Lipids Determination

A large number of heterogenous compounds are referred to as lipids

including fats, steroids, waxes, and related compounds, which are
related more by their physical than their chemical properties.

Qualitative Tests:
I. Physical Test:
Grease spot test
Take a small amount of oil on a piece of paper, a greasy spot
penetrating the paper will be formed. This happens because lipid does
not wet paper unlike water.

Emulsification:
Oil or liquid fat becomes finely divided and is dispersed in water when
shaken with water to form emulsification. Emulsification is permanent
and complete in the presence of emulsifying agent. The important
emulsifying agents are bile salts, proteins, soaps, mono- and
diglycerides. Emulsification is important in the processes of fat
digestion in the intestine. Emulsifying agents lower surface tension of
the liquid.

Take 2 clean and dry test tubes, in one test tube added 2 ml water and
in other 2ml dilute bile salt solution. Now to each tube added 2 drops
of mustard oil and shaken vigorously for about one minute. Allow the
tubes to stands for two minutes and note that the water, oil is broken
in small pieces and floats on the surface; where as in the bile salt
solution, the oil can be seen in minute droplets suspended in the liquid
(permanent emulsification).

The emulsion test is a method to determine the presence of lipids using wet chemistry. The

procedure is for the sample to be suspended in ethanol, allowing lipids present to dissolve (lipids
are soluble in alcohols). The liquid (alcohol with dissolved fat) is then decanted into water. Since
lipids do not dissolve in water while ethanol does, when the ethanol is diluted, it falls out of the
solution to give a cloudy white emulsion
Acrolein test
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 bisulphate (KHSO4),
the glycerol portion of the molecule is dehydrated to form an unsaturated
aldehyde, acrolein that has a pungent irritating odour