抄録
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.
