QUALITATIVE AND

QUANTITATIVE TESTS FOR

LIPIDS.
Lipids
 A large number of heterogeneous compounds
are referred to as lipids including fats,
steroids, waxes, and related compounds,
which are related more by their physical than
their chemical properties.
 They have the common property of being:

1. relatively insoluble in water

2.soluble in polar solvents such as ether
and chloroform.
Qualitative and Quantitative Tests
for Lipids
Solubility Test
Spotting Test
Acrolein Test
Saponification Test
Liebermann-Burchard Reaction for
Steroids
Salkowski’s Test (H2SO4 Test)
Overview
Lipids are water-insoluble organic compounds that can be
extracted from cells and tissues by non-polar solvents like ether,
chloroform, benzene, etc. Lipids that contain fatty acids are
saponifiable, while those without fatty acids like steroids are non-
saponifiable which do not produce fatty acid nor alcohol.
Some
common lipids are the oil, fats, wax, steroid, hormones
and
vitamins like A, D, E and K. In the body, lipids are converted to
fatty acid and glycerol by bile then by steapsin, or lipase.

In the laboratory, fats and oil may be tested qualitative

to
determine their characteristics structure like acrolein, an
unsaturated aldehyde, cholesterol, a sterol and the polar and on
polar group of saponified fatty acid.
Solubility Test
 The test is based on the
property of solubility of
lipids in organic solvents
and insolubility in water.

 PRINCIPLE: The oil will float

on water because of lesser
specific gravity.

 TEST: Take 3ml of solvents in

each test tube and add 5
drops of sample. For water
and ethanol, it is insoluble
and for chloroform and ether,
it is soluble and hence the
given sample is lipid.
Spotting
Effect
All the lipids are greasy in
nature. Therefore the test
may be taken as group
test for lipids.

 PRINCIPLE: The oil does not

wet the paper.

 TEST: Take 3ml of ether in a

test tube and dissolve 5
drops of oil in tit. Put a drop
of the solution on the filter
paper and let it dry. A
translucent spot on the filter
paper was observed and this
indicates the greasy
character of the lipid.
Acrolein Test
 used to detect the
presence of glycerol
or fat. When fat is
treated strongly in the
presence of a
dehydrating agent like
potassium bisulphate
(KHSO4), the
glycerol portion of
the molecule is
dehydrated to form an
unsaturated
aldehyde, acrolein
that has a pungent
irritating odor.
Saponification-The process of Making
Soap
 The Theory
Soaps are made from fats
and oils or their fatty
acids. The saponification
reaction is exothermic in
nature, because heat is
liberated during the
process. The soap formed
remains in suspension
form in the mixture. Soap
is precipitated as a solid
from the suspension by
adding common salt to
the suspension. This
process is called Salting
out of Soap.
Liebermann-Burchard Test (Test for Sterol)

 This test is used for the detection

of cholesterol. The formation of a
green or green-blue color after a
few minutes is positive.

 Lieberman–Burchard is a reagent
used in a colorimetric test to detect
cholesterol, which gives a deep
green color. This color begins as a
purplish, pink color and progresses
through to a light green then very
dark green color. The color is due to
the hydroxyl group (-OH) of
cholesterol reacting with the
reagents and increasing the
conjugation of the un-saturation in
the adjacent fused ring. Since this
test uses acetic anhydride and
sulfuric acid as reagents, caution
must be exercised so as not to
Salkowski Test: Test for
Cholesterol
 Dissolve cholesterol
in 2 ml of
chloroform in dry
test tube. Add equal
amount of con.
H2SO4. Shake
gently. The upper
layer turns red and
the sulphuric acid
layer shows a
yellow colour with a
References:
 https://www.biochemden.com/color-reactions
- of-lipids/
 http://www.biologydiscussion.com/lipids/test
s/q ualitative-and-quantitative-tests-for-
lipids/13050
 https://steemit.com/steemiteducation/@chamu
diliyanage/chemical-basis-of-life-part-2-let-s-

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