Identification of pregnane-3 (alpha), 17-diol-20-one as the steroid moiety of a new glucuronide isolated from human urine.

In 1943 one of us (H. S. S.) and coworkers (1) reported the isolation of a new steroid glucuronide from the urine of a young female pseudohermaphrodite.1 The free steroid was obtained by acid hydrolysis and was characterized tentatively as a steroid having an ac-OH at C-3 and a ketone group at C-20. Some further work was done on the steroid moiety in the laboratories of the School of Medicine, University of Pittsburgh, and Magee Hospital. Changing circumstances made it necessary for the original investigators to discontinue their study, and work on the identity of the steroid moiety was undertaken at the Mayo Foundation. This report covers the work done since the preliminary report. Urine was processed by Yenning’s (2) method for the isolation of sodium pregnanediol glucuronidate (NaPG). The glucuronidate obtained, however, differed from NaPG in physical appearance and solubility. Analysis indicated the formula C27H4i09Na, the sodium glucuronidate of a steroid with the formula C2iH3403. Reduction of Benedict’s reagent after hydrolysis and a positive result of Tollens’ naphthoresorcinol test indicated the presence of glucuronic acid. Several preparations were hydrolyzed for isolation of the steroid moiety. In the earlier work only material that had been extensively recrystallized was used. An acetate was prepared by Dr. K. Hofmann from some of the ketonic steroid so obtained, which was shown to be identical with the acetate obtained later. In the later work a relatively crude preparation of glucuronidate was used, and treatment of the steroid moiety with Girard’s Reagent T gave some pregnanediol in addition to the ketone previously described. The analytical results suggest that the highly purified