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Clinical Chemistry 22: 2042-2045, 1976;
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Clinical Chemistry, Vol 22, 2042-2045, Copyright © 1976 by the American Association for Clinical Chemistry

Enzymatic Rate Method for Measuring Cholesterol in Serum

Michael A. Pesce 1 and Selma H. Bodourian 1

1 Microchemistry Laboratory, Babies Hospital, The Children's Medical and Surgical Center of New York at Columbia-Presbyterian Medical Center, 3959 Broadway, Box 54, New York, N. Y. 10032

An enzymatic rate assay is described for measuring cholesterol in serum. Cholesterol is anzymed by mixing 5 µl of sample with a reagent consistaing of cholesterol esterase, cholesterol oxidase, catalase, acetylacetone, methanol. and hydroxypolyethoxydodecane in a ammonium phosphate of the dihydrolutidine product is measured at 37 °C and 405nm. The change in absorbance between 4 and 10 min is used to calculated the cholesterol concentrations by using simultaneously determined free cholesterol standards. The change is linearly ralated to cholesterol concentration up to 4 g/liter. Samples containing bilirubin up to 200 mg/liter, uric acid up to 200 mg/liter, and hemoglobin up to 1 g/liter, or certain drugs (clofibrate, phenobarbital, nicotinic acid, salicylate, Ketochol, and Ovral) gave no interference. Ascorbic acid added to serum caused a positive interference. Lipemic samples gave values that were slightly lower than did the method of Abell et al., used for comparison. Our kinetic assay, compared with the method of Abell et al., the enzymatic assay used with Abbott's Bichromatic Analyzer, and the Technicon SMA 12/60 enzymatic procedure gave correlation coefficient of 0.992, 0.985, and 0.986, respectively.

Submitted on August 10, 1976
Accepted on September 23, 1976






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Copyright © 1976 by the American Association for Clinical Chemistry.