Development and Validation of a Rapid Liquid Chromatography Tandem Mass Spectrometry Method for Simultaneous Quantification of Bupropion, Hydroxybupropion, Sertraline and Desmethylsertraline in Human Plasma

Abstract

Objectives: Bupropion and Sertraline are frequently used together in antidepressant treatment; therefore, analytic
methods with high sensitivity are needed to provide information for therapeutic drug monitoring (TDM) and drug-
drug interaction (DDI) studies. The present research work describes the development and validation of a Liquid
chromatography tandem mass spectrometry (LC-MS/MS) assay for simultaneous quantitation of Bupropion,
Hydroxybupropion, Sertraline, and Desmethylsertraline in human plasma. New Approach: Plasma samples were
prepared by protein precipitation with acetonitrile containing 0.1% formic acid. We used an acquity UPLC BEH
C18 column with gradient elution and positive electrospray ionization in multiple reaction monitoring mode
for chromatographic separation. Method validation was performed in accordance with the bioanalytical method
validation guidelines published by the U.S. Food and Drug Administration (FDA) and the European Medicines
Agency (EMA). Results: All analytes were separated within 6 min, and verification of linearity (r2 > 0.999)
was obtained for each compound with a lower limit of quantitation (defined as <30% relative standard deviation
ranging from 0.2 to 5 ng/mL. The lower limits of quantifications were 1 ng/mL for bupropion, 2 ng/mL for
hydroxybupropion, and 0.5 ng/mL for sertraline and desmethylsertraline, respectively. The matrix effects are very
low, and the carryover was negligible with over 94% recoveries. The accuracy was from 96.67% to 101.41%,
and the precision was all under 5%. All the conditions studied for stability of analyte were confirmed by the
two manufacturers. Conclusion: The validated LC-MS/MS method is specific, rapid, and sensitive, suitable for
pharmacokinetic, TDM, and antidepressant DDI studies