Corticosteroid metabolism in isolated rat kidney in vitro. III. Structure analysis of lipid soluble metabolites of corticosterone.

We have previously demonstrated that isolated rat kidneys in vitro convert corticosterone (B). Four lipid soluble metabolites (met I, II, III and IV) have been identified which differ in polarity from the parent hormone [2, 5, 6, 11, 15, 16]. In the present experiments these metabolites have been extracted from perfusate after 4 h of recirculation through isolated kidneys of male and female rats. Subsequently they have been separated by HPLC using a polar stationary phase system and n-hexane and isopropanol as eluents. The chromatographic comparison of met II with authentic 20 alpha- and 20 beta-isomers documented that met II is identical with 20 beta-dihydro-B. Measurements of the mass spectra of the purified samples revealed the following structures: met I = 20 beta-dihydro-11-dehydro-B, met II = 20 beta-dihydro-B, met III = 5 alpha-H-4,5-dihydro-B and met IV = 11-dehydro-B.