The conjugated steroids. IV. The hydrolysis of ketosteroid sulfates.

Specific hydrolysis of the separate groups of steroid conjugates appeared to offer the simplest technique for achieving the hydrolysis without chemical modification of the steroids (3-5). A partial solution to this problem was offered when it was found that steroid glucuronides could be hydrolyzed specifically by the use of /3-glucuronidase concentrates of either animal (Cohen (3)) or bacterial (Buehler et al. (4)) origin. Before the enzyme hydrolysis could be considered to be quantitative, it was necessary to account for the 40 to 50 per cent of the total 17-ketosteroids in urine that were not liberated by this hydrolysis. Phenolsulfatase preparations have been found to be effective for the hydrolysis of estrone sulfate (Cohen and Bates (6)). No one has as yet reported the successful use of any sulfatase, either of the phenol or alcohol type, for the hydrolysis of neutral steroid sulfates (Buehler et al. (7)). A number of non-enzymatic procedures have been suggested for the hydrolysis of steroid sulfates in urine. Most of these studies have involved the determination of the degree of hydrolysis of steroid sulfates without considering the possibility of chemical alterations effected during these procedures. Bitman and Cohen (2), using a modified Talbot et al. (8) hydrolysis in buffer, pointed out that apparently only the unsaturated P-steroid sulfates are hydrolyzed by this procedure. The isolation of about 8 per cent of the “total”’ ketosteroids from normal male urine as dehydro-