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Colorimetric microdetermination of blood cholesterol

, : Colorimetric microdetermination of blood cholesterol. An Farm Y Bioquim [Buenos Aires] 11(1): 8-16

Cholesterol in 2 ml. of blood is extracted with 20 ml. of acetone at 70-75[degree] C, and an aliquot equivalent to 1 ml. of blood is filtered and dried. The residue, dissolved and made up to 5 ml. with chloroform is treated with 2 ml. acetic anhydride and then with 2 ml. H2SO4. Maximum green color of the Liebermann-Burchard reaction develops in 5-8 mins. in darkness. Standard soln. for colorimetric comparison contains 1 mg. cholesterol per ml., dissolved in chloroform and similarly treated with acetic anhydride and H2SO4. A similar micro-method using 0.2 ml. of blood is also described. Acetone extraction is done in conical centrifuge tubes. After centri-fuging the decanted soln. is treated as in the macro-method, but with correspondingly smaller amts. of reagents, and compared with more dilute standard solns.


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