Libermaann-Burchard Test  Or Acetic Anhydride Test

Libermaann-Burchard Test  Or Acetic Anhydride Test

(For the qualitative detection of cholesterol)

Principle:

The Liebermann–Burchard  test is used for the detection of cholesterol. It is also known as acetic anhydride test. To a solution of cholesterol in chloroform, when a few drops of acetic anhydride is added, mixed well, and a few drops of concentrated sulphuric acid added from side of the tube, kept in dark. A red rose colour develops which changes rapidly to deep green. One molecule of water is removed from cholesterol, and it is oxidized giving rise to green fluorescent colour.

Reagent:

  • Cholesterol solution
  • Chloroform
  • Acetic anhydride
  • Concentrated sulphuric acid

Sample preparation:

  • Separate the egg yolk (yellow part) of a boiled egg and grind it with about 50 ml of chloroform in a mortar.
  • Filter the suspension through a filter paper.
  • The filtrate is the cholesterol solution. The egg yolk is the rich source of cholesterol.

Procedure:

  • Dispense about 2 ml of cholesterol solution in dry test tube.
  • Add 10 drops of acetic anhydride and mix well.
  • Then, add 2 drops of conc. sulphuric acid from side of tube wall.
  • Keep it in dark for about 5 mins, and observe the appearance of green colour.

Source: https://www.researchgate.net/profile/Dr_Alok_Nahata

Interpretation:

Appearance of green colour indicates presence of cholesterol in solution.

Note:

Sulphuric acid and acetic anhydride are corrosive and hazardous

 

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