Application of overpressured layer chromatography combined with digital autoradiography and mass spectrometry in the study of deramciclane metabolism

Overpressured layer chromatography was combined with the highly sensitive a nd rapid digital autoradiography (DAR) and mass spectrometry to separate, d etect, and identify H-3- and C-14-labeled deramciclane metabolites in diffe rent biological matrixes. Several minor and major metabolites were separate d from plasma and urine samples. The radioactive metabolites localized by D AR were scraped from the thin-layer chromatographic plate and transferred t o a mass spectrometer for structure identification. Several metabolites wer e isolated and characterized, including hydroxy-N-desmethyl deramciclane, w hich is described in detail. The combination of techniques is efficient and has good sensitivity: about 2 mu g metabolite from a biological matrix was isolated and identified this way.