Enzymatic Determination of Total Serum Cholesterol

Clinical Chemistry - Tập 20 Số 4 - Trang 470-475 - 1974
Charles C. Allain1, Lucy S Poon1, Cicely S G Chan1, W. Richmond2, Paul Fu3
1Instrument/Reagents Group, Abbott Diagnostics Division, 820 Mission St., South Pasadena, Calif. 91030
2Clinical Research Centre, Harrow, Middlesex, England
3Department of Pathology, Harbor General Hospital, Torrance, Calif. 90509

Tóm tắt

Abstract 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.

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