Abstract
A fluorometric method for the enzymatic determination of cholesterol in serum was developed. It involves the use of resorufin and peroxidase for the measurement of hydrogen peroxide generated when cholesterol oxidase acts upon cholesterol. The ratio of serum to reagent was 1:1999 which permitted the determination of cholesterol concentrations of between 0.8 and 10 μmol/liter in the final solution. The within-run and between-run CVs for two control sera containing 3.19 and 5.66% mmol/liter cholesterol were 1.9 and 2.6%, and 0.88 and 1.22%, respectively. Interferences from triglycerides up to 10.25 mmol/liter and hemoglobin up to 5.6 g/liter were insignificant, whereas the bilirubin interference is significant starting from 115 μmol/liter. Recoveries for between 2 and 6 mmol/liter standard additions to a pooled serum containing 5.87 mmol/liter were between 94.5 and 104.3%. Serum samples were analyzed using a modified Trinder method and the proposed method. The regression equation was Y = 1.02 X − 0.13 ( r = 0.98, N = 56). The proposed method for the determination of cholesterol is simple, reliable, and can be easily automated.
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