Abstract
After incubating 18-hydroxydeoxycorticosterone (18-OH-DOC) with cytochrome P-450 11β in the reconstituted system, the products were analyzed with HPLC. There appeared two product-peaks on the chromatogram, one of which was identified as a peak of 18-hydroxycorticosterone (18-OH-B), an expected product of the 11β-hydroxylation. Another peak did not coincide with those of any known corticoids. This unidentified product was further purified, and the purified material was analyzed by gas chromatography-mass spectrometry ( GC MS ). The mass spectrum showed that the unidentified product is one of the structural isomers of 18-OH-B. A further analysis with 1H-NMR spectrometry indicated that a proton resonance peak of 19-CH 3 in 18-OH-DOC disappeared in the product and the methyl group of the substrate seemed to be converted to -CH 2OH. These results suggested that the unidentified product generated from 18-OH-DOC by P-450 11β-linked hydroxylase system may be 18,19-dihydroxydeoxycorticosterone (18,19,21-trihydroxypregn-4-ene-3,20-dione; 18,19-diOH-DOC), a hitherto unreported corticoid.
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