HPLC analysis of desmosterol, 7-dehydrocholesterol, and cholesterol.
A simple and sensitive method to analyze mixtures of desmosterol, 7-dehydrocholesterol and cholesterol is described. The method involves the oxidative conversion of the sterols with cholesterol oxidase, followed by high performance liquid chromatographic (HPLC) analysis. A C18 reversed phase column (3 microns, 75 X 4.6 mm) and a mixture of methanol and acetonitrile (1:1, v/v) at a rate of 1 ml/min are used to separate the sterols. The eluted sterols are quantified by measuring UV absorption at 240 nm. As little as 10 pmoles of sterol can be quantified under these conditions.
Medical Sciences Program, Indiana University School of Medicine, Bloomington 47405.
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D002775:Cholestadienols / Q000032:analysis*; D002784:Cholesterol / Q000032:analysis* / Q000097:blood; D002789:Cholesterol Oxidase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003684:Dehydrocholesterols / Q000032:analysis*; D003897:Desmosterol / Q000032:analysis*; D006801:Humans
D002775:Cholestadienols; D003684:Dehydrocholesterols; D003897:Desmosterol; D002784:Cholesterol; C016705:7-dehydrocholesterol; D002789:Cholesterol Oxidase