Characterization of lipids from oil with the use of grease-spot test, saponification

test, Acrolein test and unsaturation test

Abellar, Michael P.*, Alvarez, Audrey M., Aquino, Rachel P., Baybay, Sofia E.
3Psy2, Department of Psychology, College of science UST, Manila
Hiruha_eternal_08@yahoo.com
Abstract

Introduction
Lipids are a large and diverse group of naturally occurring organic compounds
that are related by their solubility in nonpolar organic solvents and general insolubility in
water. They also perform important functions in & both plants and animals. When
metabolized, lipids are oxidized to release large amounts of energy that are useful to
living organisms.
Fatty acids rarely occur as free molecules in nature but are usually found as
components of many complex lipid molecules such as fats (energy-storage compounds)
and phospholipids (the primary lipid components of cellular membranes. There are two
types of fatty acid, saturated and unsaturated. Saturated fatty acid is the simplest fatty
acids, they are unbranched, linear chains of CH 2 groups linked by carbon-carbon single
bonds with one terminal carboxylic acid group. Saturated means that the maximum
possible number of hydrogen atoms that are bonded to each carbon in the
molecule. The next type of fatty acid is unsaturated, it have one or more carbon-carbon
double bonds. Indicates that fewer than the maximum possible number of hydrogen
atoms that are bonded to each carbon in the molecule, it is called unsaturated.
What is saponifiable and non-saponifiable lipids?
In the experiment, the saponifiable lipids are characterized by using the
grease'spot test, saponification test and unsaturation test.

Where it is usually found, and use?

Composition of lipids.. types of lipids and define
Methodology
In characterizing lipids, the students used 4 methods which are grease-spot test,
saponification test, acrolein test and unsaturation test. The groups were asked to bring
oil for the experiment. Argan oil is the oil used by the researcher.
A. Grease-spot test
The first part of the experiment was grease-spot test, where the student where
asked to get a filter paper and divide it into 4 parts. Each part of the filter paper are
labeled with water (

H2O

), Lecithin, Dichloromethane (DCM) and the sample oil

(Argan oil). After doing the first part, the experimenter recorded the observation after
putting 2 drops of each of the solution to their respected area. The students were asked
to heat the filter paper either on a hot plate which is adjusted to its lowest setting or
place it on a watch glass over a boiling water bath, and record the appearance after
heating the filter paper.
B. Saponification Test
The group were asked to label 3 large-sized test tube with oil, fat, and

H2O

After putting labels to each of the test tube, 8 drops of oil, melted fat and water was put
to each of the test tubes. 10 mL of 3M NaOH solution was added to each of the solution
and to be heated in a boiling water bath for 15-20 minutes. After heating the test tubes,
they are to be placed in a test tube rack and let it cool to a room temperature. As the
test tube cooled, 5 mL of water was added to the solution and placed a stopper to the
test tube. The test tubes were mixed vigorously by shaking it. After mixing the solutions,
the appearance was recorded. A solution

H 2 SO 4

to acidify the solutions, it is checked

by using a blue litmus paper. The solution was mixed by a stirring rod and the material

that formed on the top part of the solution was used to check the pH and observation
were recorded.
C. Unsaturation Test
The group was asked to label 3 large-sized test tube with oil, fat, and
Glycerol. To each of the test tube, 3 mL DCM were placed to the 3 large-sized test
tubes. After putting a 3 mL DCM, the group added a 10 mL of oil, melted fat, and
glycerol. The solution was thoroughly mixed. After preparing the test tubes, the
group used a funnel to transfer the 5% bromine-dichloromethane to the 50-mL buret,
the initial volume was recorded. To each of the test tube, put 5% Brominedichloromethane dropwise while stirring the solution.until it reaches a reddish-brown
color. After the procedure, the group recorded the final volume of the solution in the
buret, and calculated the volume of the solution that is required for the reaction.
Results and Discussion
A.
B.
C.
D.

Grease-spot test
Saponification test
Unsaturated test
Acrolein test

Acrolein test was not performed during the experiment but can also be utilized to

characterize saponifiable lipids. It is performed by heating the lipid with

KHSO 4

for a

few minutes and allowing it to cool while taking note of the odor produced. When a lipid
is hydrolyzed, one of the products is glycerol. When heated with a dehydrating agent

such as potassium hydrogen sulphate,

KHSO 4

, glycerol yields acrolein, a compound

with the peculiar, acrid odor of burnt bacon was the dehydrating agent in this
dehydration reaction to produce acrolein.

Conclusion
the saponifiable lipids were characterized using the grease-spot test resulting to a
translucent spot for both lecithin and plant oils which is because of their high boiling
point, the saponification test where soaps formed and liberated fatty acids on top, and
the unsaturation test where the bromine-dichloromethane decolorized upon addition to
the lipids for both the palm oil and melted fat which qualitatively tested for the presence
of unsaturated fatty acids and relative unsaturation.

Acknowledgement
The students are thankful to their professors for guidance and for helping finish
the experiment. The groups also want to thank their classmates for providing help and
data gatherings. Lastly, the technicians for providing the materials needed for the
experiment.
References
http://www.britannica.com/science/lipid
https://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/lipids.htm