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HPLC analysis of desmosterol, 7‐dehydrocholesterol, and cholesterol
Author(s) -
Goh Edward H.,
Colles Scott M.,
Otte Kimberly D.
Publication year - 1989
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02535083
Subject(s) - desmosterol , chromatography , high performance liquid chromatography , chemistry , sterol , cholesterol , cholesterol oxidase , elution , methanol , biochemistry , organic chemistry
A simple and sensitive method to analyzed 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 C 18 reversed phase column (3 μm, 75×4.6 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 absoprtion at 240 nm. As little as 10 pmoles of sterol can be quantified under these conditions.