How Does the Liebermann-Burchard Test Determine a Cholesterol Level?

The Liebermann-Burchard test involves observing the reaction of cholesterol dissolved in chloroform with acetic anhydride and two drops of concentrated sulphuric acid. According to Wieland and Weil, the reaction results in the formation of a green-blue colored product, a sulphonic acid derivative of cholesterilene, which indicates the presence of cholesterol.

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.