Formation of 6 beta-OH-deoxycorticosterone from deoxycorticosterone by A6 cells

We have investigated the metabolism of deoxycorticosterone in A6 cells, a continuous cell line derived from the kidney of Xenopus laevis. A6 cells at confluence were incubated with serum-free culture media containing 2.5 microM [3H] deoxycorticosterone. When radioactive compounds in incubation media were separated by reverse-phase high-pressure liquid chromatography, the formation of polar metabolites was observed. One component of polar metabolites cochromatographed with 6 beta-hydroxydeoxycorticosterone. In order to identify this component more rigorously, large scale cultures were performed and this compound was separated and purified by reverse-phase high-pressure liquid chromatography. The purified material was analyzed by gas chromatography-mass spectrometry, secondary ion mass spectrometry, and nuclear magnetic resonance spectroscopy. These procedures revealed that this material was 6 beta-hydroxydeoxycorticosterone.