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N. B. TALBOT, R. A. BERMAN, E. A. MACLACHLAN, J. K. WOLFE, The Colorimetric Determination of Neutral Steroids (Hormones) in a 24-Hour Sample of Human Urine (Pregnanediol; Total, Alpha and Beta Alcoholic, and Non-Alcoholic 17- Ketosteroids), The Journal of Clinical Endocrinology & Metabolism, Volume 1, Issue 8, 1 August 1941, Pages 668–673, https://doi.org/10.1210/jcem-1-8-668
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Abstract
IT is becoming increasingly apparent that measurements of the urinary steroids are of value in clinical diagnosis and in studies of human endocrine physiology. Relatively simple procedures for the determination of the total, alpha and beta alcoholic, and non-alcoholic neutral 17-ketosteroids in a single twenty-four-hour urine sample have been reported (1, 2). Astwood and Jones (3) have recently perfected a simple procedure for the gravimetric determination of pregnanediol. The present paper reports a method whereby both the 17-ketosteroids and the pregnanediol may be determined colorimetrically on a single twenty-four-hour urine specimen.
The method involves (a) the simultaneous hydrolysis and extraction of steroids from urine with carbon tetrachloride, (b) separation of the neutral steroids into ketonic and non-ketonic fractions with Girard’s Reagent “T,” (c) colorimetric determination of pregnanediol in the non-ketonic fraction after purification according to Astwood and Jones and (d) colorimetric determination of the 17-ketosteroids in the ketonic fraction as described elsewhere (4).