CHEMICAL METHOD FOR THE DETERMINATION OF 17-HYDROXYCORTICOSTEROIDS AND 17-KETOSTEROIDS IN URINE FOLLOWING HYDROLYSIS WITH β-GLUCURONIDASE*

Abstract

METHODS for the estimation of metabolites of the adrenocortical steroids thus far published have given only partially satisfactory estimates of adrenocortical function. A method has been developed which gives reproducible determinations of 17,21-dihydroxy-20-ketones and 17-ketosteroids present in urine, either in the free form or conjugated as glucuronides. The acid hydrolysis of urinary adrenal steroid metabolites has been accomplished with varying degrees of success by a number of investigators (1–4), but there is considerable evidence suggesting loss or alteration of steroids during the hydrolytic process (5, 6). It has been shown that a large part of the adrenocortical steroids in urine are conjugated as glucuronides (7–10). In the present method hydrolysis with β-glucuronidase was employed and consistent levels of adrenocortical steroid metabolites were found after chromatography on a Florisil column.