The reaction Liebermann-Burchard is a chemical reaction mainly used as a litmus test

of cholesterol and more generally phenols .

It was first described by Carl Liebermann 1 then applied to the revelation of Burchard
cholesterol by 2 , 3 .
Reagents: solution of the substance to be identified in the chloroform + sulfuric
acid concentrate + acetic anhydride

Liebermann-Burchard reagent, as prepared by Liebermann and Burchard, is

used in a chemistry laboratory to detect cholesterol. The green-blue color of the
product is due to the interaction of the hydroxyl group of cholesterol with the
reagent. This test is used for detecting unsaturated steroids and triterpenes, and
observing the reaction allows researchers to make qualitative and quantitative
estimations of cholesterol levels, according to Academic's medical dictionary.

a few drops of acetic anhydride were added to the then introduced drop a solution of
sulfuric acid.

The solution turns deep red, then purple, blue and finally takes a dark green color.

This color changing rapidly with time making it even more difficult the specific
highlight definition.

In the observation of visible color reaction of the lipids for the

Liebermann-Burchardtest both the standard lipid cholesterol and the
cholesterol extracted from the fish brain were observed to have given the
positive reaction for such test.
The Lieberman-Burchard oracetic anhydride test is performed for the
detection of the presence of sterols
This statement was taken from Lehninger verbatim
For the positive visible result, the formation of a green or blue green color
as the solution is subjected to the reagents is considered. Included reagents in
the Lieberman-Burchard test are acetic anhydride and conc. H2SO4.
The test is used in a colorimetric test to detect cholesterol, which gives a deep
green color. Thiscolor begins as a purplish, pink color and progresses through to
a light green then very darkgreen 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. Furthermore,
the principle for such reactions involves the esterification of OH at carbon 3
with acetic anhydride and also the epimerization of double bond at carbon 5.
This statement was taken from Boyer verbatim.

Cholesterol is by far the most common member of a group of steroids in

animal tissues;it has a tetracyclic ring system with a double bond in one of
the rings and one free hydroxylgroup. It is found both in the free state,
where it has an essential role in maintaining membranefluidity, and in
esterified form, i.e. as cholesterol esters