title: Determination of 21-hydroxycorticosteroids in human urine by high-performance liquid chromatography with fluorescence detection.
abstract: A simple and sensitive high-performance liquid chromatographic method with fluorescence detection for the determination of nineteen 21-hydroxycorticosteroids is described. The corticosteroids are oxidized by cupric acetate to form the corresponding glyoxal derivatives. The derivatives are converted into fluorescent quinoxalines by reaction with 1,2-diamino-4,5-methylenedioxybenzene, a fluorogenic reagent for alpha-dicarbonyl compounds. The quinoxalines are separated within 70 min on a reversed-phase column (TSK gel ODS-120T) by stepwise elution with mixtures of methanol, acetonitrile, and 1.0 M ammonium acetate. The detection limits are 0.14-29.4 pmol at a signal-to-noise ratio of 3 in a 50-microliter injection volume. This sensitivity permits precise determination of hydrocortisone, cortisone, corticosterone, and their tetrahydro derivatives in 500 microliters of normal human urine.
authors.forename: M
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: T
authors.initials: T
authors.lastname: Yoshitake
authors.forename: J
authors.initials: J
authors.lastname: Ishida
authors.forename: M
authors.initials: M
authors.lastname: Nakamura
mesh_terms: D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006889:Hydroxycorticosteroids / Q000652:urine*; D013050:Spectrometry, Fluorescence
chemical_list: D006889:Hydroxycorticosteroids