein

(2008) reported viscosity of 38 cP for colocynthoide seed oil, and 40-50 cP for cotton seed oil.

2.5.8 Moisture content:

The determination of moisture content is one of the most important and widely used

measurements in samples that absorb and retain water. Moisture content determination look

very simple in concept, but in practice, the accurate determination is complicated by a number

of factors which vary considerably from one sample to another (Taiwo et al., 2008). Several

studies have been carried out on the chemical and functional properties of watermelon seed

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and other seeds within the cucurbitaceae family. According to Godwin et al. (2008) on research

of the proximate composition of egusi melon, showed that the moisture content of the seed 4.6

% obtained was low compared to those reported for legumes by Arkroyed and Doughty (1964)

ranging between 7.0 and 10 %. However, the value agreed closely with that reported earlier by

Ige et al. (1984) for pumpkin seeds. According to Mohammed (2004) the proximate

compositions of pumpkin whole seed and seed kernels obtained for moisture content was quite

low 5.97± 0.32 and 6.27±1.36 %, respectively and was within the range of moisture contents of

similar seeds (Kamel et al., 1982). According to Taiwo et al. (2008) the proximate composition

of watermelon, showed that the moisture content of the seed 4.91 % obtained was low

compared to those reported for legumes by Arkroyed and Doughty (1964) ranging between 7.0

and 10 %. However, the value agreed closely with that reported earlier by Ige et al. (1984) for

pumpkin seeds.

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