ENZYMATIC ESTIMATION OF STEROIDS IN HUMAN URINE1

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Abstract
The enzymatic microestimation of urinary steroid alcohols by means of ±- and ²-hydroxysteroid dehydrogenases is described. These highly purified enzymes catalyze selective and stereospecific oxidations of certain hydroxy-steroids to ketosteroids. Diphosphopyridine nucleotide functions as hydrogen acceptor and the assays are based on the spectrophotometric measurement of the reduced pyridine nucleotide. Methods are described for the preparation of extracts of human urine suitable for enzymatic assay. The reliability of the procedures has been established by a variety of criteria including the recovery of physiological amounts of steroids added to urineValues for the excretion of ±- and ²-hydroxysteroids have been determined in normal adult subjects of both sexes and in several patients with abnormalities of the endocrine system. The estimation of individual urinary steroids separated by paper chromatography is demonstrated. THE estimation of urinary steroids in man has been of interest for approximately 20 years since the introduction by Zimmermann (1) of a simple colorimetric test for 17-ketosteroids (17-KS), the determination of which has been widely practiced and clinically useful.