To investigate the metabolic pattern of such a drug having two counter functional groups in the molecule : a hydrophylic sulfate group and a hydrophobic aromatic ring, as 17-sulfate of 17β-hydroxyestra-1, 3, 5 (10)-triene (3-deoxyestradiol) was incubated with rat hepatic microsomes under a reduced nicotinamide adenine dinucleotide phosphate-generating system. The metabolites assigned were mono-and dihydroxylated products hydroxylated only at the aromatic ring. The phenolic products having hydroxyl group at the position 2, 3, or 4 were detected in the male and female rats. The two catechol products having hydroxyl groups at 2, 3 and 3, 4 were detected only in the male rats. No other metabolites were formed except a phenol product hydroxylated at the position 1 in the case of the male rats. These metabolisms were confirmed to proceed without removal of the sulfate group. A simple method for the determination of these metabolites was established by using reversed-phase high-performance liquid chromatography. The devised technique involves an elution with a mixture of 0.5% NH4H2PO4 (pH 3.0) and methanol on a TSK-Gel ODS-120A column with electrochemical and/or ultraviolet detectors.
