Fast liquid chromatographic assay of androgen aromatase activity.

A rapid, sensitive nonradiometric assay method for the enzymatic aromatization of androgens has been developed using reversed-phase fast liquid chromatography. The use of a 3-microns-particle, 5-cm-long column allowed analysis of androstenedione, estrone, testosterone, and estradiol within 1 min. The reliability of the method was demonstrated by measuring the aromatase activity of human placental microsomes and that of the purified reconstituted aromatase system using both substrates. The rapid analysis enabled the processing of a large number of samples in a short time, which makes the present method especially suitable for the analysis of chromatographic fractions obtained during enzyme purifications and for enzyme kinetic studies.