[en] A selective and sensitive ultra-performance liquid chromatography (UPLC)–electrospray ionization-tandem mass spectrometry (MS) method for simultaneous determination of bupropion and its main metabolites, hydroxybupropion, erythrohydrobupropion, and threohydrobupropion, in human whole blood is presented. The sample preparation consists of cleanup protein precipitation with methanol combined with a solid-phase extraction on Oasis HLB cartridges. Analytes were separated on a Waters Acquity UPLC® BEH phenyl column using a binary mobile phase consisting of ammonium formate buffer (2 mM, pH 4) and acetonitrile. Detection was performed on a Waters Acquity UPLC system coupled to a Quattro Premier triple-quadrupole MS in positive ion selected reaction monitoring. Internal standards were bupropion-d9 and hydroxybupropion-d6. Linearity was from 5 to 1000 ng/mL for bupropion and from 10 to 2000 ng/mL for metabolites. Accuracy profiles (80–120%), precision (< 15%), and limits of detection (1 ng/mL for bupropion and 2 ng/mL for metabolites) were also evaluated and responded to all criteria of validation. The aim of this study was to compare this presented method with a previously described method developed on a classic liquid chromatography–tandem MS system.
J.J. Steward, H.J. Berkel, R.C. Parish, M.R. Simar, A. Syed, J.A. Bocchini, J.T. Wilson, and J.E. Manno. Single-dose pharmacokinetics of bupropion in adolescents: effects of smoking status and gender. J. Clin. Pharmacol. 41: 770-778 (2001).
B.R. Cooper, C.M. Wang, R.F. Cox, R. Norton, V. Shea, and R.M. Ferris. Evidence that the acute behavioural and electrophysiological effects of bupropion (Wellbutrin®) are mediated by a noradrenergic mechanism. Neuropsychopharmacol. 11: 133- 141 (1994).
H.G. Kinnell. Bupropion for smokers: drug is almost identical in structure to diethylpropion, a controlled drug. Br. Med. J. 322: 431-432 (2001).
K.K Loboz, A.S. Gross, J. Ray, and A.J. McLachlan. HPLC assay for bupropion and its major metabolites in human plasma. J. Chro-matogr. B 823: 115-121 (2005).
R. Coles and E.D. Kharasch. Stereoselective analysis of bupropion and hydroxybupropion in human plasma and urine by LC/MS/MS. J. Chromatogr B 857: 67-75 (2007).
J. Posner, A. Bye, K. Dean, A.W. Peck, and P.D. Whiteman. The disposition of bupropion and its metabolites in healthy male volunteers after single and multiple doses. Eur. J. Clin. Pharmacol. 29: 97-103 (1985).
S.R. Faucette, R.L. Hawke, E.L. Lecluyse, S.S. Shord, B. Yan, R.M. Laethem, and C.M. Lindley. Validation of bupropion hydroxylation as a selective marker of human cytochrome P450 2B6 catalytic activity. Drug Metab. Dispos. 28: 1222-1230 (2000).
N.N. Bumpus, U.M. Kent, and P.F. Hollenberg. Metabolism of efavirenz and 8-hydroxyefavirenz by P450 2B6 leads to inactivation by two distinct mechanisms J. Pharmacol. Exp. Ther. 318: 345-351 (2006).
S. Palovaara, O. Pelkonen, J. Uusitalo, S. Lundgren, and K. Laine. Inhibition of cytochrome P450 2B6 activity by hormone replacement therapy and oral contraceptive as measured by bupropion hydroxylation. Clin. Pharmacol. Ther. 74: 326-333 (2003).
T.P. Rohrig and N.G. Ray. Tissue distribution of bupropion in a fatal overdose. J. Anal. Toxicol. 16: 343-345 (1992).
V. Ramcharitar, B.S. Levine, B.A. Goldberger, and Y.H. Caplan. Bupropion and alcohol fatal intoxication: case report. Forensic Sci. Int. 56: 151-156 (1992).
M. Mercerolle, R. Denooz, G. Lachatre, and C. Charlier. A fatal case of bupropion (Zyban) overdose. J. Anal. Toxicol. 32: 192-196 (2008).
Arlenda Home Page. E.noval Version V2.0e Prod http://www.arlenda.com (accessed September 2009).
FDA Home Page. Guidance for Industry: Bioanalytical Method Validation. http://www.fda.gov (accessed December 2009).