A PROJECT WORK SUBMITTED TO THE DEPARTMENT OF CHEMISTRY

FACULTY OF PHYSICAL SCIENCES

UNIVERSITY OF BENIN

BENIN CITY

BY

ELIKWU RUHUOMA RITA

PSC1305129

IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF

BACHELOR OF SCIENCE IN CHEMISTRY DEGREE IN PHYSICAL SCIENCES
OF THE UNIVERSITY OF BENIN NIGERIA.

AUGUST 2017

i
 CERTIFICATION

This is to certify that this project was carried out by ELIKWU RUHUOMA RITA
with Matriculation number PSC1305129 under the supervision of ** in partial
fulfillment for the award of BSc (HONS.) degree in Chemistry, University of
Benin, Benin City.

____________________ ________________
ELIKWU RITA RUHUOMA DATE
Student

_________________ ________________
PROF M.E. UKHUN DATE
Project supervisor

____________________ _________________
PROF. J. M. OKUO DATE
Head of Department

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 DEDICATION

This project is dedicated to God almighty that made this work a success and
provided everything I needed to my amazement and also to my uncle and his
wife Mr. and Mrs. Wenekanma, my Dad Mr. Godspower Elikwu, my brothers
and of course Rev. P.N. Utomi.

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 ACKNOWLEDGEMENT

This research project couldnt have been possible without the genuine and
selfless assistance of those who contributed to make it a success.

My utmost gratitude goes to God my father for the strength and favor in
completing this work and also for the completion of my four years in the
University of Benin.

Sincere gratitude goes to my supervisor Prof M.E. Ukhun for being a father
and for his immense contributions and corrections.

To my HOD Prof J.M. Okuo and the entire staff of Chemistry Department I say a
very big thank you.

To my spiritual father Rev. P.N. Utomi thank you sir. I love you sir. To my
awesome friend Ozemoya, Mr Wells, Grace Agbator, Mr Clinton, Bro Excel
Eliboh, S. Cheryl, S. Endy, S. Betty, B. Kenny, B.Nathan, B. BOBO, Feji Boo, Frida,
Majestic B., James Divine. I love you all specially.

And of course my family Mr. Godspower Elikwu, Major Chidi Wenekanma

(rtd), Mrs. Grace Wenekanma, Bright Elikwu, Elvis Elikwu, Rogers Elikwu and
Charles Elikwu. Ill do good by you.

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 TABLE OF CONTENTS

CERTIFICATION ................................................................................................................... ii
DEDICATION ........................................................................................................................ iii
ACKNOWLEDGEMENT ..................................................................................................... iv
TABLE OF CONTENTS ....................................................................................................... v
LIST OF TABLES AND FIGURES ................................................................................ viii
ABSTRACT ............................................................................................................................. ix
CHAPTER ONE ...................................................................................................................... 1
1.0 INTRODUCTION ........................................................................................................... 1
1.1 AIM OF STUDY ......................................................................................................... 3
1.2 OBJECTIVES OF STUDY ....................................................................................... 3
1.3 RELEVANCE OF STUDY ....................................................................................... 4
LITERATURE REVIEW ...................................................................................................... 5
VEGETABLE OILS ................................................................................................................ 5
FATTY ACIDS IN VEGETABLE OILS ............................................................................. 5
TYPES OF VEGETABLE OILS........................................................................................... 6
PALM OIL ................................................................................................................................ 6
COMPOSITION ...................................................................................................................... 7
PROCESSING AND USE...................................................................................................... 9
REFINING................................................................................................................................ 9
RED PALM OIL ...................................................................................................................... 9
COCONUT OIL .................................................................................................................... 12
COCONUT PALM ............................................................................................................... 12
PROCESSING....................................................................................................................... 12
COMPOSITION ................................................................................................................... 13

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GROUNDNUT OIL (Peanut oil).................................................................................... 14
HISTORY AND EXTRACTION ....................................................................................... 14
CHEMICAL AND PHYSICAL CHARACTERISTICS OF GROUNDNUT OIL .... 15
COLOUR ................................................................................................................................ 15
DENSITY AND VISCOSITY............................................................................................. 15
FATTY ACIDS...................................................................................................................... 16
IODINE VALUE...................................................... Error! Bookmark not defined.
ACETYL VALUE ................................................................................................................. 16
HEAT OF FUSION.............................................................................................................. 16
GROUNDNUT OIL USES ................................................................................................. 17
FRYING AND FOOD .......................................................................................................... 17
SOYABEAN OIL .................................................................................................................. 17
RECOVERY AND REFINING OF SOYBEAN OIL ..................................................... 18
COMPOSITION OF SOYBEAN AND SOYBEAN OIL .............................................. 19
FOOD USES AND APPLICATION OF SOYBEAN OIL ............................................ 19
IODINE VALUE................................................................................................................... 20
UV SPECTROSCOPY ......................................................................................................... 24
PRINCIPLE OF UV SPECTROSCOPY .......................................................................... 24
ULTRAVIOLET-VISIBLE SPECTROPHOTOMETER ............................................. 25
INSTRUMENTATION AND WORKING OF UV SPECTROSCOPY..................... 28
APPLICATIONS OF UV SPECTROSCOPY ................................................................. 30
ADDITIONAL APPLICATIONS ..................................................................................... 31
CHAPTER TWO.................................................................................................................. 32
MATERIALS AND METHODS ....................................................................................... 32
SOURCE OF MATERIALS ............................................................................................... 32
OIL SAMPLES ..................................................................................................................... 32
REAGENTS........................................................................................................................... 32

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APPARATUS ........................................................................................................................ 33
PREPARATION OF REAGENTS ................................................................................... 34
DETERMINATION OF IODINE VALUE OF THE OILS ......................................... 35
MATERIALS FOR UV ABSORBANCE AT 233nm .................................................. 35
ABSORBANCE .................................................................................................................... 36
CHAPTER THREE ............................................................................................................. 37
RESULTS AND DISCUSSION ......................................................................................... 37
RELATIONSHIP BETWEEN ABSORBANCE AND IODINE VALUE ................. 37
PALM OIL ............................................................................................................................. 37
COCONUT OIL .................................................................................................................... 38
GROUNDNUT OIL ............................................................................................................. 39
SOYABEAN OIL .................................................................................................................. 40
EFFECT OF TIME ON IODINE VALUE ON THE OIL SAMPLES ....................... 41
EFFECT OF TIME ON THE ABSORBANCE OF THE OIL SAMPLES................ 42
CONCLUSION...................................................................................................................... 44
RECOMMENDATIONS .................................................................................................... 45
REFERENCES...................................................................................................................... 46
APPENDIX............................................................................................................................ 50

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 LIST OF TABLES AND FIGURES

Table 2.1 33

Fig 3.1 37

Fig 3.2 38

Fig 3.3 39

Fig 3.4 40

Fig 3.5 41

Fig 3.6 42

Table I 50

Table II 50

Table III 51

Table IV 51

Table V 52

Table VI 52

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ABSTRACT

ix
 CHAPTER ONE

1.0 INTRODUCTION

At the present time, vegetable oils are the source of most of the visible fat in
the Nigerian diet. They are used as salad and cooking oils, in salad dressing,
margarine and shortening. Processing methods include extraction, refining,
hydrogenation and trans esterification. They consist of building blocks called
triglycerides resulting from the combination of one unit of glycerol and
three units of fatty acids. They are insoluble in water but quite soluble in most
organic solvents. They have lower densities than water, and may have been
observed to have consistencies at ambient temperature of solid, semi- solid or
clear liquid. They are referred to as fats when they are solid appearing at
normal room temperature, and when they are liquid at that temperature, they
are called oils.

One of the ways of obtaining the degree of carbon carbon unsaturation of

oils is the iodine value experiment. This unsaturation is in the form of double
bonds which react with iodine compounds. The iodine value or the iodine
adsorption value is defined as the number of grams of iodine required to
saturate 100 grams of fat or oil. The iodine value or number is also an index
for assessing the ability of the oil to go rancid (Amoo et al., 2004).

Animal and vegetable fats and oils are mixtures of triglycerides. A fat or oil
that is high in unsaturated triglycerides will have a high iodine value. Many
vegetable oils are rich in unsaturated triglycerides. Sunflower oil, for example,
has an iodine value of 110-143, compared with 35-48 for a typical animal fat.
Coconut oil, in contrast, is highly saturated, with an iodine value of only 6-11.
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One of the properties of unsaturated organic compounds is the reactivity of
the double bonds, especially their ability to form addition compounds with
halogens. As the addition takes place at the double bond, measurement of the
quantity absorbed is a measure of the number of double bonds present
(Ihekoronye and Ngody, 1985). Low iodine number implies the presence of
few unsaturated bonds and hence low susceptibility to oxidative rancidity
(Onyeike and Oguike, 2003). High iodine values signify or indicate high
content of poly unsaturated fatty acid in the product and its susceptibility to
rancidity.

Recent studies have showed that the higher the degree of unsaturation (i.e.
high iodine value), the greater the tendency of the fat to oxidative rancidity.
Asibuo et al. (2008) analyzed oils from twenty groundnut varieties and
indicated that the iodine values ranged from 87.77 to 98.43mg/100g. Ayo
(2016) also showed that the iodine value of the oil samples obtained from
Soya bean oil was (1.380mg KOH/g) and Groundnut oil (Kings refined oil) was
(1.795mg KOH/g).

The properties and characteristics of oil have widely been studied with the aid
of ultraviolet, visible, infrared and the X-ray regions of the electromagnetic
spectrum. In particular, studies within this region have helped in resolving
some problem in the oil chemistry and analysis studies.

UV spectroscopy is type of absorption spectroscopy in which light of ultra-

violet region (200-400 nm.) is absorbed by the molecule. Absorption of the
ultra-violet radiations results in the excitation of the electrons from the
ground state to higher energy state. The energy of the ultra-violet radiation
that are absorbed is equal to the energy difference between the ground state
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and higher energy states (deltaE = hf). UV/Vis spectroscopy is routinely used
in analytical chemistry for the quantitative determination of different
analytes, such as transition metal ions, highly conjugated organic compounds,
and biological macromolecules.

The ultraviolet spectroscopy can provide information on the quality of a fat,

its state of preservation and changes brought about in it by technological
processes.

Spectroscopic analysis is commonly carried out in solutions but solids and

gases may also be studied. It has been used by Ballabio (2006) and Haddada
(2007), where the absorption at the wavelengths specified in the method was
due to the presence of conjugated diene and triene systems. The analysis of
Ayo (2016) after conducting tests on three different vegetable oils revealed
that the UV absorbance at 233nm of the oil samples was high for palm oil
(0.236) as compared to the absorbance of groundnut oil (0.015). The lowest
was with soya bean oil (0.007) indicating the decreasing level of conjugation
in the oils.

1.1 AIM OF STUDY

To determine the relationship between Iodine value and absorbance at

233nm (UV spectroscopy) of selected oils on heating at different time
intervals.

1.2 OBJECTIVES OF STUDY

To determine the iodine value of vegetable oil

To determine the absorbance of these oils at a wavelength of 233nm.

3
 To determine the effect of time on the iodine values and absorbance of
these vegetable oils.
To show the relationship between the acid value and absorbance at
233nm of the various vegetable oils.
Plot absorbance value against iodine value for each oil sample and
calculate the correlation coefficient from the resulting plot.
Work out the regression of the plot i.e. y = mx + c.

1.3 RELEVANCE OF STUDY

The ultraviolet spectroscopy can provide information on the quality of a

fat, its state of preservation and changes brought about in it by
technological processes.
Determination of the composition of oil, Measurement of cis-trans
isomerization of Oil, Notable include: Measurement of the degree of
oxidation of oil, and determination of the degree of polymerization of oil
(Dibie, 2015).

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 LITERATURE REVIEW

VEGETABLE OILS

Most vegetable oils are obtained from beans or seeds, which generally furnish
two valuable commodities; oil and a protein rich meal. Oil extraction is
achieved by pressing and/or by solvent extraction. Oils such as palm and olive
on the other hand, are pressed out of the soft fruit (endosperm). Seeds give
oils in different proportions. Using recent figures, world average oils yields
are: soybean (18.3%); rapeseed (38.6%); sunflower (40.9%); groundnut
(40.3%); cotton seed (15.1%); coconut (62.4%); palm kernel (44.6%) etc. The
refining processes remove undesirable materials from the oils such as
phospholipids, monoacylglycerols, diacylglycerols, free acids, colours and
pigments, oxidized materals, flavor materials and trace metals but may also
remove valuable minor components which are antioxidants and vitamins such
as carotenes and tocopherols.

FATTY ACIDS IN VEGETABLE OILS

Unsaturated fatty acids present in vegetable oils are characterised by high

absorbability and antiallergic properties. Thanks to their beneficial and
diverse effects on the skin they have found wide applications in many
branches of industry, in particular in cosmetic industry and cosmetology,
pharmacy and medicine. In cosmetic industry, vegetable oils are used mainly
as the vehicle for other active ingredients, dissolved or dispersed in oil-water
type emulsions. The most often used essential unsaturated fatty acids are
those from the omega-3, omega-6 and omega-9 series.

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The most important among them are 18-carbon acids (C18):
monounsaturated (omega-9) oleic acid, biunsaturated (omega-6) linoleic
acid, including cis linoleic acid with conjugated double bonds at positions 3
and 6 (CLA, conjugated-linoleic acid), triunsaturated

(omega-3), -linoleic acid (ALA) and (omega-6), -linoleic acid

TYPES OF VEGETABLE OILS

PALM OIL

Palm oil is an edible vegetable oil derived from the mesocarp (reddish pulp) of
the fruit of the oil palms, primarily the African oil palm Elaeis guineensis, and
to a lesser extent from the American oil palm Elaeis oleifera and the maripa
palm Attalea maripa.

Palm oil is naturally reddish in color because of a high beta-carotene content.

It is not to be confused with palm kernel oil derived from the kernel of the
same fruit, or coconut oil derived from the kernel of the coconut palm (cocos
nucifera). The differences are in color (raw palm kernel oil lacks carotenoids
and is not red), and in saturated fat content: palm mesocarp oil is 41%
saturated, while palm kernel oil and coconut oil are 81% and 86% saturated
fats, respectively.

Along with coconut oil, palm oil is one of the few highly saturated vegetable
fats and is semi-solid at room temperature. Like most plant-based products,
palm oil contains very little cholesterol.

Palm oil is a common cooking ingredient in the tropical belt of Africa,

Southeast Asia and parts of Brazil. Its use in the commercial food industry in
6
other parts of the world is widespread because of its lower cost and the high
oxidative stability (saturation) of the refined product when used for frying.

The use of palm oil in food products has attracted the concern of
environmental activist groups; the high oil yield of the trees has encouraged
wider cultivation, leading to the clearing of forests in parts of Indonesia and
Malaysia in order to make space for oil-palm monoculture. This has resulted
in significant acreage losses of the natural habitat of the orangutan, of which
both species are endangered; one species in particular, the Sumatran
orangutan, has been listed as critically endangered. In 2004, an industry
group called the Roundtable on Sustainable Palm Oil (RSPO) was formed to
work with the palm oil industry to address these concerns. Additionally, in
1992, in response to concerns about deforestation, the Malaysian Government
pledged to limit the expansion of palm oil plantations by retaining a minimum
of half the nation's land as forest cover. These commitments have not been
met.

COMPOSITION

Fatty acids

Palm oil, like all fats, is composed of fatty acids, esterified with glycerol. Palm
oil has an especially high concentration of saturated fat, specifically, of the 16-
carbon saturated fatty acid palmitic acid, to which it gives its name.
Monounsaturated oleic acid is also a major constituent of palm oil. Unrefined
palm oil is a large natural source of tocotrienol, part of the vitamin E family.

7
Carotenes

When unrefined or when processed into red palm oil, it is naturally rich in
carotenes, which give it its characteristic dark red color. Like tomatoes,
carrots and many other fruits and vegetables but unlike most oils, palm oil
naturally contains the nutrients alpha-carotene, beta-carotene and lycopene.
Palm oil contains other carotenes including tocopherols and tocotrienols
(members of the vitamin E family), CoQ10, phytosterols, and glycolipids.

Tocopherols and tocotrienols (tocols)

Crude palm oil, besides being rich in pro- vitamin A, has a high content of
vitamin E, present as tocopherols and tocotrienols, of which 70% are
tocotrienols (Hashimoto et al. 1980). A new vitamin E (- tocomonoenol)
was detected and identified by Matsumoto et al. (1995) and further examined
during physical refining (Puah et al. 2007).

Sterols, Squalene and other hydrocarbons

Another major component group of the unsaponifiable fraction of palm oil are
the phytosterols. The common phytosterols found in palm oil products are
sitosterol, stigmasterol campesterol and cholesterol. Crude palm oil contains
210620 ppm of phytosterols. Fractionation and refining changes the content
and composition of the phytosterols in the oil and its fractions. Again, palm
fatty acid distillate is a good source of phytosterols, having 150020000 ppm
with an average of 6500 ppm (Abdul Gapor et al. 1988). In the extraction of
palm tocotrienols, the process involves a purification step that results in a
concentrate high in the phytosterols

8
PROCESSING AND USE

Many processed foods either contain palm oil or various ingredients derived
from it.

REFINING

After milling, various palm oil products are made using refining processes.
First is fractionation, with crystallization and separation processes to obtain
solid (stearin), and liquid (olein) fractions. Then melting and degumming
removes impurities. Then the oil is filtered and bleached. Physical refining
[clarification needed] removes smells and coloration to produce "refined,
bleached and deodorized palm oil" (RBDPO) and free sheer fatty acids, which
are used in the manufacture of soaps, washing powder and other products.
RBDPO is the basic palm oil product sold on the world's commodity markets.
Many companies fractionate it further to produce palm olein for cooking oil,
or process it into other products.

RED PALM OIL

Since the mid-1990s, red palm oil has been cold-pressed and bottled for use as
cooking oil, and blended into mayonnaise and salad oil.

The highly saturated nature of palm oil renders it solid at room temperature
in temperate regions, making it a cheap substitute for butter or trans fats in
uses where solid fat is desirable, such as the making of pastry dough and
baked goods. A recent rise in the use of palm oil in the food industry has partly
come from changed labelling requirements that have caused a switch away
from using trans fats. Palm oil has been found to be a reasonable replacement

9
for trans fats; however, a small study conducted in 2009 found that palm oil
may not be a good substitute for trans fats for individuals with already-
elevated LDL levels. The USDA agricultural research service states that palm
oil is not a healthy substitute for trans fats.

Biomass and Bioenergy

Palm oil is used to produce both methyl ester and hydrodeoxygenated

biodiesel. Palm oil methyl ester is created through a process called
transesterification. Palm oil biodiesel is often blended with other fuels to
create palm oil biodiesel blends. Palm oil biodiesel meets the European EN
14214 standard for biodiesels. Hydrodeoxygenated biodiesel is produced by
direct hydrogenolysis of the fat into alkanes and propane. The world's largest
palm oil biodiesel plant is the Finnish-operated Neste Oil biodiesel plant in
Singapore, which opened in 2011 and produces hydrodeoxygenated NEXBTL
biodiesel.

The organic waste matter that is produced when processing oil palm,
including oil palm shells and oil palm fruit bunches, can also be used to
produce energy. This waste material can be converted into pellets that can be
used as a biofuel. Additionally, palm oil that has been used to fry foods can be
converted into methyl esters for biodiesel. The used cooking oil is chemically
treated to create a biodiesel similar to petroleum diesel.

Cooking/frying oil

Palm olein is much utilised as cooking oil in homes and in industrial outlets.
Palm oil and its fractions are accepted as frying oils for food products such as
snack chips, crackers, cookies, pastries, doughnuts, fries and instant noodles.

10
A comprehensive review of palm oil products in frying applications has been
documented by Berger (2007). Frying, being a thermal process carried out in
air, generally results in a rapid deterioration of the oil. The
oxidative stability of palm oil, olein and stearin is a major advantage of these
oils. Palm olein has the longest induction period: 44 hours at 100 C. Blending
less stable vegetable oils with palm olein improves their stability. The
improvements are seen in the reduced levels of primary and secondary
oxidation products, fatty acids, volatiles and polymers. The cloud points of
palm olein with unsaturated oil blends are also improved.

The free fatty acid content is one of the parameters used for evaluating the
quality of frying oils. During frying, there is a lower formation of free acids
when palm olein is used or blended with other vegetable oils. Besides this, the
polymer content is lower and so less change in viscosity is observed. Most
polyunsaturated oils have to be hydrogenated for use as frying oils, to reduce
high polymer formation and consequent viscosity increase during frying. This
leads to the undesirable presence of trans acids in the frying oil.

Margarines
It is a legal requirement that margarine contain at least 80% fat. Many
products are now available with lower levels of fat and these should be
designated as spreads rather than as margarines. However, in this account of
margarines the term margarine is used to include the reduced- fat spreads.
Margarine is a product containing 80% fat blended with water, and containing
vitamins and other ingredients. It was initially developed to replace dairy
butter and now appears in a variety of types, which include regular, whipped,
soft-tub, liquid, diet, low- calorie, bakery, speciality and so on. Todays

11
margarines incorporate nutritional as well as functional properties and cater
for the requirements of different consumers. The properties of margarines
depend on the characteristics of the oil, which is the major ingredient of the
product. The solid fat content of the oil at different temperatures is an
indicator of the crystallization properties of the finished product.

Palm oil and its fractions are suitable for margarine production, as shown by
Nor Aini and Mohd Suria (2000).

COCONUT OIL

COCONUT PALM

Coconut palm is productively grown within 20 north and south of the

equator, especially along coastal areas (Gunstone and Harwood 2007). There
are two types of coconut palm: the tall and the dwarf type. The tall coconut
palm gives higher oil yields than the dwarf type because the tough copra
obtained from the latter makes it unsuitable for commercial purposes. It is
planted mainly as an ornamental. The more common tall variety reaches a
height of over 30 m and has a lifespan of more than 50 years (Canapi et al.
2005). Typically, fresh coconut kernel contains (% wt.) moisture (50), oil (34),
ash (2.2), fiber (3.0), protein (3.5) and carbohydrate (7.3) (Canapi et al. 2005).

PROCESSING

The first step in coconut oil extraction is dehulling; that is, cracking the shell
to take out the meat or kernel. The kernel contains about 50% moisture and it
has to be dried to a moisture content of 68% before oil extraction. This can
be achieved by drying the kernel under the sun, with direct heat or through

12
the use of hot air. The dried kernel is known as copra and has an oil content of
64%. Traditionally, coconut oil is extracted from the copra by crushing in an
expeller, followed by solvent extraction to recover the residual oil from the
cake (Canapi et al. 2005). The crude oil is then refined by physical or chemical
refining to remove impurities, making it suitable for human consumption and
prolonging its shelf-life. In physical refining, the crude oil is firstly pre-treated
with 0.050.1% aqueous phosphoric acid (85%) and heated to 8090 C for
2030 min, then bleached with a mixture of bleaching clay/activated carbon
(10:1 ratio) at 9095 C for 2030 min and finally by deodorization at 240 C
for 11.5 h, with steam injected at the bottom of the column (Canapi
et al. 2005). Phospholipids are removed during the pre-treatment. Color
bodies, metal ions, phosphoric acid and other adsorbable impurities are
removed during bleaching, along with free fatty acids (FFA) and volatile
components. In chemical refining the FFA in the oil is neutralized with sodium
hydroxide and removed in a water stream at an early stage.

COMPOSITION

The major component of crude oils is triacylglycerol (TAG) (about 95%),

while the minor components comprise free acids, monoacylglycerols,
diacylglycerols, phospholipids, free and/or acylated sterols, tocols and
hydrocarbons such as alkanes, squalene and carotenes (Gunstone 2006).
About 0.5% of crude coconut oil is not saponified by caustic treatment.
This unsaponifiable matter consists mainly of tocols, sterols, squalene, color
pigments, carbohydrates and odour compounds (lactones) (Canapi et al.
2005). Most of the unsaponifiables are removed during the refining, bleaching
and deodorising of crude coconut oil. The crude oil also contains protein,

13
crude fibre and trace amounts of metals such as iron, copper
and lead.

GROUNDNUT OIL (Peanut oil)

HISTORY AND EXTRACTION

Groundnut oil is expressed from the seed of Arachis hypogaea L., commonly
known as groundnut, peanut, or earth nut because the seeds develop
underground. Groundnuts are produced on a significant basis in more than 30
countries, with worldwide production figures estimated to be in excess of 30
million metric tons.

The world production of groundnuts is about 7% of the world production of

oilseeds, but because of the limited extraction of groundnuts, the annual
production of groundnut oil is only about 4% of the world vegetable oil total
Uses in various countries differ greatly, but overall more than 50% of all
groundnuts produced are crushed for oil. Whereas in India 7580% of the
groundnut crop is crushed for oil due to high demand, in contrast in the US
only 1012% of the groundnuts produced are crushed. This low percentage is
indicative of the economic importance of the nuts themselves as a food crop in
the US. Due to the high content of digestible protein and unsaturated oil and
the exceptional roasted nutty flavor, groundnuts have substantial value as a
nutritious and flavorful food commodity. More than one third of the
groundnuts produced are used as food in the form of intact nuts on a
worldwide basis. In the US, a high percentage of the limited numbers of
groundnuts used for oil extraction have been separated from edible stocks
because of the potential for aflatoxin contamination. After minimal oil

14
extraction, the pressed cake, which is low in oil and high in protein, may be
used for animal feed if aflatoxin is kept below acceptable levels.

CHEMICAL AND PHYSICAL CHARACTERISTICS OF GROUNDNUT OIL

COLOUR

As peanuts mature, oil color becomes lighter as -carotene and lutein, which
are responsible for the yellow color, become more diluted (Pattee and Purcell
1967). Although oil color may be used to assess maturity, other methods are
preferred, because many factors such as curing temperature and duration
influence oil color (Sanders et al. 1982). Color measurement is frequently
done by visual comparison under a standard light source or Gardner color,
which has a scale between 1 and 18.

DENSITY AND VISCOSITY

Viscosity and density are important physical parameters central to the quality
of vegetable oils. These properties, as summarized in Table 8.8, were surveyed
as a function of temperature (5C to 100C) for oils from nine common
cultivars of peanut to determine the potential for variation (Davis et al. 2008).
Increasing content of oleic acid, decreasing content of linoleic acid, and
decreasing content of palmitic acid were each associated with decreased
density and increased viscosity among the oils. High-oleic oils had both the
lowest densities and highest viscosities, with viscosity differences being most
apparent at cooler temperatures. Non-linearity of hydrocarbon chains due to
unsaturation was considered to affect oil density and viscosity.

15
FATTY ACIDS

Groundnut oil contains a high proportion of unsaturated fatty acids, in

particular oleic (18:1), linoleic (18:2), and 11-eicosenoic (20:1). The saturated
fatty acids in groundnut oil are palmitic (16:0), stearic (18:0), arachidic (20:0),
behenic (22:0), lignoceric (24:0), and hexacosanoic (26:0). Palmitic is the only
saturated fatty acid that exceeds 10%. The very long-chain fatty acids (those
above 22 carbons) are usually found at or about 2% each. These fatty acids
have been associated with widely varying effects, such as the metabolism
of the dietary fatty acids and the physical properties of the oils themselves
(Dean and Sanders 2009). The oxidative stability of groundnut oil is highly
correlated with the ratio of oleic acid to linoleic acid (Fore et al. 1953). This
ratio generally increases with seed maturity and oil stability increases
simultaneously.

ACETYL VALUE

The acetyl value is the number of milligrams of KOH (potassium hydroxide)

required to neutralize the acetic acid produced by the hydrolysis of 1 g of
acetylated fat and is a measure of free hydroxyl groups present in the oil. The
acetyl number of peanut oil (8.59.5) is lower than other vegetable oils, but
higher than coconut oil, palm oil, and the animal fats and oils.

HEAT OF FUSION

The heat of fusion, or latent heat, is the quantity of heat required to change 1 g
of solid to a liquid with no temperature change. This latent heat increases with
increasing molecular weight. The heat of fusion of peanut oil is 21.7cal/g.

16
GROUNDNUT OIL USES

FRYING AND FOOD

Throughout the world, frying and cooking constitute by far the greatest use of
peanut oil. It is especially suitable for deep-fat frying due to its high smoke
point of 229 C (Woodruff 1983). This high temperature allows food to cook
quickly with a crisp coating and little oil absorption. Off flavor and odor
development are very limited during frying with groundnut oil. However,
degradation of triacylglycerols occurring during frying results in an increase
of free fatty acids (FFA) and a decrease in smoke point. In a comparison of
various frying oils on consumer acceptability of salted, fried peanuts, peanuts
prepared from refined peanut oil were better accepted than those prepared
with other vegetable oils such as sunflower, corn, soybean, and olive oils
(Ryan et al. 2008).

SOYABEAN OIL

Soyabean is the dominant oilseed produced in the world, because of its

favorable agronomic characteristics, its high-quality protein, and its valuable
edible oil. It contributes about 47% of all oilseeds produced worldwide in
2008/09. The US ranks first in soyabean oil production (8.5 million tonnes),
followed by China, Argentina, Brazil, EU-27, and India (7.3, 6.1, 6.0, 2.3, and
1.3 million tonnes, respectively). The production of soyabean and soyabean oil
is driven by the need for soy protein meal, which is used extensively in
commercial feeds for poultry, swine, and cattle. Soyabean oil accounted for
about 80% of total edible oil consumption in the US (USDA-NASS) in 2008
because of its availability and its many desirable characteristics, including

17
compositional and functional properties. Soyabean oil was the predominant
vegetable oil produced in the world until 2003/04, but is now surpassed by
palm oil.

RECOVERY AND REFINING OF SOYBEAN OIL

Oil extraction

The two common processes for soyabean oil extraction are solvent extraction
and mechanical pressing, but in the US less than 1% of soyabeans is processed
by mechanical means. Solvent extraction with hexane is the standard practice
in todays modern processing facilities, and its use has been reviewed by
Johnson (2008). There are three major steps in solvent extraction: seed
preparation, oil extraction, and desolventizing of the oil and meal.
Conventional seed preparation includes drying, cleaning, cracking, optional
dehulling or decortication, conditioning, and flaking of the seeds. The option
of expanding after flaking is used to improve oil extraction, percolation, and
solvent drainage, and is accompanied by a doubling of the throughput. In
another variation in seed preparation (hot dehulling), hulls are removed from
the split seeds by alternate slow and rapid heating before cracking and
flaking. Hot de-hulling is more energy efficient than conventional dehulling.
The Alcon process (Penk 1986) is a flake-heating treatment aimed to improve
the degumming efficiency of the crude soyabean oil. A very low-level PL in
degummed oil can be achieved and the oil can then be physically refined.
However, in the US the majority of soybean oil is chemically refined. Solvent
(hexane) extraction of soybeans is a diffusion process achieved by immersing
the solid in solvent or by percolating solvent through a bed of solids. Rotary
(deep-bed), horizontal belt, and continuous loop extractors are used for
18
soybeans (Woerfel 1995). Solvent is recovered from the mixture of solvent
and extracted oil (miscella) by a double-effect evaporator and steam stripping
and from flake by a desolventizer-toaster, and is recycled

COMPOSITION OF SOYBEAN AND SOYBEAN OIL

Seed composition

Mature soybeans are oval shaped and their sizes are variety dependent. The
seed consists of three major parts: seed coat or hull, cotyledon, and germ or
hypocotyls.

Oil composition

Crude oil recovered by solvent extraction or mechanical pressing contains

various classes of lipids. It consists primarily of neutral lipids, which include
tri-, di-, and mono-acylglycerols, free fatty acids, and polar lipids such as
phospholipids. It also contains a minor amount of unsaponifiable matter that
includes phytosterols, tocopherols,and hydrocarbons such as squalene. Trace
metals are found in soybean oil in ppm concentration. When the oil is refined,
concentrations of minor constituents are reduced.

FOOD USES AND APPLICATION OF SOYBEAN OIL

According to the 2009 Soya and Oilseed Bluebook, 83% of all soybean oil
produced in the US in 2006/07 was used in foods and 17% was used in non-
food applications. Out of the total oil used in food, 35% was used for
shortening production, 6% for margarine, and 58% as cooking and salad oil.
Ten years earlier these values were 97% of the total for food use, and, out of
the total, 37%, 13%, and 49% were used in shortening, margarine, and
19
cooking and salad oil, respectively. Warner (2008) also reported similar
soybean oil utilization data for 2005; that is, soybean oil used as shortening,
margarine, and cooking/salad oil at 45%, 5%, and 48%, respectively. The
general decrease in use in margarine and increase in use in cooking and salad
oil may reflect the desire to reduce the intake of trans acids as well as the use
of reduced fat spreads.

Food

Soybean oil is mostly used for frying and baking. It is also used as a condiment
for salads.

Drying oils

Soybean oil is one of many drying oils, which means that it will slowly harden
(due to free-radical based polymerization) upon exposure to air, forming a
flexible, transparent, and waterproof solid. Because of this property, it is used
in some printing ink and oil paint formulations. However, other oils (such as
linseed oil) may be superior for some drying oil applications.

Fixative for insect repellents

While soybean oil has no direct insect repellent activity, it is used as a fixative
to extend the short duration of action of essential oils such as geranium oil in
several commercial products.

IODINE VALUE

Iodine is a chemical element with symbol I and atomic number 53. The name
is from Greek ioeids, meaning violet or purple, referring to the color
of iodine vapor. Iodine is found on Earth mainly as the highly water-soluble
20
iodide ion I, which is concentrated in oceans and brine pools. Like the other
halogens, free iodine occurs mainly as a diatomic molecule I 2, and then only
momentarily after being oxidized from iodide by an oxidant like free oxygen.
In the universe and on Earth, iodine's high atomic number makes it a
relatively rare element. Present in sea water, it is the heaviest essential
element used widely by life in biological functions (only tungsten, employed in
enzymes by a few species of bacteria, is heavier). Iodine is rare in many soils,
has low abundance generally as a crust-element, and is leached by rainwater,
leading to many deficiency problems in land animals and inland human
populations. Iodine deficiency affects about two billion people and is the
leading preventable cause of intellectual disabilities.

It is used in analytical chemistry to measure the amount of unsaturation of

oils and fats. Animal and vegetable oils and fats known chemically as
triglycerides have chains of carbon atoms that can bond with hydrogen. When
the carbon atoms in these chains are bonded to the maximum possible
number of hydrogen atoms, the triglyceride is said to be saturated, but when
there are one or more double bonds between carbon atoms, there is less
hydrogen in the molecule, and the fat is said to be unsaturated. Triglycerides
with one double bond are known as monounsaturates and those with more
than one double bond are known as polyunsaturates. Iodine can combine with
fats that have carbon double bonds and, therefore, the number of such bonds
can be deduced from the amount of iodine with which they will combine.

Hydrogen and the halogen elements fluorine, chlorine, bromine and iodine
resemble one another in that they are one electron short of a stable
configuration and can form stable compounds by sharing an electron pair with

21
another atom. In a carbon-hydrogen bond, the hydrogens single electron and
one electron from the carbon are shared to form a single covalent bond.
Where there is a carbon double bond in an unsaturated fat, each of the carbon
atoms can instead form a bond with a halogen.

The more carbon double bonds an unsaturated fat has, the more halogen
atoms with which it can combine. It is therefore possible to determine the
degree of unsaturation of a fat by allowing it to combine with a halogen. A
simple test for unsaturated fats is to mix the fat with a solution of bromine in
carbon tetrachloride; if the fat is unsaturated, the brown or yellow color of the
bromine disappears as it combines with the fat. For determining the degree of
unsaturation, however, iodine is normally used, as it is easy to measure
precisely how much iodine has been used up.

To obtain the iodine value also known as the iodine number or iodine
adsorption value of oil, a known quantity of the oil is dissolved in a suitable
solvent, such as chloroform, and mixed with an excess of iodine in the form of
iodine monochloride (ICl), as this reacts more easily. Where there is a carbon
double bond, one carbon atom will form a single bond with the chlorine in the
iodine monochloride and the other with the iodine. When the reaction is
complete, potassium iodide is added to the remaining iodine monochloride to
release the iodine:

ICl + 2KI KCl + I2.

The remaining iodine is reacted with a starch to form a dark blue compound.
Sodium thiosulfate solution at a known concentration is then slowly added.
The iodine reacts with this to form colorless I- ions.

22
One of the commonly used methods for the determination of iodine value is
the wijs method. The lipid analysed is weighed and dissolved in wijs reagent.

R-CH=CH-R + ICl R-CHI-ClCH-R + ICl

The amount of iodine chloride that did not react with the molecules is added
to potassium iodide. The reaction produces iodine.

ICl + 2KI KCl + KI + I2

The iodine liberated is then titrated with sodium thiosulphate in the presence
of starch to determine the concentration of iodine released.

I2 + Starch + 2Na2S4O3 (Blue) 2NaI + Starch + Na2S4O6 (Colourless)

Once all the iodine has reacted, the solution will become colorless. At this
point, the amount of sodium thiosulfate used can be determined, and from
this, the amount of iodine that was present. When this amount is known, the
amount of iodine that reacted with the fat can be calculated, giving the iodine
value, which is expressed as grams of iodine used per 100 grams of fat.

Animal and vegetable fats and oils are mixtures of triglycerides. A fat or oil
that is high in unsaturated triglycerides will have a high iodine value. Many
vegetable oils are rich in unsaturated triglycerides. Sunflower oil, for example,
has an iodine value of 110-143, compared with 35-48 for a typical animal fat.
Coconut oil, in contrast, is highly saturated, with an iodine value of only 6-11.

There are two other numbers that may be associated with fats and oils. The
saponification number is an indication of the average molecular weight of a fat
and is determined by breaking it down into glycerol and a fatty acid salt by
treatment with a strong alkali. The acid number indicates how much free fatty

23
acid a fat contains and is estimated from the amount of alkali required to
neutralize it.

UV SPECTROSCOPY

Ultravioletvisible spectroscopy or ultraviolet-visible spectrophotometry (UV-

Vis or UV/Vis) refers to absorption spectroscopy or reflectance spectroscopy
in the ultraviolet-visible spectral region. This means it uses light in the visible
and adjacent ranges. The absorption or reflectance in the visible range directly
affects the perceived color of the chemicals involved. In this region of the
electromagnetic spectrum, atoms and molecules undergo electronic
transitions (Skoog et al, 2007). Absorption spectroscopy is complementary to
fluorescence spectroscopy, in that fluorescence deals with transitions from
the excited state to the ground state, while absorption measures transitions
from the ground state to the excited state (Misra et al, 2002). Generally, the
most favoured transition is from the highest occupied molecular orbital
(HOMO) to lowest unoccupied molecular orbital (LUMO). For most of the
molecules, the lowest energy occupied molecular orbitals are s orbital, which
correspond to sigma bonds. The p orbitals are at somewhat higher energy
levels, the orbitals (non-bonding orbitals) with unshared paired of electrons
lie at higher energy levels (Soovali et al, 2006).

PRINCIPLE OF UV SPECTROSCOPY

UV spectroscopy obeys the Beer-Lambert law, which states that: when a beam
of monochromatic light is passed through a solution of an absorbing substance,
the rate of decrease of intensity of radiation with thickness of the absorbing

24
solution is proportional to the incident radiation as well as the concentration of
the solution (Metha, 2012).

The expression of Beer-Lambert law is given below as;

A = log (I0/I) = Ecl

Where, A = absorbance

I0 = intensity of light incident upon sample cell

I = intensity of light leaving sample cell

C = molar concentration of solute

L = length of sample cell (cm.)

E = molar absorptivity

From the Beer-Lambert law it is clear that greater the number of molecules
capable of absorbing light of a given wavelength, the greater the extent of light
absorption. This is the basic principle of UV spectroscopy.

ULTRAVIOLET-VISIBLE SPECTROPHOTOMETER

The instrument used in ultraviolet-visible spectroscopy is called a UV/Vis

spectrophotometer. It measures the intensity of light passing through a
sample (I), and compares it to the intensity of light before it passes through
the sample (Io). The ratio (I/Io) is called the transmittance, and is usually
expressed as a percentage (%T). The absorbance A is based on the
transmittance:

A = log (T/ 100

25
The UV-visible spectrophotometer can also be configured to measure
reflectance. In this case, the spectrophotometer measures the intensity of light
reflected from a sample (I)and compares it to the intensity of light reflected
from a reference material (Io) (such as a white tile). The ratio (I/Io) is called
the reflectance, and is usually expressed as a percentage (%R).

The basic parts of a spectrophotometer are a light source, a holder for the
sample, a diffraction grating in a monochromator or a prism to separate the
different wavelengths of light, and a detector. The radiation source is often a
Tungsten filament (300-2500 nm), a deuterium arc lamp, which is continuous
over the ultraviolet region (190-400 nm), Xenon arc lamp, which is
continuous from 160-2,000 nm; or more recently, light emitting diodes (LED)
(Skoog et al, 2007) for the visible wavelengths. The detector is typically a
photomultiplier tube, a photodiode, a photodiode array or a charge-coupled
device (CCD). Single photodiode detectors and photomultiplier tubes are used
with scanning monochromators, which filter the light so that only light of a
single wavelength reaches the detector at one time. The scanning
monochromator moves the diffraction grating to "step-through" each
wavelength so that its intensity may be measured as a function of wavelength.
Fixed monochromators are used with CCDs and photodiode arrays. As both of
these devices consist of many detectors grouped into one or two dimensional
arrays, they are able to collect light of different wavelengths on different
pixels or groups of pixels simultaneously.

In a double-beam instrument, the light is split into two beams before it

reaches the sample. One beam is used as the reference; the other beam passes
through the sample. The reference beam intensity is taken as 100%

26
Transmission (or 0 Absorbance), and the measurement displayed is the ratio
of the two beam intensities. Some double-beam instruments have two
detectors (photodiodes), and the sample and reference beam are measured at
the same time. In other instruments, the two beams pass through a beam
chopper, which blocks one beam at a time. The detector alternates between
measuring the sample beam and the reference beam in synchronism with the
chopper. There may also be one or more dark intervals in the chopper cycle.
In this case, the measured beam intensities may be corrected by subtracting
the intensity measured in the dark interval before the ratio is taken.

Samples for UV/Vis spectrophotometry are most often liquids, although the
absorbance of gases and even of solids can also be measured. Samples are
typically placed in a transparent cell, known as a cuvette. Cuvettes are
typically rectangular in shape, commonly with an internal width of 1 cm. (This
width becomes the path length L in the Beer-Lambert law). Test tubes can also
be used as cuvettes in some instruments. The type of sample container used
must allow radiation to pass over the spectral region of interest. The most
widely applicable cuvettes are made of high quality fused silica or quartz glass
because these are transparent throughout the UV, visible and near infrared
regions. Glass and plastic cuvettes are also common, although glass and most
plastics absorb in the UV, which limits their usefulness to visible wavelengths.
Specialized instruments have also been made. These include attaching
spectrophotometers to telescopes to measure the spectra of astronomical
features. UV-visible microspectrophotometers consist of a UV-visible
microscope integrated with a UV-visible spectrophotometer.

27
A complete spectrum of the absorption at all wavelengths of interest can often
be produced directly by a more sophisticated spectrophotometer. In simpler
instruments the absorption is determined one wavelength at a time and then
compiled into a spectrum by the operator. By removing the concentration
dependence, the extinction coefficient () can be determined as a function of
wavelength.

The iodine value is a measure of the degree of unsaturation in oils, as

determined by the uptake of appropriate halogen compounds. Because
melting point and oxidative stability are related to the degree of unsaturation,
IV provides an estimation of these quality factors. The greater the iodine
value, the more the unsaturation and the higher the susceptibility to
oxidation. Peanut oil (IV 82107) is more saturated than corn (IV 103128),
cottonseed (IV 99113), or linseed (IV 155205) oils; however, it is
considerably less saturated than coconut (IV 7.710.5), palm (IV 4454), or
butter (IV 2542) oils (Pattee 2005).

INSTRUMENTATION AND WORKING OF UV SPECTROSCOPY

Instrumentation and working of the UV spectrometers can be studied

simultaneously. Most of the modern UV spectrometers consist of the following
parts (Mark et al, 2007):

Light source- Tungsten filament lamps and Hydrogen-Deuterium lamps are

most widely used and suitable light source as they cover the whole UV region.
Tungsten filament lamps are rich in red radiations; more specifically they emit
the radiations of 375 nm, while the intensity of Hydrogen-Deuterium lamps
falls below 375 nm.

28
Monochromator- Monochromators generally composed of prisms and slits.
The most of the spectrophotometers are double beam spectrophotometers.
The radiation emitted from the primary source is dispersed with the help of
rotating prisms. The various wavelengths of the light source which are
separated by the prism are then selected by the slits such the rotation of the
prism results in a series of continuously increasing wavelength to pass
through the slits for recording purpose. The beam selected by the slit is
monochromatic and further divided into two beams with the help of another
prism.

Sample and reference cells- One of the two divided beams is passed through
the sample solution and second beam is pass through the reference solution.
Both sample and reference solution are contained in the cells. These cells are
made of either silica or quartz. Glass can't be used for the cells as it also
absorbs light in the UV region.

Detector- Generally two photocells serve the purpose of detector in UV

spectroscopy. One of the photocell receives the beam from sample cell and
second detector receives the beam from the reference. The intensity of the
radiation from the reference cell is stronger than the beam of sample cell. This
results in the generation of pulsating or alternating currents in the photocells.

Amplifier- The alternating current generated in the photocells is transferred

to the amplifier. The amplifier is coupled to a small servometer. Generally
current generated in the photocells is of very low intensity, the main purpose
of amplifier is to amplify the signals many times so we can get clear and
recordable signals.

29
Recording devices- Most of the time amplifier is coupled to a pen recorder
which is connected to the computer. Computer stores all the data generated
and produces the spectrum of the desired compound.

APPLICATIONS OF UV SPECTROSCOPY

Detection of functional groups

UV spectroscopy is used to detect the presence or absence of chromophore in
the compound. This is technique is not useful for the detection of
chromophore in complex compounds. The absence of a band at a particular
band can be seen as an evidence for the absence of a particular group. If the
spectrum of a compound comes out to be transparent above 200 nm than it
confirms the absence of:
a) Conjugation. b) A carbonyl group. c) Benzene or aromatic compounds. d)
Bromo or iodo atoms.

Detection of extent of conjugation

The extent of conjugation in the polyenes can be detected with the help of UV
spectroscopy. With the increase in double bonds the absorption shifts towards
the longer wavelength. If the double bond is increased by 8 in the polyenes
then that polyene appears visible to the human eye as the absorption comes in
the visible region.

Identification of an unknown compound

An unknown compound can be identified with the help of UV spectroscopy.
The spectrum of unknown compound is compared with the spectrum of a
reference compound and if both the spectrums coincide then it confirms the
identification of the unknown substance.
30
Determination of configurations of geometrical isomers
It is observed that cis-alkenes absorb at different wavelength than the trans-
alkenes. The two isomers can be distinguished with each other when one of
the isomers has non-coplanar structure due to steric hindrances. The cis-
isomer suffers distortion and absorbs at lower wavelength as compared to
trans-isomer.

Determination of the purity of a substance

Purity of a substance can also be determined with the help of UV
spectroscopy. The absorption of the sample solution is compared with the
absorption of the reference solution. The intensity of the absorption can be
used for the relative calculation of the purity of the sample substance.

ADDITIONAL APPLICATIONS

UV-Vis spectroscopy is also used in the semiconductor industry to

measure the thickness and optical properties of thin films on a wafer.

31
 CHAPTER TWO

MATERIALS AND METHODS

SOURCE OF MATERIALS

The samples and reagents used during the course of this experiment were
obtained from supermarkets and chemical laboratories in Benin City.

This experiment was carried out at the Department of Chemistry University of

Benin, Benin City.

OIL SAMPLES

I. Coconut oil
II. Palm kernel oil (PKO)
III. Soybean oil
IV. Groundnut oil.

The various oils were kept in bulk inside a transparent bottle in the
laboratory.

REAGENTS

I. Carbon tetrachloride
II. Iodine monochloride (Wijs reagent)
III. 1% Starch indicator
IV. 0.1N Standard sodium thiosulphate
V. 10% Potassium iodide
VI. Distilled water

32
APPARATUS

Table 2.1

ITEM FUNCTION
Conical flask For holding reagents and also a
used during titration.
Beaker For holding reagents and also a
used during titration.
Measuring cylinder For taking volumetric
measurements of reagents during
the experimental procedures.
Burette For the transfer of reagents from
one vessel to another in the
determination of the iodine
value. It is attached to the retort
stand during titration.
Drop pipette For the transfer of reagents from
one vessel to another in the
determination of the iodine
value.
Funnel Used in transferring solution
from one medium to another.
Electronic weighing balance For taking the mass in grams of
the oil used for the experiment.
Stirrers For agitating the reagents and
allowing for even distribution

33
 throughout the medium.
Water bath To cool off or reduce the
temperature of high temperature
substances.
Retort stand For holding the pipette in place
during titration.
UV Spectrophotometer For the determination of the
absorbance of the oils.
Weighing balance For obtaining the weight of the
samples and reagents during the
experiment.

PREPARATION OF REAGENTS

Starch indicator

1% Starch indicator was prepared by weighing 1g of the soluble starch and

dissolved in little quantity of distilled water to form a paste. 100ml of boiling
water was added and stirred to give 1% of the starch indicator.

Potassium iodide

10% Potassium iodide was prepared by weighing 10g Potassium iodide salt in
100ml of distilled water. 0.1N of sodium thiosulphate solution was made by
dissolving 24.9g of sodium thiosulphate crystals in 1L of distilled water and
stirred.

Wijs solution

34
1500ml of acetic acid and 600ml of carbon tetrachloride was measured using
a weighing balance. The respective amounts of the acetic acid and carbon
tetrachloride was mixed together and about 20ml of the resulting solution
was used to dissolve the 20g of iodine trichloride and further all the solutions
were mixed together.

DETERMINATION OF IODINE VALUE OF THE OILS

The iodine value test was conducted on four oils namely; groundnut oil,
soyabean oil, palm oil and coconut oil. 0.422g of palm oil, 0.433g of coconut
oil, 0.436g of groundnut oil and 0.412g of soyabean were measured into
different beakers and an extra beaker was used for the blank.
10ml of carbon tetrachloride (CCl4) was added to each of the samples and the
blank and then 25ml of wijs solution was also added to the mixture and
incubated in a dark cupboard for one hour. After the one hour duration the
samples was brought out and poured into the burette on the retort stand. To
each of the samples in the beaker 10ml of carbon tetrachloride, 10ml of
potassium iodide, 150ml of water and 2ml starch was added. The resulting
solution was titrated against sodium thiosulphate. At first, a pinkish colour
was observed but turned colorless as the titration continued. This procedure
was carried out on the respective oil samples were carried out eight times
with a two day interval.

MATERIALS FOR UV ABSORBANCE AT 233nm

I. U.V. absorption spectrophotometer

II. Quartz cuvette
III. 100 ml. volumetric flask

35
 IV. Hexane solvent for spectrophotometer analysis
V. Analytical balance

ABSORBANCE

0.1g of each of the samples was weighed and 10ml of hexane was added to
each of these samples and poured into four different test tubes representing
the four samples.
Hexane as a solvent was used as the blank for the absorbance and the
absorbance was taken using the UV spectrophotometer at a point of 233nm.

36
 CHAPTER THREE

RESULTS AND DISCUSSION

RELATIONSHIP BETWEEN ABSORBANCE AND IODINE VALUE

PALM OIL

0.8

0.7

0.6 y = 0.0576x + 0.0429

Absorbance(233nm)

R = 0.6384
0.5

0.4

0.3

0.2

0.1

0
0 10 20 30 40 50 60
Iodine value(mgKOH/g)

Fig 3.1 a plot of Absorbance (233nm) against Iodine value for palm oil

From the above Microsoft Excel plot (fig 3.1) the r-value obtained was
calculated to be -0.7487 and this indicates a negative correlation coefficient.
This is because as the absorbance increased, the iodine value decreased hence
an inversely proportional relationship between both variables. Since the
closer the correlation coefficient to +1 or -1 the stronger the relationship
between the variables, with an r-value of -0.7487, it can be deduced that the

37
absorbance and the iodine value have a negative moderately strong
relationship.

The R2 value as seen in the plot above was calculated to be 0.6384; and this
means or signifies that the iodine value of the palm oil can predict its
corresponding absorbance with a 63.84% accuracy and vice versa.

COCONUT OIL

0.09
0.08
0.07 y = 0.0045x + 0.0374
Absorbance(233nm)

R = 0.9339
0.06
0.05
0.04
0.03
0.02
0.01
0
0 2 4 6 8 10 12
Iodine value(mgKOH/g)

Fig 3.2 a plot of Absorbance (233nm) against Iodine value for coconut oil

From the above Microsoft Excel plot (fig 3.2) the calculated r-value was -
0.9841 and this indicates a highly negative correlation coefficient. This is
because as the absorbance increased, the iodine value decreased hence an
inversely proportional relationship between both variables. Since the closer
the correlation coefficient to +1 or -1 the stronger the relationship between
the variables, with an r-value of -0.9841, it can be deduced that the

38
absorbance and the iodine value have an almost perfect linear relationship
since the r-value is very close to -1.

The R2 value as seen in the plot above is 0.9339; and this means or signifies
that the iodine value of the coconut oil can predict its corresponding
absorbance with a 93.4% accuracy and vice versa.

GROUNDNUT OIL

0.07
y = 0.0043x + 0.0286
0.06
R = 0.9638
Absorbance(233nm)

0.05

0.04

0.03

0.02

0.01

0
74 75 76 77 78 79 80 81 82
Iodine Value(mgKOH/g)

Fig 3.3 a plot of Absorbance (233nm) against Iodine value for groundnut oil

From the above Microsoft Excel plot (fig 3.3) the r-value was calculated to
give -0.8629 and this indicates a highly negative correlation coefficient. This is
because as the absorbance increased, the iodine value decreased hence an
inversely proportional relationship between both variables. Since the closer
the correlation coefficient to +1 or -1 the stronger the relationship between
the variables, with an r-value of -0.8629, it can be deduced that the

39
absorbance and the iodine value have a moderately strong linear relationship
since the r-value is quite close to -1.
The R2 value as seen in the plot above is 0.9638; and this means or signifies
that the iodine value of the groundnut oil can predict its corresponding
absorbance with a 96.4% accuracy and vice versa.

SOYABEAN OIL

0.08

0.07
y = 0.0077x + 0.0007
R = 0.9186
Absorbance(233nm)

0.06

0.05

0.04

0.03

0.02

0.01

0
124 126 128 130 132 134 136 138 140
Iodine value(mgKOH/g)

Fig 3.4 a plot of Absorbance (233nm) against Iodine value for soyabean oil

From the above plot (fig 3.4) the r-value obtained was -0.8587 and this
indicates a highly negative correlation coefficient. This is because as the
absorbance increased, the iodine value decreased hence an inversely
proportional relationship between both variables. Since the closer the
correlation coefficient to +1 or -1 the stronger the relationship between the
variables, with an r-value of -0.8587, it can be deduced that the absorbance

40
and the iodine value have a moderately strong linear relationship since the r-
value is quite close to -1.

The R2 value as seen in the plot above is 0.9186; and this means or signifies
that the iodine value of the soyabean oil can predict its corresponding
absorbance with a 91.7% accuracy and vice versa.

RELATIONSHIP BETWEEN TIME AND IODINE VALUE OF THE OIL

SAMPLES

160

140
Iodine value(mgKOH/g)

120

100
soyabean
80 groundnut oil

60 coconut oil
palm oil
40

20

0
1 3 5 7 9 11 13 15
time (days)

Fig 3.5 the plot of Iodine value (mgKOH/g) against time (days)

From the graph above, it was observed that the iodine values of the four oil
samples decreased with time.
In the period of 15 days (2 weeks), the iodine value of palm oil decreased from
52.03 to 38.77mgKOH/g i.e. a 25% decrease and this is due to its high
susceptibility to oxidation. The iodine value of groundnut oil decreased from

41
80.82 to 75.1mgKOH/g which means it experienced a 7.1% decrease. The
iodine value of soyabean oil was also observed to decrease from 138.19 to
126.29mgKOH/g hence experiencing an 8.6% decrease. Lastly, the iodine
value of coconut oil was seen to decrease from 10.63 to 8.8mgKOH/g,
experiencing a 17.22% decrease. It can be said that one of the reason for the
decrease in iodine value is due to the effect of rancidity on the oils. This
occurred due to the effect of oxidation/photo-oxidation of the oils which
caused the oils to decrease in unsaturation thereby leading to decreasing
iodine values.

RELATIONSHIP BETWEEN TIME AND ABSORBANCE OF THE OIL SAMPLES

0.8

0.7

0.6
Absorbance (233nm)

0.5
soyabean oil
0.4 groundnut oil
coconut oil
0.3
palm oil
0.2

0.1

0
1 2 3 4 5 6 7 8
time (days)

Fig 3.5 plot of Absorbance (233nm) against time (days)

From Fig 3.5 it can be seen that the absorbance of the oil samples increased
with time with palm oil showing significant increase compared to the other
samples.
42
In the duration of 15 days the absorbance of palm oil increased from 0.164 to
0.71, groundnut oil increased from 0.033 to 0.063, soyabean oil increased
from 0.016 to 0.068 and lastly coconut oil increased from 0.041 to 0.077. it
can also be observed that there was significant increase in the absorbance of
palm oil compared to the other oil samples.

43
 CONCLUSION

From the results and values obtained it was observed that the iodine values of
the oils decreased with time while their corresponding absorbance increased.

Also from the research, it was observed that the r-values of the plot of
absorbance against iodine values of all the oils used were all negative, which
means as the iodine values decreased, the absorbance of the respective oils
increased hence an inversely proportional relationship exists between both
variables. The r-values recorded for the respective absorbance vs. iodine value
plot of the different oils were; palm oil (-0.7487), coconut oil (-0.9841), g/nut
oil (-0.8629) and soyabean oil (-0.8587). These values showed a highly
negative strong linear relationship between the absorbance and the iodine
values of the oils.

The R2 values obtained from the plot of absorbance against iodine values of all
the oils were; palm oil (0.6384), coconut oil (0.9339), g/nut oil (0.9638) and
soyabean oil (0.9186). This means that the iodine value of the respective oil
can predict their corresponding absorbance with a 64%, 93.4%, 96.4%, and
92% accuracy respectively.

44
 RECOMMENDATIONS

1. Research should be conducted on the iodine values and absorbance of

other vegetable oils like linseed oil, olive oils etc.
2. The containing vessel should be opaque i.e. it should be impenetrable by
light.
3. The option of using another absorption wavelength should be explored.

45
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49
 APPENDIX

Table I Palm oil (Absorbance vs Iodine value)

Iodine Absorbance
value(mgKOH/g) (233nm)
52.03 0.164
50.41 0.209
47.2 0.21
45.75 0.238
44.03 0.245
41.69 0.259
39.34 0.381
38.77 0.71

Table II Groundnut oil (Absorbance vs Palm kernel oil)

Iodine Absorbance
value(mgKOH/g) (233nm)
80.82 0.033
78.3 0.039
76.55 0.042
76.35 0.044
76.26 0.046
75.71 0.054
75.1 0.061
75.1 0.063

50
Table III Coconut oil (Absorbance vs Palm kernel oil)

Iodine Absorbance
value(mgKOH/g) (233nm)
10.63 0.041
10.55 0.045
10.26 0.053
9.96 0.059
9.82 0.06
9.74 0.061
9.67 0.065
8.8 0.077

Table IV Soyabean oil (Absorbance vs Palm kernel oil)

Iodine Absorbance
value(mgKOH/g) (233nm)
138.19 0.016
135.43 0.018
132.76 0.019
130 0.023
128.28 0.039
127.83 0.042
126.29 0.057
126.29 0.068

51
Table V Absorbance values for the oil samples.

Day Palm Groundn Soyabean Coconut

s oil ut oil oil oil
1 0.164 0.033 0.016 0.041
3 0.209 0.039 0.018 0.045
5 0.21 0.042 0.019 0.053
7 0.238 0.044 0.023 0.059
9 0.245 0.046 0.039 0.06
11 0.259 0.054 0.042 0.061
13 0.381 0.061 0.057 0.065
15 0.71 0.063 0.068 0.077

Table VI Iodine values of the oil samples

Days Palm oil Groundnut Soyabean oil Coconut oil

oil
1 52.03 80.82 138.19 10.63
3 50.41 78.3 135.43 10.55
5 47.2 76.55 132.76 10.26
7 45.75 76.35 130 9.96
9 44.03 76.26 128.28 9.82
11 41.69 75.71 127.83 9.74
13 39.34 75.1 126.29 9.67
15 38.77 75.1 126.29 8.8

52