Enantioselective high-performance liquid chromatographic method for the determination of methadone and its main metabolite in urine using an AGP and a C8 column coupled serially |
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| Institution: | 1. Department of Clinical Biochemistry Bispebjerg Hospital, Bispebjerg Bakke, DK-2400 Copenhagen NV, Denmark;2. The Multidisciplinary Pain Clinic, Bispebjerg Hospital, Bispebjerg Bakke, DK-2400 Copenhagen NV, Denmark;3. Department of Bioanalytical Development, H. Lundbeck, Ottiliavej 9, DK-2500 Valby, Denmark;1. INSERM UMR 1027, 37 allées Jules Guesde, Toulouse F-31073, France;2. University of Toulouse III, 37 allées Jules Guesde, Toulouse F-31073, France;3. Department of Epidemiology and Public Health, CHU Toulouse, 37 allées Jules Guesde, Toulouse F-31073, France;4. Nutrition Unit, Department of Endocrinology, CHU Toulouse, 24 chemin de Pouvourville, Toulouse F-31059, France;1. Unit of Gynecological Oncology, Institute Clinic of Gynecology, Obstetrics and Neonatology (ICGON), Hospital Clinic of Barcelona, Universitat de Barcelona, IDIBAPS, 08036 Barcelona, Spain;2. Department of Pathology, Vall d''Hebron University Hospital, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain;3. Department of Gynecology, Vall d''Hebron University Hospital, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain;4. Department of Anesthesiology, Vall d''Hebron University Hospital, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain;5. Biomedical Research Group in Gynecology, Vall d’Hebron Research Institute, Universitat Autònoma de Barcelona, CIBERONC, 08035 Barcelona, Spain;1. University of Western Sydney (UWS), Australian Centre for Research on Separation Science (ACROSS), School of Science and Health, Locked Bag 1797, Penrith, NSW 2751, Australia;2. University of Western Sydney (UWS), Molecular Medicine Research Group (MMRG), School of Science and Health, Locked Bag 1797, Penrith, NSW 2751, Australia;1. Impact strategic research centre, school of medicine, Deakin university, PO Box 281, Geelong, Victoria 3220, Australia;2. Department of statistics, data science and epidemiology, Swinburne university of technology, Swinburne, Australia;3. Melbourne clinical school-western campus, the university of Melbourne, Saint-Albans, VIC, Australia;4. Department of epidemiology and preventive medicine, Monash university, Melbourne, VIC, Australia;5. University hospital of Geelong, Geelong, VIC, Australia;6. Department of psychiatry, the university of Melbourne, Parkville, VIC, Australia;7. Florey institute of neuroscience and mental health, Parkville, VIC, Australia;8. Orygen, the National centre of excellence in youth mental health, Parkville, VIC, Australia;9. Centre for adolescent health, Murdoch children''s research institute, Melbourne, Australia;10. Black Dog institute, Sydney, Australia |
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| Abstract: | A simple and sensitive method for the enantioselective high-performance liquid chromatographic determination of methadone and its main metabolite, EDDP, in human urine is described. (?)-(R)-Methadone, (+)-(S)-methadone, (+)-(R)-EDDP, (?)-(S)-EDDP and imipramine as an internal standard are detected by ultraviolet detection at 200 nm. The enantiomers of methadone and EDDP were extracted from human urine by a simple liquid–liquid extraction procedure. The extracted sample was reconstructed in mobile phase and the enantiomers of methadone and EDDP were quantitatively separated by HPLC on a short analytical LiChrospher RP8 column coupled in series with a chiral AGP column. Determination of all four enantiomers was possible in the range of 0.03 to 2.5 μM. The recoveries of methadone enantiomers and EDDP enantiomers added to human urine were about 90% and 80%, respectively. The method was applicable for determination of methadone enantiomers and the enantiomers of its main metabolite in urine samples from methadone maintenance patients and patients suffering from severe chronic pain. |
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