HYDROLYSIS OF URINARY NEUTRAL 17-KETOSTEROID CONJUGATES

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Abstract
Urine is hydrolyzed twice[long dash]1st at 100C for 18 hrs. at pH 4-10 (not critical); then after adding 0.1 volume 70% H2SO4, at lOO''C for 30 min. Ether extractions are performed after each hydrolysis. The total 17-keto steroid (17-KS) yield by this method is 119-174% higher than by standard hydrolysis methods. The 1st hydrolysis releases all steroids giving the dehydroisoandrosterone reaction of Jensen (Acta Endocrinol. 4: 140 and 374, 1950), and includes chiefly this steroid (about 80% of total 17-KS released in this hydrolysis). Probably most of the esters hydrolyzed in the 1st procedure are sulfates. The 2d hydrolysis affects chiefly glucuronides, and accounts for about 50-80% of the total 17-KS. No appreciable amts. of dehydroisoandrosterone-like material appears in this hydrolysis. The authors suggest that partition of 17-KS into dehydroisoandrosterone, total easily-hydrolyzable 17-KS, and acid-hydrolyzable 17-KS may increase the diagnostic value of the urinary 17-KS detn.