For the quantitative determination of Total Cholesterol in serum on Hitachi analyzers.
Cholesterol esters in the sample are hydrolyzed by cholesterol esterase to release free cholesterol. The free cholesterol is oxidized by cholesterol oxidase to release hydrogen peroxide. Phenol + 4-AAP + hydrogen peroxide, in the presence of peroxidase produces a quinoneimine dye that is measured at 500nm. The absorbance at 500nm is proportional to the concentration of cholesterol in the sample.