Abstract

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.

Keywords

Cholesterol oxidaseChemistryCholesterolHydrogen peroxideChromatographyReagentHydrolysisCalibration curvePhenolEnzymePeroxidaseBiochemistryDetection limitOrganic chemistry

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Publication Info

Year
1974
Type
article
Volume
20
Issue
4
Pages
470-475
Citations
9058
Access
Closed

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Charles C. Allain, Lucy S Poon, Cicely S G Chan et al. (1974). Enzymatic Determination of Total Serum Cholesterol. Clinical Chemistry , 20 (4) , 470-475. https://doi.org/10.1093/clinchem/20.4.470

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DOI
10.1093/clinchem/20.4.470