When the chloroform solution of cholesterol was mixed with acetic anhydride and sulfuric acid, a characteristic green or deep-blue green was produced. The intensity of the color is proportional to the amount of steroid present. The color is due to the hydroxyl group (-OH) of cholesterol reacting with the reagents a nd increasing the conjugation of the unsaturation in the adjacent fused ring. http://www.uobabylon.edu.iq/uobColeges/ad_downloads/5 _29708_805.pdf
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DISCUSSION The Liebermann-Burchard or acetic anhydride test is used for the determination of cholesterol, and in a clinical setting, it s used to determine the serum cholesterol.
1. In one test tube containing a small pinch of cholesterol, add chloroform drop by drop until it is dissolved.
2. Add 15 drops of acetic anhydride, mix and cool.
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A. LIEBERMANNBURCHARD TEST
LiebermannBurchard Test
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Liebermann-B Salkowski rea dehydrating, sulphonating reagents, sulf anhydride, to a structural for
The hydroxyl responsible Liebermann-B double bond
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drop until dissolved.
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cholestapolyene carbonium ion (iron - salt - acid reaction) . http://www.uobabylon.edu.iq/uobColeges/ad_downloads/5 _29708_805.pdf
2. Incline the tube and add slowly down the side of the test tube 5 drops of concentrated sulfuric acid.
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