A method for the quantitative fractionation of urinary 17-oxo steroids with some observations on steroid excretion during administration of ACTH and in the adrenogenital syndrome

Abstract

A method is described for the analysis of the major urinary 17-ketosteroids. An extract containing the urinary 17-ketasteroid conjugates is hydrolyzed first with [beta]-glucuronidase, then by continuous extraction with ether at pH 0.8. The liberated 17-ketosteroids are purified by application of the Girard treatment and the ketonic fraction is treated with KHCO3 in aqueous methanol to hydrolyze any acetates formed in the Girard separation. The product is then separated into 2 fractions by chromatography on a column of alumina. The first fraction diluted contains the 17-ketosteroids with a C(11) and the 2d fraction contains the 17-ketosteroids with a C(11) O2 function. These 2 fractions are resolved into their component 17-ketosteroids by paper chromatography, the final estimation being made by applying the Zimmermann reaction to residues obtained after eluting the individual steroids from the paper. Recovery of added 17-ketosteroid through the complete procedure is 70-80%. Evidence is presented to show the effectiveness of the hydrolytic procedures. Normal values for 6 females and 2 males are given, and also values after stimulation with adrenocorticotrophic hormone, and for one case of the adrenogenital syndrome.