Stereoselective analysis of bupropion and hydroxybupropion in human plasma and urine by LC/MS/MS

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 857; no. 1; pp. 67 - 75
• Main Authors: Coles, Rebecka, Kharasch, Evan D.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.09.2007; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Antidepressive Agents, Second-Generation - blood; Antidepressive Agents, Second-Generation - chemistry; Antidepressive Agents, Second-Generation - urine; Biological and medical sciences; Bupropion; Bupropion - analogs & derivatives; Bupropion - blood; Bupropion - chemistry; Bupropion - pharmacokinetics; Bupropion - urine; Chiral analysis; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Humans; Hydroxybupropion; LC-MS-MS; Mass spectrometry; Medical sciences; Pharmacology. Drug treatments; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Solid Phase Extraction - instrumentation; Solid Phase Extraction - methods; Specimen Handling - instrumentation; Specimen Handling - methods; Spectrometry, Mass, Electrospray Ionization; Stereoisomerism; Tandem Mass Spectrometry; LC–MS–MS; Chiral analysis; Bupropion; Mass spectrometry; Hydroxybupropion; Human; Urine; Biological fluid; Psychotropic; Liquid chromatography; Blood plasma; Stereoselectivity; Optical resolution; Analysis; Antidepressant agent; LC-MS-MS
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2007.07.007

A sensitive, stereoselective assay using solid phase extraction and LC–MS–MS was developed and validated for the analysis of ( R)- and ( S)-bupropion and its major metabolite ( R, R)- and ( S, S)-hydroxybupropion in human plasma and urine. Plasma or glucuronidase-hydrolyzed urine was acidified, then extracted using a Waters Oasis MCX solid phase 96-well plate. HPLC separation used an α 1-acid glycoprotein column, a gradient mobile phase of methanol and aqueous ammonium formate, and analytes were detected by electrospray ionization and multiple reaction monitoring with an API 4000 Qtrap. The assay was linear in plasma from 0.5 to 200 ng/ml and 2.5 to 1000 ng/ml in each bupropion and hydroxybupropion enantiomer, respectively. The assay was linear in urine from 5 to 2000 ng/ml and 25 to 10,000 ng/ml in each bupropion and hydroxybupropion enantiomer, respectively. Intra- and inter-day accuracy was >98% and intra- and inter-day coefficients of variations were less than 10% for all analytes and concentrations. The assay was applied to a subject dosed with racemic bupropion. The predominant enantiomers in both urine and plasma were ( R)-bupropion and ( R, R)-hydroxybupropion. This is the first LC–MS/MS assay to analyze the enantiomers of both bupropion and hydroxybupropion in plasma and urine.