JOPHAS

INTRODUCTION
Cocos nucifera is a large palm growing
up to 30 meters (98 ft) with pinnate
leaves 4-6 meters (13-20 ft) long, the old
leaves breaking away clearly leaving the
trunk smooth. (1). It takes 11-12 months
for the coconut to mature. Scientists had
identified over 60 species of coconut
palm. Today, the coconut is monotypic
Journal of
Pharmaceutical and
Allied Sciences



































with one specie Cocos nucifera. The
coconut palm is mostly found in tropics
and can be all three; a fruit, a nut, a seed.
The kernel of mature coconut is rich in
oil and in fact the most valuable part of

Journal of Pharmaceutical and Allied Sciences
Vol. 9 No. 3 (2012)
ISSN: 1596-8499
Website: http://ajol.info/index.php/jophas
COMPARATIVE PHYTOCHEMICAL SCREENING AND PHYSICO-
CHEMICAL CHARACTERIZATION OF COCONUT (Cocos nucifera)
OILS FROM TWO DIFFERENT LOCATIONS IN KADUNA STATE,
NIGERIA

*V.O. AINA, SAUDATU A. JERE, ADEWUMI A.A.J, IBRAHIM L. Y.,
YERUWAN P. ACHI, HAUWA UMAR, HAUWA HARUNA M.S. AND
ZAKARI AMINA

DEPT OF APPLIED SCIENCE, COLLEGE OF SCIENCE AND
TECHNOLOGY, KADUNA POLYTECHNIC,
P.M.B 2021 KADUNA NIGERIA
*Corresponding Author E-mail Address: vocwummi:2006@yahoo.com

ABSTRACT
The phytochemical constituents and characterization of coconut (Cocos nucifera) oils obtained
from coconut samples in two different locations in Kaduna State were investigated. The coconut
shells were removed and the fleshy parts chopped into tiny pieces, dried at room temperature
and milled into a paste using an electric blender. The oil was extracted using soxhlet
extractor/apparatus. The phytochemical constituents were determined using standard methods.
Physicochemical characterization was carried out to determine the saponification value,
refractive index, specific gravity, peroxide value and acid value of the oil. The results of the
phytochemical screening showed that alkaloid, carbohydrate, saponins, glycosides were present
in the sample obtained from Romi New Extension junction (sample 4), while flavonoids,
tannins, steroids, anthraquinones, resins and cardiac glycosides were absent. For the sample
obtained from Mando Market, flavonoids, alkaloids, steroids, carbohydrate, saponin, glycosides
were present while tannins, anthraquinones, resins and cardiac glycosides were absent.
Similarly, the results of the characterization revealed the following values for the two different
samples, respectively: saponification value (263+ 1.05, 260+ 0.01), refractive index (1.47+ 0.01,
1.478+0.002), acid value (1.00+, 1.30+0.5), specific gravity (0.84+ 0.04, 0.86+0.07) and
peroxide value (4.1+ 0.41, 3.7+0.57).

Keywords: Cocos nucifera, phytochemical properties, physic-chemical characterization.

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Aina et al/Journal of Pharmaceutical and Allied Sciences 9 (3) (2012) 1653 - 1660


the fruit, as it provides important
ingredients of food. Coconut oil is very
heat-stable, which makes it suited for
methods of cooking at high temperature
like frying. Because of its ability, it is
slow to oxidize and thus resistant to
rancidity, lasting up to two years due to
high saturated fat content (2).

Coconut palm found across the tropics is
known for its great versatility as seen in
many domestic commercial and industrial
uses of its different parts. The clear liquid
coconut water within, is a refreshing
drink and can be processed to alcohol.
The husks and leaves can be used as
material to make a variety of products for
furnishing and decorating. It has cultural
and religious significance in many
societies (1). Coconut oil has less than
90% saturated fatty acids; hence it is less
attractive to consumers.

Compared to all other oil seed crops,
coconut has the highest productivity as
well as consistency in production and is
less susceptible to abnormal climatic
conditions (3). Coconut oil has a long
shelf life and is used in baking industries,
processed foods, pharmaceutical
industries, etc. (6). Coconut oil has been
tested for use as a feed stock for biodiesel
to be used as diesel engine fuel. In this
manner, it can be applied to power
generators and transport using diesel
engines. Since straight coconut oil has a
high setting temperature (22-25
0
C), a
high viscosity, and a minimum
combustion chamber temperature of
500
0
C (932F) (to avoid polymerization of
the fuel) coconut oil is typically trans-
esterified to make biodiesel. Coconut oil
is correctly used as fuel for transport in
Philippines. Coconut oil has also been
tested for uses as engine lubricant and
transformer oil (4). Acids derived from
coconut oil can be used as herbicides (5).
Coconut oils are used for making soaps,
skin products, perfume and other personal
care and cosmetic products.

Various fractions of coconut oil are used
as drugs. Butyric acid is used to treat
cancer while lawic acid is effective in
treating viral infections. Studies carried
out on native diets high in coconut oil
show that populations consuming coconut
are generally in good health.

The incredible medicinal properties of
coconut oil are approved not only by
traditional medical practitioners but also
by modern orthodox medicine. Once
ascribed for increasing cholesterol levels,
coconut oil is now actually being used by
doctors in the treatment of a variety of
disorders. Research conducted by various
medical institutes have shown that
coconut oil possesses antimicrobial and
antiviral properties. Lauric acid is a
medium chain fatty acid which is
abundant in coconut oil and considered
responsible for many of its health benefits
(7).
The aims of the present study are:

i. To trace plant constituents
(phytochemical screening) and
determine the active constituents
of the Cocos nucifera using
standard tests to determine the
presence or absence of Alkaloids,
Flavonoids, Tannins, Steroids,
Cyanogenic glycosides,
Anthraguinones, Saponins,
Cardiac Glycosides,
Carbohydrates, Fixed oils, Fats
and Volatile oils.

ii. To characterize the extracted oil
by subjecting it to different tests
using standard methods for Acid
value, Saponification value,
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Aina et al/Journal of Pharmaceutical and Allied Sciences 9 (3) (2012) 1653 - 1660


Iodine value, Peroxide value,
Refractive index and oil content.


MATERIALS AND METHODS
This work was carried out between the
months of November, 2011 June, 2012
in the Biochemistry Laboratories of the
Departments of Applied Science and
Food Technology, Kaduna Polytechnic,
Kaduna-Nigeria.

SAMPLE COLLECTION AND
PREPARATION
The matured coconut fruits were bought
from Romi New Extension Junction and
Mando Market, all in Kaduna. The
samples were appropriately labeled and
the shells broken. The flesh was removed
from the shell, washed and rinsed with
distilled water. The samples were cut into
smaller pieces and dried at room
temperature for 5 days after which they
were mashed into paste using an electric
blender.

EXTRACTION OF OIL
Petroleum ether (250 ml) was poured into
a round bottom flask. A 100-kg quantity
of the mashed sample was weighed using
a digital weighing balance and transferred
into an extraction paper placed in the
thimble and inserted into the centre of the
extractor. The soxhlet apparatus was
heated to 40
0
60
0
C and the solvent
boiled with the vapour rising through the
vertical tube into the condenser at the top.
This was allowed to continue for 3 hours
until the extraction was complete. The
extracted oil sample was then removed
from the tube, the resulting mixture was
then subjected to distillation in order to
separate the extracted oil from the
petroleum ether. The pure (distilled) oil
was weighed so as to obtain the
percentage of oil extracted and the oil
kept for subsequent analysis.

PHYTOCHEMICL SCREENING
Phytochemical tests were carried out on
the oil extracts for the presence of
alkaloids, tannins, saponins, flavonoids,
carbohydrates, resins, anthraquinones,
steroids and glycosides according to the
methods outlined by Trease and Evans
(8), Iwu (9) and Harborn (10).

Test for saponins
The extract (0.1 ml) was shaken with 10
ml water in a test-tube for 3 minutes. The
production of frothing which persisted on
heating indicated the presence of
saponins.

Test for tannins
A small portion of the petroleum ether
extract was dissolved in 10 ml of water
and filtered. To the filtrate, 2 drops of
freshly prepared ferric chloride was
added. A dark green colour indicated the
presence of tannins.

Test for flavonoids
The extract (0.5 ml) was dissolved in 2
ml of dilute sodium hydroxide solution.
A few drops of concentrated sulphuric
acid were then added. A yellow
coloration indicates the presence of
flavonoids.

Test for resins
To 2 mls of the extract, 10 mls of acetic
anhydride was added to dissolve by
gentle heating. After cooling, 0.5 ml of
sulphuric acid was added. A bright purple
colour indicates the presence of resins.

Test for anthraquinones
The extract (0.1 ml) was dissolved in 10
mls of benzene and filtered; 5 mls 10%
ammonia solution was added to the
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Aina et al/Journal of Pharmaceutical and Allied Sciences 9 (3) (2012) 1653 - 1660


filtrate. The presence of pinkish color in
the lower ammoniacal layer signifies the
presence of anthraquinones.

Test for steroid
The extract (0.1 ml) was dissolved in
chloroform and 1 ml of concentrated
sulphuric acid was added along the side
of the test-tube to form a lower layer. A
reddish brown color at the interface
confirms the presence of steroid.

Test for reducing sugar
The extract (0.1 ml) was dissolved in 5
mls water and filtered; the filtrate was
heated with 5 mls of Fehlings solution A
and B. Formation of brick red precipitate
indicates the presence of reducing sugar.

Test for alkaloids
Two drops of Meyers reagent were
added to 0.1 ml of the extract in a test
tube. Cream precipitate indicates the
presence of alkaloids.

Test for cardiac glycosides
The extract (0.5 ml) was dissolved in 2
mls of chloroform and a few drops of
concentrated H
2
SO
4
were added to form a
lower layer. A reddish brown colour at
the interface indicates the presence of
steroidal ring.

PHYSICOCHEMICAL
CHARACTERIZATION
Determination of saponification value
The oil sample (2.0 g) was weighed and
dissolved in 2 ml petroleum ether. The
mixture was then quantitatively
transferred to 250 ml conical flask. The
beaker was rinsed 3 times with the
solvent, 25 cm
3
of 0.1 M alcoholic KOH
added and the flask attached to a reflux
condenser. Another reflux condenser was
set up as blank without oil and both flasks
were heated on a water bath for 30
minutes to boil, removed and allowed to
cool at room temperature. The mixtures
were then titrated with 0.5 M HCl using
phenolpthalein as indicator. The
difference between the blank titer value
and the mixture titer value gives the
millimeters of 0.1 M KOH required to
saponify 1 g of the oil.

Saponification value=K(V
b
V
s
) X 100
Weight of oil
Where: V
b
=Volume of HCl used in the blank
titration
V
s
=Volume of HCl used in sample
titration
K = Standard constant value = 56.1

Determination of acid value
Diethyl ether (25 ml) and ethanol (25 ml)
were mixed and 1 ml of phenolphthalein
solution (1%) was carefully added. The
mixture was neutralized with 0.1 M
NaOH. Thereafter 5.0 g of the oil was
dissolved in the mixture and titrated with
0.1M NaOH solution with constant
shaking until a pink color that persisted
for 15 seconds was obtained.

Acid Value =Titre value x M x 100
Weight of sample used

Where M = Molarity of NaOH


Determination of peroxide value
The oil sample (0.2 g) was weighed into a
conical flask, 1.0 g potassium iodide (KI)
and 20 ml of petroleum ether (solvent)
were added and mixture transferred into a
conical flask containing 20 mls of 5% KI.
Three drops of starch solution were added
and the mixture was then titrated against
0.1 M NaS
2
O
3
until the colour changed
from blue to colorless. The procedure
was then repeated for a blank titration
(without the oil).


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Aina et al/Journal of Pharmaceutical and Allied Sciences 9 (3) (2012) 1653 - 1660


Peroxide
value =Molarity x (sample titre blank titres)
Weight of sample

= M(S B) x 100
W
Where: M = Molarity of Na
2
S
2
O
3

S = Sample titre value
B = Blank titre value
W = Weight of sample

Determination of specific gravity
The specific gravity of the sample was
determined using an electronic weighing
balance. This was done by first weighing
10 cm
3
of an empty specific gravity bottle
and then weighing the bottle after 10 cm
3

of distilled water or the oil sample had
been added.

Specific gravity = Weight of oil
Weight of water
Where:
Weight of oil = Weight of oil +
bottle weight of bottle
Weight of water= Weight of water +
bottle weight of bottle

Determination of refractive index
A refractometer was used for the
measurement of the refractive index. The
oil was heated to room temperature. The
oil was allowed to drop on the plate
surface (one-two drops) after which it
was closed and the oil refractive index
determined using the eye piece with the
angle adjusted to 180
0
using the knob.

RESULTS
The results of the phytochemical
screening are presented in Table 1, while
the results for the physicochemical
characterization are presented in Table 2.

SAPONIFICATION VALUE
The analysis showed that the
saponification value of the coconut oil
(Sample A) obtained from Romi New
Extension Junction is 263+1.05 higher
than the coconut oil (Sample B) obtained
Mando Market 260+ 0.9. Both are
however within the standard reference
value range.

REFRACTIVE INDEX
Based on the results obtained, coconut oil
(Sample B) obtained from Mando market
had higher refractive index 1.478 + 0.002
than coconut oil (Sample A) obtained
from Romi New Extension Junction
1.476+ 0.01. The higher refractive index
may indicate the presence of greater
amount of impurities, which are usually
insoluble in the oil.

ACID VALUE
The result showed that the acid value of
coconut oil (Sample B) obtained from
Mando Market was higher (1.30+ 0.5)
than that for coconut oil (Sample A)
obtained from Romi New Extension
Junction (1.00+0.01). This indicated the
presence of more free fatty acids in the
former.

SPECIFIC GRAVITY
The result indicated that coconut oil
(Sample B) obtained from Mando Market
had higher specific gravity of 0.86+0.02
than coconut oil (Sample A) obtained
from Romi New Extension Junction.

PEROXIDE VALUE
Coconut oil (Sample A) obtained from
Romi New Extension Junction had higher
peroxide value of 4.1+0.41 than Coconut
oil (Sample B) obtained from Mando
market (3.7+0.57).

DISCUSSION
The results of the phytochemical
screening of the extracts of Cocos
nucifera oil are summarized in Table 1.
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Table 1: Results of the phytochemical screening of petroleum ether extracts
of coconut oil (Cocos nucifera)


CONSTITUENTS INFERENCE
EXTRACT
A B

1 Flavonoid - -
2 Tannins - -
3 Alkaloids + +
4 Steroids - +
5 Carbohydrate + +
6 Saponin + +
7 Antroquinones - -
8 Renins - -
9 Cardiac
Glycosides
- -
10 Glycosides + +

KEYS: + = Present;



- = Absent


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Aina et al/Journal of Pharmaceutical and Allied Sciences 9 (3) (2012) 1653 - 1660



Table 2: The physicochemical characterization of the various oil extracts


CHARACTERISTICS

NAME OF SAMPLES
COCONUT OIL
SAMPLE A
COCONUT OIL
SAMPLE B
Saponification Value mg KOH/g 263 + 1.05 260 + 0.91
Refractive Index Value mg KOH/g 1.476 + 0.01 1.478 + 0.002
Acid Value mg KOH/g 1.00 + 0.01 1.30 + 0.5
Specific gravity 0.84 + 0.04 0.86 + 0.02
Peroxide value meq/kg 4.1 + 0.41 3.7 + 0.57



The sign (+) indicated the presence of the
constituents, while (-) indicated the
absence of the constituents analyzed. For
sample A, coconut obtained from Romi
New Extension Junction, there were
positive results for the following
constituents: alkaloids, carbohydrates,
saponin, glycosides and negative results
for flavonoids, tannins, steroids and
anthraquinones, resins and cardiac
glycosides. For sample B, coconut oil
obtained from Mando Market the results
for the following constituents were
positive: flavonoids, alkaloids, steroids,
carbohydrate, saponin, glycosides and
negative for tannins, anthraquinones,
resins and cardiac glycosides.

From the results shown in Table 2, the
coconut oil obtained from Romi New
Extension junction had a higher













saponification value than the sample
obtained from Mando Market. The two
values were however within the specified
ranges for vegetable oils (11).


Refractive index is a physical
characteristic of triglycerides determined
by the angle through which a beam of
light is refracted or bent when passing
through a thin film or melted fat. It is
usually used in testing the purity of the
component of mixture. From Table 2,
there was a higher value of refractive
index for the coconut oil obtained from
Mando Market than that obtained from
Romi New Extension junction. This may
be as a result of contamination during the
exposure of the oil to air or during the
process of processing the oil.











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Aina et al/Journal of Pharmaceutical and Allied Sciences 9 (3) (2012) 1653 - 1660


Some of the deteriorations that occur
during storage of either the raw material
from which the fat or oil is produced or in
the fat itself after extraction lead to
hydrolysis of triglycerides to yield free
fatty acids. Acid value is used as a
general indication of the condition and
edibility of oils and measures the amount
of free fatty acids, present in oils or fats.
Table 2 shows that coconut oil obtained
from Mando Market had higher acid
value than that obtained from Romi New
Extension junction.

The result of specific gravity obtained
showed that coconut oil obtained from
Mando Market had higher value than that
obtained from Romi New Extension
junction with values within the range of
the standard reference values. Also,
peroxide value is usually used as an
indicator of the deterioration of fats and
oils. It is the measure of oxygen on the
unsaturated bonds and consequently, the
oxidative reaction results in the formation
of volatile products, loss of taste and
odour resulting in odour free rancidity.
From Table 2, therefore the coconut oil
obtained from Romi New Extension
Junction had higher peroxide value than
that obtained from Mando Market,
showing more stability of the later.
According to Pearson (12), the accepted
standard of peroxides value is below 10
meg/kg and indicate the freshness of fats
or oil with rancidity being noticed at
peroxide values between 20-40 meg/kg.


ACKNOWLEDGEMENT
The authors wish to express their
gratitude to the Head of Department,
Applied Sciences Department, Kaduna
Polytechnic, Kaduna Nigeria for

granting access to the departmental
laboratories and reagents.

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