Determination of 17-oxosteroids in serum and urine by fluorescence high-performance liquid chromatography using dansyl hydrazine as a pre-labeling reagent

doi:10.1016/S0378-4347(00)84200-0

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 226, Issue 1, 13 November 1981, Pages 1-12

https://doi.org/10.1016/S0378-4347(00)84200-0 Get rights and content

Abstract

A fluorescence high-performance liquid chromatographic method is described for the determination of 17-oxosteroids in biological fluids. 17-Oxosteroids in urine samples are extracted with dichloromethane after enzymatic hydrolysis (β-glucuronidase—sulfatase), and dehydroepiandrosterone sulfate in serum samples is solvolysed with sulfuric acid in ethyl acetate. 17-Oxosteroids are labeled with dansyl hydrazine in trichloroacetic acid—benzene solution, and then chromatographed on the microparticulate silica gel column using dichloromethane—ethanol—water (400:1:2) as the mobile phase. The eluate is monitored by a fluorophotometer at 365 nm (excitation) and 505 nm (emission). Linearities of the fluorescence intensities (peak heights) with the amounts of various 17-oxosteroids were obtained between 60 and 1000 pg. The assay proved satisfactory with respect to sensitivity, precision and accuracy. The results obtained by a radioimmunoassay and this method were in good agreement (r = 0.964, n = 81) for serum dehydroepiandrosterone sulfate. This method is also useful for the simultaneous determination of individual 17-oxosteroids in serum and urine.

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Determination of 17-oxosteroids in serum and urine by fluorescence high-performance liquid chromatography using dansyl hydrazine as a pre-labeling reagent

Abstract

J. Chromatogr.

J. Chromatogr.

J. Chromatogr.

J. Chromatogr.

Steroids

Folia Endocrinol. Jap.

J. Clin. Endocrinol.