High-resolution biomedical gas chromatography : Determination of human urinary steroid metabolites using glass open tubular capillary columns

Abstract

Recently developed analytical procedures for the qualitative and quantitative analysis of human urine for major and minor steroid metabolites are described. Steroid profile samples were obtained by enzymic hydrolysis with β-glucuronidase and sulfatase. Methoxime-trimethylsilyl ether derivatives of steroid metabolites were prepared; the recommended procedure converts all ketone groups (except the 11-one group) into methoxime groups and all hydroxyl groups into trimethylsilyl ether groups. These derivatives are thermally stable, readily volatilized, not subject to dehydration or adsorption on gas chromatographic columns, and suitable for both qualitative and quantitative analytical studies. Thermostable glass open tubular capillary columns, coated with the non-polar phase SE-30, and containing dispersed particles of silanized silicic acid, were used for the gas chromatographic separation. Illustrations of profiles for normal female and male subjects, and patients with a testosterone-secreting ovarian tumor, congenital adrenal insufficiency and a dehydroepiandrosterone-secreting adrenal tumor are included.