Quantification of the O- and N-demethylated metabolites of hydrocodone and oxycodone in human liver microsomes using liquid chromatography with ultraviolet absorbance detection
Date
2003
Authors
Menelaou, A.
Hutchinson, M.
Quinn, I.
Christensen, A.
Somogyi, A.
Editors
Advisors
Journal Title
Journal ISSN
Volume Title
Type:
Journal article
Citation
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 2003; 785(1):81-88
Statement of Responsibility
Andrew Menelaou, Mark R. Hutchinson, Ingvild Quinn, Anders Christensen and Andrew A. Somogyi
Conference Name
Abstract
High-performance liquid chromatographic assays for the O- and N-demethylated oxidative metabolites of hydrocodone and oxycodone formed in human liver microsomes are described. A solvent–solvent extraction/re-extraction procedure followed by reversed-phase HPLC with UV detection at 210 nm allows for the quantification of hydromorphone, norhydrocodone, oxymorphone and noroxycodone. Calibration curve concentration ranges were 0.63–400 μM (0.18–114 μg/ml) and 1.25–400 μM (0.36–114 μg/ml) for hydromorphone and norhydrocodone, respectively and 0.13–20 μM (0.04–6.03 μg/ml) and 1–200 μM (0.30–60 μg/ml) for oxymorphone and noroxycodone, respectively. Assay performance was determined by intra- and inter-assay precision and inaccuracies for quality control samples and was <15% for all metabolites at each quality control concentration. These methods provide good precision, accuracy and sensitivity for use in in vitro kinetic studies investigating the oxidative metabolism of hydrocodone and oxycodone in human liver microsomes.