In vitro metabolism of estrone-4-14C and estrone-6,7-3H-3-sulfate by laying hen liver homogenates.

Abstract

Estrone-4-14C and estrone-6,7-3H-3-sulfate were simultaneously incubated with hen liver homogenates prepared from fowl in the laying stage. Purification and identification of conjugated metabolites was made by two different methods: (a) an indirect procedure involving hydrolysis with a sulfatase preparation, followed by ether extraction, Girard separation, Celite partition chromatography, and crystallization to constant specific activity of the resulting free estrogens; (b) a direct identification of estrogen sulfates by paper chromatography in three different solvent systems, again followed by recrystallization. The unconjugated (free) incubation products were identified by the former of the two methods.Liver homogenates from the fowl were capable of catalyzing a rapid reduction of estrone to estradiol-17β and of estrone-3-sulfate to estradiol-17β-3-sulfate, the latter apparently without removal of the sulfate group. Under the conditions of the experiments, a slower dehydrogenation of estradiol-17β to estrone and of estradiol-17β-3-sulfate to estrone-3-sulfate was also indicated.