The hydrolysis of conjugated neutral steroids in urine.

Complete hydrolysis of known conjugates of neutral steroids in urine requires a combination of solvolysis and enzymatic hydrolysis. Procedures are established for optimal yield in the enzymatic part of the hydrolysis using different preparations of beta-glucuronidase. The effect of temperature and the use of conjugate extraction in the hydrolytic procedure is explored. It is stressed that relatively high concentrations of enzyme are necessary to insure full hydrolysis of a few conjugates that are particularly difficult to split by enzymatic hydrolysis.