Soran University – Faculty of Science

Chemistry department
Biochemistry
3rd stage – Group (A)

Experiment No. (5):

Qualitative analysis of Lipids part I

Prepared by:

Zheno Ali
Azhy Azad
Yusra Rzgar
Rawen
Hassan Sima
Sherzad
Belal Muhammad
Introduction

Solubility test

The ability of a compound to dissolve in a liquid, which works as a solvent helps in determining
the solubility test. The tests are so essential because they identify the polarity of the unknown
compounds. It also helps in determining the presence of acidic and basic functional groups. The
highest concentration of solute can be measured through this test. In this experiment to detect
the presence of lipid with Chloroform, Ethanol, Water.

Translucent Spot Test

In this, test the given sample to be tested is rubbed between the folds of filter paper. The
appearance of translucent spot confirms the presence of fats in the given sample. If there is a
presence of translucent spot then the presence of fats is confirmed.

Acrolein test

The simplest unsaturated aldehyde is acrolein (propenal in scientific terms). It's a colorless
liquid with an acrid, piercing odor. The scent of burnt fat is caused by glycerol in the burning
fats breaking down into acrolein’s (similar to how the cooking oil is heated to its smoke point).
It's made in a factory from propylene and is mainly used as a biocide and a building block for
other chemicals.

One of the applications of Acrolein is Acrolein test is used to detect the presence of glycerol or
fat. When fat is treated strongly in the presence of a dehydrating agent like potassium bisulfate
(KHSO4), the glycerol portion of the molecule is dehydrated to form an unsaturated aldehyde,
acrolein that has a pungent irritating odor.
Procedure & Material
Material and equipment
 Water
 Hot plate
 Oil
 Test tubes
 Ethanol
 Beaker
 Acetone
 filter paper
 Chloroform
 Sodium bisulfate
 Potassium bisulfate

Procedure
1. Solubility test
I. Take the sample of lipid (1 mL) in three different test tubes by labelling it as A, B and C.
II. Then add different solvents (1 mL) like Water, Ethanol and Chloroform in the three
different test tubes.
III. 3.Shake the tubes and allow it to stand for 1minute.

2. Translucent spot test

I. Take a filter paper.
II. Add one drop of water on one end and a drop of Oil or lipid on the other end.

3. Acrolein test
I. Take 0.5 g of powdered Sodium bisulfate (NaHSO4) or Potassium bisulfate (KHSO4) in a
clean and dry test tube
II. Add 10 drops of the sample
III. Mix thoroughly and heat it using a water bath.
Discussion:
Solubility test

The purpose of this qualitative test is to know the solubility of lipids into different
solvents. In this experiment we prepared three test tube and we put oil into all of them, we
added ethanol as solvent into A test tube and water into B test tube and chloroform into C
test tube. We notice that that A test tube is partially positive, ethanol having poler and non
poler molecules, the ethyl group in ethanol can mixed with non poler molecules like lipids.
In B test tube we notice that they immiscible into each other and two layers composed,
because lipids is non poler and H2O is poler for that they cannot be miscible into each
other, the result become negative. The result of C test tube is positive because they mixed
into each other and one layer is composed. Both lipids and chloroform is non poler
because of that they can dissolved each other very well.

translucent spot Test

The aim in this preliminary test for lipid to detect the presence of lipid by appearance of
translucent spot or greasy spot .

When added the one or more drops of lipid onto the filter paper, the filter paper was not
wet and the lipid formed a spot ,the spot appeared as a translucent spot or (greasy spot )
and diffraction occurred on the lipid spot , the light photons can pass from one side to
other side of the filter paper, but when added some drops of water onto the filter paper,the
filter paper will wet, and the water didn’t form any spot on the filter paper .

In the end we observed that the appearance of translucent spot (greasy spot) on the filter
paper that mean positive result and if the translucent spot disappear on the filter paper
means negative result.
Acrolein test

The aim of this test the same with other test to detect the presence of lipid in the sample
and we obtained Acrolin or not .

In this test we heated lipid (oil or fat )because of heat the lipid hydrolysis to glycerol and
fatty acid this glycerol reacted with sodium bisulfate (NaHSO4)or potassium
bisulfate(KHSO4) that dehydrating agent to formed unsaturated aldehyde (Acrolein) .
In the laboratory we brought two unknown sample take in to test tube and add NaHSO4 or
KHSO4 and put in to the water bath heated if there is a pungent irritating odour then the
presence of lipid (fats or oil )is confirmed that mean positive result .

This test just to detect simple lipid and complex lipid not to detect derived lipid
because derived lipid not contains ester bond so that glycerol didn’t performed that’s
why didn’t obtained Acrolein mean this test to derived lipid Negative result .

In general if the lipid we used was hydrolysis lipid » positive test

None hydrolysis lipid »Negative test .

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