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Clinical Chemistry 20: 470-475, 1974;
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Clinical Chemistry, Vol 20, 470-475, Copyright © 1974 by the American Association for Clinical Chemistry

Enzymatic Determination of Total Serum Cholesterol

Charles C. Allain 1, Lucy S. Poon 1, Cicely S. G. Chan 1, W. Richmond 2, and Paul C. Fu 3

1 Instrument/Reagents Group, Abbott Diagnostics Division, 820 Mission St., South Pasadena, Calif. 91030.
2 Clinical Research Centre, Harrow, Middlesex, England.
3 Department of Pathology, Harbor General Hospital, Torrance, Calif. 90509.

An enzymatic method is described for determination of total serum cholesterol by use of a single aqueous reagent. The method requires no prior treatment of sample and the calibration curve is linear to 600 mg/dl. Cholesterol esters are hydrolyzed to free cholesterol by cholesterol ester hydrolase (EC 3.1.1.13). The free cholesterol produced is oxidized by cholesterol oxidase to cholest-4-en-3-one with the simultaneous production of hydrogen peroxide, which oxidatively couples with 4-aminoantipyrine and phenol in the presence of peroxidase to yield a chromogen with maximum absorption at 500 nm. The method is reproducible, and the results correlate well with those obtained by automated Liebermann—Burchard procedures (AA-2 and SMA 12/60) and the method of Abell et al. The present method affords better specificity than those previously reported and has excellent precision.

Submitted on January 8, 1974
Accepted on January 31, 1974




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