DESTRUCTION OF 3-&HYDROXY 17-KETOSTEROIDS DURING HYDROLYSIS WITH HYDROCHLORIC ACID* BY JOEL BITMANt

Acid hydrolysis has long been used to free the urinary 17-ketosteroids from the water-soluble combinations in which they are excreted. Both hydrochloric acid (13,5,11) and sulfuric acid (15)’ have been employed, for this purpose. The acid treatment, while effecting relatively complete hydrolysis, may also result in destruction (13, 5, 6,3, IS), chlorine substitution (2), dehydration (20,4, 12), and in stereoisomeric transformations (6, 16). Attempts to obviate the destructive action of acid hydrolysis have been made by Talbot and his associates. They described an enzymatic hydrolytic procedure (18) and also a method of hydrolysis utilizing barium chloride at pH 5.8 (19). Their studies were conducted on urine from patients with adrenal cortical carcinoma and adrenal cortical hyperplasia (18). We have investigated the BaClz hydrolysis of the conjugated 17ketosteroids in specimens of normal male urine and have obtained by this method a much higher P (C, hydroxyl cis to the Cl0 methyl) fraction than is usually reported for normal urine. The experiments reported below further indicate that a large proportion of the p-17-ketosteroids is changed to an “a” fraction (no cis Ct hydroxyl group) by the usual HCl hydrolytic technique.