Clinical Chemistry
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Clinical Chemistry 19: 1128-1134, 1973;
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Clinical Chemistry, Vol 19, 1128-1134, Copyright © 1973 by the American Association for Clinical Chemistry

Kinetics of the Cholesterol-Sulfuric Acid Reaction: A Fast Kinetic Method for Serum Cholesterol

Thomas E. Hewitt 1 and Harry L. Pardue 1

1 Department of Chemistry, Purdue University, Lafayette, Ind. 47907.

We studied the kinetics of the reaction between cholesterol and sulfuric acid in acetic acid-acetic anhydride medium. Results have been used to establish near-optimal conditions for the fast kinetic determination of cholesterol in serum. The reaction rate measured during the first 20 s of the reaction is proportional to cholesterol concentration. There is good agreement (<2% deviation) between pseudo-first-order rate constants for cholesterol standards and sera. Recoveries of standard cholesterol added to sera range from 99% to 104% (average, 100.7%). Values for serum cholesterol by this kinetic determination tend to be somewhat lower than equilibrium values reported by local hospital laboratories. Whether bilirubin interferes depends on reaction conditions; under optimal conditions, each milligram of bilirubin is kinetically equivalent to about 1 mg of apparent cholesterol. Kinetic data are included to show how results are degraded by operating under nonoptimal conditions, and considerations involved in changing experimental conditions are discussed.


Key Words: optimal analytical conditions • bilirubin interference • analytical implications of kinetic data • Liebermann-Burchard reaction • molar absorptivity • equilibrium and kinetic values compared

Submitted on June 6, 1973
Accepted on July 10, 1973






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Copyright © 1973 by the American Association for Clinical Chemistry.