PAPIERCHROMATOGRAPHIE DER 17-KETOSTEROIDE MIT QUANTITATIVER AUSWERTUNG

Abstract

ABSTRACT The method presented is of practical value in the study of the excretion of the 17-ketosteroids, since it permits after hydrolysis with hydrochloric acid and following purification quantitative determination of: I: 11-oxygenated 17-ketosteroids II: 3β-hydroxy-androst-5-en-17-one (DHA) III: 3β-hydroxy-androstan-17-one IV: 3α-hydroxy-5β-androstan-17-one (aetiocholanolone) V: 3α-hydroxy-5α-androstan-17-one (androsterone) VI: androstane-3,17-dione VII: androst-5-ene-3β-chloro-17-one with other artifacts. The separation on paperchromatogram in 7 zones is achieved by running the mobile phase two times using dimethylformamide/methylcyclohexane as solvent system. The Zimmermann reaction is employed to reveal the zones marked by reference standards which were run in lanes parallel to the extracts being tested. Then the zones are cut out and extracted with decomposition of the dyestuff and quantitative regeneration of the original ketosteroids. Pigments in the extracts were corrected for by measuring the extinctions developed in the absence of dinitrobenzene. The ketosteroids are estimated photometrically by the Zimmermann reaction. After discussing the results of recovery and related experiments, values are given concerning fractional analysis of urinary 17-ketosteroids in two normal women, and compared with tabulated data from literature. The recommended fractionation is especially valuable in the investigation of endocrinopathies, data on this will be published in a subsequent paper.