Metabolite analysis, isolation and purity assessment using various liquid chromatographic techniques combined with radioactivity detection.

During the discovery of metabolic routes of a drug candidate, radioactively labeled substances are administered. This study reports the multidimensional application of overpressured layer chromatography (OPLC) and high-performance liquid chromatography (HPLC) coupled with online or off-line nondestructive radioactivity detection methods in metabolism studies. Among these methods, digital autoradiography and flow-cell radioactivity detectors (RD) using solid scintillators are used. In this study, the hyphenation of OPLC with RD is reported. The application of the OPLC-RD technique is demonstrated on a metabolism study as well as the multidimensional chromatographic selectivity using normal-phase OPLC for the separation in the first dimension, followed by reversed-phase HPLC-RD, which provides additional selectivity to the separation. Information regarding the identity of radiolabeled metabolites and data obtained from spectroscopic methods could be advantageously used during structure elucidation.

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 and rapid digital autoradiography (DAR) and mass spectrometry to separate, detect, and identify 3H- and 14C-labeled deramciclane metabolites in different biological matrixes. Several minor and major metabolites were separated from plasma and urine samples. The radioactive metabolites localized by DAR were scraped from the thin-layer chromatographic plate and transferred to a mass spectrometer for structure identification. Several metabolites were isolated and characterized, including hydroxy-N-desmethyl deramciclane, which is described in detail. The combination of techniques is efficient and has good sensitivity: about 2 micrograms metabolite from a biological matrix was isolated and identified this way.

Planar chromatographic techniques in biomedicine: current status.

In planar chromatography (PLC), the solvent flows through a layer either by means of capillary forces [conventional thin-layer chromatography (TLC)] or by a forced-flow system (over-pressured layer chromatography). Phases and instrumentation currently available are briefly examined. The main applications in biomedicine are reviewed. Although silica gel TLC plates still predominate, interest in other phases is increasing. Unique detection features such as immunostaining are emphasized. Although gas chromatography and high-performance liquid chromatography have superseded TLC in the analysis of carbohydrates, amino acids and indole derivatives, interest in PLC continues to be high in lipid analysis.

Separation, quantitation and isolation of cannabinoids from Cannabis sativa L. by overpressured layer chromatography.

Two overpressured layer chromatography (OPLC) methods have been developed for the separation of neutral and acidic cannabinoids. The first is an adaptation of Korte's well known method to the OPLC system, which improves its reproducibility. The second one is a new technique based on the phenomenon of chromatographic solvent demixing. The eluent itself is also divided into zones. In the alpha-zone the neutral cannabinoids and in the beta-zone the acidic ones are separated. As a result of the good and reproducible separation, there is a possibility to quantitate cannabinoids by densitometry. The on-line version of OPLC proved suitable for the isolation of hemp constituents.