Development of a linear dual column HPLC–MS/MS method and clinical genetic evaluation for tramadol and its phase I and II metabolites in oral fluid
- 194 Downloads
Tramadol is a centrally acting synthetic opioid analgesic and has received special attention due to its abuse potential and unexpected responses induced by CYP2D6 polymorphism. Oral fluid is an advantageous biofluid for drug analysis due to non-invasive sampling and high correlation of drug concentrations with plasma. However, few studies have been performed on distribution of tramadol and its metabolites in oral fluid. In the present study, a linear dual column HPLC–MS/MS method was developed and fully validated for the simultaneous determination of tramadol and its phase I [O-desmethyltramadol (ODMT), N-desmethyltramadol (NDMT) and N,O-didesmethyltramadol (NODMT)] and II metabolites in oral fluid. Furthermore, the distribution of tramadol and its metabolites, in relation to CYP2D6 genetic variations, in oral fluid was investigated following a clinical study including 23 subjects with CYP2D6*wt/*wt, CYP2D6*10/*10 or CYP2D6*5/*5. The validation results of selectivity, matrix effect, linearity, precision and accuracy were satisfactory. Pharmacokinetic parameters, such as Css,max and AUC0–τ of tramadol, NDMT and NODMT, in the CYP2D6*10/*10 group were significantly higher than those in the CYP2D6*wt/*wt group. Moreover, the ratios of ODMT/tramadol, NDMT/tramadol and NODMT/NDMT correlated well with the CYP2D6 genotypes. We demonstrated that oral fluid is a promising biofluid for pharmacokinetic evaluation in relation to genetic variations.
KeywordsTramadol Oral fluid Dual column HPLC–MS/MS CYP2D6 Genetic variation
This research was supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2015M3A9E1028327) and by the Basic Science Research Program of NRF funded by the Ministry of Education (NRF-2016R1A6A1A03011325).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Abdel-Rahman SM, Leeder JS, Wilson JT, Gaedigk A, Gotschall RR, Medve R, Liao S, Spielberg SP, Kearns GL (2002) Concordance between tramadol and dextromethorphan parent/metabolite ratios: the influence of CYP2D6 and non-CYP2D6 pathways on biotransformation. J Clin Pharmacol 42:24–29CrossRefPubMedGoogle Scholar
- Lee YJ, Choi JH, Song SH, Seo CH, Kim DS, Park IS, Choi KH, Na HK, Chung SJ, Lee MH, Shim CK (1998) Development of K-BE test, a computer program for the analysis of bioequivalence. J Kor Pharm Sci 28:223–229Google Scholar
- Mehvar R, Elliott K, Parasrampuria R, Eradiri O (2007) Stereospecific high-performance liquid chromatographic analysis of tramadol and its O-demethylated (M1) and N, O-demethylated (M5) metabolites in human plasma. J Chromatogr B Analyt Technol Biomed Life Sci. 852:152–159CrossRefPubMedGoogle Scholar
- Nobilis M, Kopecky J, Kvetina J, Chladek J, Svoboda Z, Vorisek V, Perlík F, Pour M, Kunes J (2002) High-performance liquid chromatographic determination of tramadol and its O-desmethylated metabolite in blood plasma. Application to a bioequivalence study in humans. J Chromatogr A 949:11–22CrossRefPubMedGoogle Scholar
- Peters FT (2006) Method validation. In: Polettini A (ed) Applications of LC–MS in toxicology. Pharmaceutical Press, London, pp 71–95Google Scholar
- Peters FT, Hartung M, Herbold M, Schmitt G, Daldrup T, Mußhoff F (2009) Appendix B, To the GTFCh Guidelines for quality assurance in forensic-toxicological analyses, requirements for the validation of analytical methods. https://www.gtfch.org/cms/images/stories/files/. Accessed 11 Nov 2017
- Tanaka H, Naito T, Mino Y, Kawakami J (2016) Validated determination method of tramadol and its desmethylates in human plasma using an isocratic LC-MS/MS and its clinical application to patients with cancer pain or non-cancer pain. J Pharm Health Care Sci 2:25CrossRefPubMedPubMedCentralGoogle Scholar
- Vazzana M, Andreani T, Fangueiro J, Faggio C, Silva C, Santini A, Garcia ML, Silva AM, Souto EB (2015) Tramadol hydrochloride: pharmacokinetics, pharmacodynamics, adverse side effects, co-administration of drugs and new drug delivery systems. Biomed Pharmacother 70:234–238CrossRefPubMedGoogle Scholar