Determination of 21-Hydroxycorticosteroids in Human Urine by High-Performance Liquid Chromatography with Fluorescence Detection

概要

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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 α-dicarbonyl compounds. The quinoxalines are separated within 70min on a reversed-phase column (TSK gel ODS-120T) by stepwise elution with mixtures of methanol, acetonitrile, and 1.0M ammonium acetate. The detection limits are 0.14-29.4pmol at a signal-to-noise ratio of 3 in a 50-μl injection volume. This sensitivity permits precise determination of hydrocortisone, cortisone, corticosterone, and their tetrahydro derivatives in 500 μl of normal human urine.
社団法人日本薬学会の論文
1989-11-25