A Method for the Rapid Hydrolysis of Urinary Glucuronoside Metabolites of Testosterone.

..A.LMOST ALL the neutral urinary 17-ketosteroids are conjugated with either sulfuric or glucuronic acids. These conjugates can be cleaved by strong acids at elevated temperatures (1), or by solvolysis in either ethyl acetate or tetrahydrohiran, each containing perchioric acid (2). Selective hydrolysis of steroid-glucuronosicles is achieved with $-glucuromi_ idase, although in some 1i1#{149}illes, prolonged incubation periods are requirecl for complete hydrolysis. More rapid hydrolysis with $-glucuroiiidase has been reported wilen the urinary conjugates are extracted with an ether-alcohol solvent mixture prior to enzymatic treatment (3, 4). Ethyl acetate, another solvent that has beeli used to extract conjugates (5, 6), removes less nonsteroidal material from urine than the etheralcohol mixture (2). This is desirable if procedures for studying intact steroid-conjugates are planned, in addition to routine steroid determination after hydrolysis. This report evaluates $-glucuronidase hydrolysis of ethyl acetate-extracted conjugates. This technic has certain practical advantages over enzymatic treatment of urine.