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Journal of Clinical Endocrinology & Metabolism Vol. 22, No. 12 1234-1241
doi:10.1210/jcem-22-12-1234
Copyright © 1962 by the Endocrine Society.
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A Study of Methods for the Determination of Total, Grouped and Individual Urinary 17-Ketosteroids

JOSEPH W. GOLDZIEHER, M.D., LEONARD R. AXELROD, PH.D.,, technical assistance, Eva C. Riha and Christina M. Tobias

Department of Endocrinology Southwest Foundation for Research and Education San Antonio, Texas
Department of Biochemistry, Southwest Foundation for Research and Education San Antonio, Texas

The 17-ketosteroid content of urine has been examined by 4 methods: a clinical "total 17-ketosteroid" determination after acid hydrolysis; a total ketosteroid determination on the ketonic fraction from a micro-Girard procedure; a simple column chromatographic separation of the C19O2 and C19O3 steroids of an extract of urine subjected to enzyme hydrolysis and solvolysis; and a 2-stage paper chromatographic separation of androsterone, etiocholanolone, dehydroepiandrosterone and the C19O3 steroids of a similar urine extract. Paper chromatographic analysis revealed that up to 90 per cent of the clinical "total 17-ketosteroid" value may consist of nonspecific chromogens, thus largely invalidating the quantitative significance of such measurements. Girard fractionation achieved satisfactory purification with some loss of steroids. The column chromatographic method proved more practical, yielding 2 biologically significant ketosteroid fractions which correlated well with the results of paper chromatography.

This study was supported by Grant A-3269, National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, USPHS.

Received April 6, 1962.






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Copyright © 1962 by The Endocrine Society