1. Inventi Rapid: Pharm Analysis & Quality Assurance Vol. 2015, Issue 4
[ISSN 0976-3813]
2015 ppaqa 17091 © Inventi Journals (P) Ltd
Published on Web 24/08/2015, www.inventi.in
RESEARCH ARTICLE
INTRODUCTION
Sterols have been coined from the words steroidal alcohols.
They have been found to occur both in animal and plant oil
and fats. [1] These are usually crystalline and mostly bear an
alcoholic group. They may occur either as free or as esters
of the higher fatty acids and are isolated from the
unsaponifiable fraction of oils and fats. Sterols may be
further subdivided into the following three categories:
Zoosterol (sterols obtained from animal kingdom only eg:
cholesterol), Phytosterol (sterols are derived exclusively
from the plant source eg: ergosterol, stigmasterol),
Mycosterols (sterols obtained from either yeast or fungi).
Sterols are the derived lipids which are inorganic
substances insoluble in water but soluble in organic
solvents like chloroform, ether benzene.
Cholesterol was first isolated from gall stones it was
called cholesterol which means solid alcohol from bile it is
a white crystalline substance showing the usual solubility
properties of lipids it has the melting point of 149°C. [2]
Human body has two sources of cholesterol one from diet
and other by synthesis from acetate. [3] The greater the
dietary intake of cholesterol lowers the rate of cholesterol
bio synthesis in the liver and adrenal cortex. Tissues
involved in the biosynthesis of cholesterol are liver,
adrenalcortex, skin, intestine, testis and aorta. The various
diseases associated with higher intake of sterols was
atherosclerosis, coronary heart disease,
hypercholesterlemia. [4] Atherosclerosis may be defined as
degenerative changes in the intima of medium and large
arteries. [5] Degeneration includes the accumulation of
lipids, complex carbohydrates blood and blood products
and is accompanied by the formation of fibrous tissue and
calcium decomposition on the intima of the blood values.
As the total cholesterol level increases the risk of coronary
artery disease begin to rise, when total cholesterol is above
200 mg/dl the risk of a heart attack doubles with every 50
mg/dl increases in total cholesterol. [6] Natural dietary
intake varies from about 167-437 mg/day.
Consuming of saturated fats such as palm oil,
hydrogenated oil, coconut oil raises the cholesterol level
1Creative Educational Society's College of Pharmacy, Chinnatekur, Kurnool
- 518218, Andhra Pradesh, India.
E-mail: raji12pharm@gmail.com
*Corresponding author
where as the vegetable oils (poly unsaturated fats) like
sunflower oil, mustard oil etc. it is also done by taking
monosaturated fats like olive oil. The most widely used
cholesterol lowering agents in cardio vascular medicine
today are statins (Lovastatin), colestipol, nicotinic acid and
cholestyramine.[7]
For the estimation of sterols in edible oil and ghee
samples using UV– spectrophotometer by Liberman –
Burchard method was carried out. In this reaction the
sterol in the sample reacts with Acetic anhydride in the
Liberman –Burchard regent which gives a Green colour,
whose absorbance can be determined by UV-visible
spectrophotometer. [8]
MATERIALS AND METHODS
All the chemicals used in the research is manufactured by
S.D. FINE Chemicals limited, Mumbai procured from Prince
Scientific and Surgicals, 4-1-10/1, Tilak Road, Opp Church
Building, Hyderabad, A. P. India and brand and matrix
scientific.
Instrument
UV – Visible Spectrophotometer 3000+ (LAB INDIA),
Working mode: Photometry at λmax
640 nm.
Standard Cholesterol Solution
Standard cholesterol of 10 mg dissolved in 10 ml
chloroform, shake well the solution.
Liberman - Burchard Reagent
Sulphuric acid of 0.5 ml dissolved in 10 ml of acetic
anhydride in a beaker and cover the beaker with black
carbon paper and kept in ice bucket.
Sample Collection and Preparation
Ghee samples and edible oil samples were purchased from
local market at Kurnool. The different edible oil and ghee
samples include Ragava Palm Swathi (refined palm oil),
Kachi Ghani (Mustard oil), Naturrale (sunflower oil), K.S
Brand (Gingelly oil), Parachute (coconut oil), Durga (pure
ghee),Vandana (pure ghee) and Himaneesdevee (ghee)
purchased from local market Kurnool, Andhrapradesh,
India &labelled as manufacturer A, B, C, D, E, F, G and H
respectively. Weigh 1 gm of sample which is dissolved in
Estimation of Sterols Content in Edible Oil and Ghee Samples
T Raj Kumar1, V Rajeswari1*, R Meenakshi1, L Siva Sankar Reddy1, Y Siva Rami Reddy1, E
Pushpalatha1
Abstract: The present investigation was carried out for the estimation of sterol content in marketed edible oil and ghee samples.
Eight samples of edible oils and ghee samples were subjected for sterol estimation. The samples include Ragava Palm Swathi
(refined palm oil), Kachi Ghani (Mustard oil), Naturrale (sunflower oil), K.S Brand (Gingelly oil), Parachute (coconut oil), Durga
(pure ghee), Vandana (pure ghee) and Himaneesdevee (ghee) purchased from local market Kurnool, Andhrapradesh, India. The
determination was done by measuring the absorbance on UV spectrophotometer using Liberman – Burchard reaction. The
sterol contents of include Ragava Palm Swathi, Kachi Ghani, Naturrale, K.S Brand, Parachute, determined as 18 mg/gm, 48 mg/g,
56 mg/gm, 84 mg/gm and 13 mg/gm respectively, while for Durga, Vandana and Himaneesdevee found as 30 mg/gm, 29
mg/gm, 23 mg/gm. The more amounts of sterols was found to be in gingelly oil and coconut oil have less in comparison.
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2. Inventi Rapid: Pharm Analysis & Quality Assurance Vol. 2015, Issue 4
[ISSN 0976-3813]
2015 ppaqa 17091 © Inventi Journals (P) Ltd
Published on Web 24/08/2015, www.inventi.in
RESEARCH ARTICLE
chloroform to 10 ml of chloroform and this solution is
further diluted to 10 times. From this solution pipette out
3ml of the solution to this add 2 ml of Liberman-Burchard
reagent and add 2 ml of chloroform. Then the tubes of the
solution covered with the black carbon paper and place in
an ice bath for 15 min for 15 min in a dark place their
absorbance were determined on spectrophotometer at 640
nm.
Standard Cholesterol Procedure
0.5, 1.0, 1.5, 2.0, 2.5 ml of standard cholesterol taken in five
test tubes. Taken another test tube which is as blank test
tube and the test tubes were marked as S1, S2, S3, S4, S5
and S6. Then to all the test tubes 2 ml of the Liberman-
Burchard regent was added and made up the final volume
and was made equal in each test tube by adding
chloroform as shown in Table 1. Then all test tubes were
covered with carbon black paper and kept in dark place
for 15 minutes in an ice bucket. Then the base line was
taken by using blank solution, then the absorbances of all
standard solutions were determined and the graph was
plotted.
RESULTS AND DISCUSSION
The Sterol Content of edible oils of different brands like
Ragava Palm Swathi (refined palm oil), Kachi Ghani
(Mustard oil), Naturrale (sunflower oil), K.S Brand
(Gingelly oil), Parachute (coconut oil), Durga (pure ghee),
Vandana (pure ghee) and Himaneesdevee (ghee) was
determined. These determination was done by measuring
the absorbance on UV spectrophotometer at 640 nm using
Liberman – Burchard reaction. The sterol content of Ragava
Palm Swathi, Kachi Ghani, Naturrale, K.S Brand , Parachute,
determined as to be 18 mg/gm, 48 mg/gm, 56 mg/gm 84
mg/gm and 13 mg/gm respectively, where as sterol
content of ghee samples includes Durga, Vandana and
Himaneesdevee found as 30 mg/gm, 29 mg/gm, 23 mg/gm.
The more amounts of sterols found in gingelly oil and
coconut oil have less in comparisons.
CONCLUSION
The present investigation is an overview of how food
choices can affect one’s cholesterol level. Cholesterol levels
above 150 mg, the risk of heart disease increases [9] such as
coronary artery disease, atherosclerosis
hypercholestrolemia. Cholesterol in food increases the level
of LDLS.
[10] One study shows that people who adopt a
vegetarian diet reduce their fat intake by 26% and achieve
a significant drop in cholesterol levels in just six weeks. [11-
13] Beside the very low level of fat eaten in a typical
vegetarian diet, vegetables proteins also helps decrease
risk of heart attack.
Table 1: Estimation of Sterols in Edible Oils and Ghee Samplesstample1
Standard Sample
I II III Avg±S.D. (n=3) I II III
Avg±S.D.
(n=3)
% Content
of Sterol
S1 0.043 0.043 0.042 0.042± 0.0005 MfgA 0.2 0.17 0.19 0.18±0.152 18 mg/gm
S2 0.123 0.123 0.122 0.122± 0.0005 MfgB 0.5 0.49 0.46 0.48 ±0.020 48 mg/gm
S3 0.251 0.251 0.250 0.250± 0.0005 MfgC 0.7 0.4 0.6 0.56± 0.15 56mg/gm
S4 0.335 0.334 0.335 0.334 ±0.0005 MfgD 0.8 0.89 0.85 0.84 ±0.04 84mg/gm
S5 0.496 0.494 0.494 0.494± 0.0011 MfgE 0.14 0.14 0.13 0.13 ±0.005 13 mg/gm
- - - - - MfgF 0.31 0.30 0.30 0.30 ±0.008 30 mg/gm
- - - - - MfgG 0.30 0.29 0.28 0.29 ±0.013 29 mg/gm
- - - - - MfgH 0.26 0.24 0.20 0.23 ±0.03 23 mg/gm
S.D. = standard deviation, MfgA =Ragava Palm Swathi (refined palm oil), MfgB =Kachi Ghani(Mustard oil), Mfg c = Naturrale (sunflower oil), Mfg D = K.S
Brand (Gingelly oil), Mfg E= Parachute (coconut oil), Mfg F= Durga(pure ghee), Mfg G= Vandana(pure ghee) Mfg H= Himaneesdevee (ghee)
Figure 1: Estimation of sterols in edible oils and ghee samples
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3. Inventi Rapid: Pharm Analysis & Quality Assurance Vol. 2015, Issue 4
[ISSN 0976-3813]
2015 ppaqa 17091 © Inventi Journals (P) Ltd
Published on Web 24/08/2015, www.inventi.in
RESEARCH ARTICLE
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Cite this article as: T Raj Kumar, V Rajeswari, R
Meenakshi et al. Estimation of Sterols Content in Edible
Oil and Ghee Samples. Inventi Rapid: Pharm Analysis &
Quality Assurance, 2015(4):1-3, 2015.
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