New method for assaying free and total cholesterol in cultured cells by high-pressure liquid chromatography.
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.
Hayashi
K
K
First Department of Internal Medicine, Hiroshima University School of Medicine, Japan.
Kuga
Y
Y
Okura
Y
Y
Tanaka
K
K
Yasunobu
Y
Y
Nomura
K
K
Toyota
Y
Y
Ohtani
H
H
Shingu
T
T
Kajiyama
G
G
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
D015415:Biomarkers; D002783:Cholestenones; C000475:cholest-4-en-3-one; D002784:Cholesterol