The extraction and hydrolysis of steroid monoglucuronides.

1. A method in use for the extraction of urinary steroid conjugates has been applied to study the recovery of synthetic steroid monoglucuronides from aqueous solution. 2. In the presence of dissolved ammonium sulphate (50g./100ml.), ether-ethanol (3:1, v/v, 3x0.5vol.) extracted the monoglucuronides of steroids of the C(18), C(19) and C(21) series, quantitatively at values pH2-9. 3. The hydrolysis of the synthetic steroid monoglucuronides by beta-glucuronidase (Patella vulgata) has been examined with reference to the pH value of the medium, enzyme concentration and substrate concentration. 4. The rate of hydrolysis of steroid monoglucuronides was dependent upon steroid structure and upon site of conjugation. 5. The rate of hydrolysis of the monoglucuronides decreased in the order C-3 (phenolic) >C-3beta>C-17beta>C-3alpha.