A coupled NAD+-peroxidase spectrophotometric assay for cholesterol.
We describe a modified enzymic reagent for determination of cholesterol and cholesterol ester in serum and in high-density lipoprotein. In this new procedure, the hydrogen peroxide produced by the action of cholesterol oxidase is used as a substrate for NAD+ peroxidase. Spectrophotometric determination of the NADH consumed in this coupled reaction provides a direct and absolute measure of the cholesterol originally present in free form and that liberated by the action of cholesterol ester hydrolase.
D002784:Cholesterol / Q000097:blood*; D002788:Cholesterol Esters / Q000097:blood; D002789:Cholesterol Oxidase / Q000378:metabolism; D008076:Cholesterol, HDL; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008075:Lipoproteins, HDL / Q000097:blood; D009243:NAD / Q000032:analysis / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism*; D013053:Spectrophotometry / Q000379:methods
D002788:Cholesterol Esters; D008076:Cholesterol, HDL; D008075:Lipoproteins, HDL; D009243:NAD; D002784:Cholesterol; D006861:Hydrogen Peroxide; D002789:Cholesterol Oxidase; D010544:Peroxidases; C021845:NAD+ peroxidase