Simultaneous determination of cholesterol and cholestanol in human serum by high-performance liquid chromatography using 3-(5,6-methylenedioxy-2-phthalimidyl)benzoyl azide as precolumn fluorescent labelling reagent.
A fluorescent labelling reagent, 3-(5,6-methylenedioxy-2-phthalimidyl) benzoyl azide, designed for the determination of alcohols by precolumn high-performance liquid chromatography, has been applied to the simultaneous determination of cholesterol and cholestanol in human serum. The reagent reacts with cholesterol and cholestanol at 140 degrees C for 10 min to produce the fluorescent derivatives, which can be separated on a reversed-phase column with acetonitrile-ethanol-water (60:35:7.5, v/v) as eluent. The detection limits for cholesterol and cholestanol were 45 and 50 fmol per injection (20 microliters), respectively. The values of cholesterol and cholestanol in normal human sera were 135-212 mg/dl and 137-928 micrograms/dl, respectively.
Tsuruta
Y
Y
Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Hiroshima, Japan.
Teranishi
T
T
Date
Y
Y
Kohashi
K
K
D001386:Azides / Q000737:chemistry*; D004083:Cholestanol / Q000097:blood*; D002784:Cholesterol / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006937:Hypercholesterolemia / Q000097:blood; D015394:Molecular Structure; D010797:Phthalimides / Q000737:chemistry*; D012016:Reference Values; D015203:Reproducibility of Results
D001386:Azides; D005456:Fluorescent Dyes; D010797:Phthalimides; C083701:3-(5,6-methylenedioxy-2-phthalimidyl)benzoyl azide; D004083:Cholestanol; D002784:Cholesterol