title: New method for assaying free and total cholesterol in cultured cells by high-pressure liquid chromatography.
abstract: We developed a simple, sensitive and accurate method for assaying cellular free and total cholesterol by monitoring 4-cholesten-3-one, a conversion product of the cholesterol oxidase-catalyzed oxidation of the free cholesterol that has a strong chromophoric alpha, beta-unsaturated ketone at 240 nm, using a high-pressure liquid chromatographic system. This method measured picomole quantities of free and total cholesterol and precisely determined their concentrations in cells (10(4) range) in culture using 7 beta-hydroxycholesterol as an internal standard.
authors.affiliation: First Department of Internal Medicine, Hiroshima University School of Medicine, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Kuga
authors.forename: Y
authors.initials: Y
authors.lastname: Okura
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
authors.forename: Y
authors.initials: Y
authors.lastname: Yasunobu
authors.forename: K
authors.initials: K
authors.lastname: Nomura
authors.forename: Y
authors.initials: Y
authors.lastname: Toyota
authors.forename: H
authors.initials: H
authors.lastname: Ohtani
authors.forename: T
authors.initials: T
authors.lastname: Shingu
authors.forename: G
authors.initials: G
authors.lastname: Kajiyama
mesh_terms: D015415:Biomarkers; D002478:Cells, Cultured; D002783:Cholestenones / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015415:Biomarkers; D002783:Cholestenones; C000475:cholest-4-en-3-one; D002784:Cholesterol