Simple determination of 3alpha-hydroxy fraction and 3beta-hydroxy fraction of urinary 17-ketosteroids.

Abstract

We describe a method for determination of the 3alpha-hydroxysteroid fraction and 3beta-hydroxysteroid fraction of 17-ketosteroids in urine. The method includes enzymatic hydrolysis with beta-glucuronidase and extraction with ethyl acetate and color development of 3beta-hydroxysteroids by 3beta-hydroxysteroid oxidase and of total 17-ketosteroids by the Zimmermann reaction. The value for 3alpha-hydroxysteroids is calculated by subtracting the value for 3beta-hydroxysteroids from the value for total 17-ketosteroids. Recovery was assessed and results compared with those by an accepted method. Results were satisfactory for measuring these fractions of 17-ketosteroids.