Enzymatic Determination of Total Serum Cholesterol
1974/04/01 by Charles C Allain, Charles C. Allain, Lucy S Poon +5 · 33 citations
Biochemistry, Genetics and Molecular Biology · Chemistry · #Steroid Chemistry and Biochemistry #Receptor Mechanisms and Signaling #Mass Spectrometry Techniques and Applications
paper · pdf · doi:10.1093/clinchem/20.4.470
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.
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