Development of a chromatographic bioreactor based on immobilized beta-glucuronidase on monolithic support for the determination of dextromethorphan and dextrorphan in human urine

Journal of Pharmaceutical and Biomedical Analysis
Enrica CalleriGabriele Caccialanza

Abstract

We here reported the development and application of an immobilized enzyme reactor (IMER) based on beta-glucuronidase to the on-line determination of urinary molar ratios of dextromethorphan (DOMe)/dextrorphan (DOH) for the assessment of the metabolic activity of CYP2D6, a genetically variable isoform of cytochrome P-450 (CYP). beta-Glucuronidase was immobilized on an HPLC monolithic aminopropyl silica support. Catalytic activity and stability of the chromatographic reactor were evaluated using 8-hydroxyquinoline glucuronide (8-HQG) as substrate. The IMER was coupled through a switching valve to a reversed-phase column (C8) for the simultaneous determination of dextromethorphan and its main metabolite dextrorphan. On purpose a selective reversed-phase ion pair HPLC method coupled with fluorescence detection was developed. Urine samples were first centrifuged to remove insoluble materials and then aliquots of the supernatants were injected into the coupled-column analyser. Linearity, precision and accuracy of the method were established. The method reliability was verified by comparing our data with previous data of a phenotyping study carried out by the Poison Control Centre of Pavia-Clinical Toxicology Division.

References

Oct 29, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·M P Di MarcoI W Wainer

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Citations

Mar 31, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Anna Maria Girelli, Enrico Mattei
Jun 6, 2014·Pakistan Journal of Biological Sciences : PJBS·Sekar RajanK Elango
Sep 15, 2006·Journal of Separation Science·Rangan Mallik, David S Hage
Apr 13, 2004·Nature Neuroscience·Kalanit Grill-SpectorNancy Kanwisher
Jul 13, 2006·Journal of Separation Science·John E SchielDavid S Hage

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