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Assessment of tandem mass spectrometry and high‐resolution mass spectrometry for the analysis of bupivacaine in plasma
Authors:Fleur Gaudette  Javier Benito  Paulo Steagall  Francis Beaudry
Affiliation:1. Centre de Recherche du Centre Hospitalier Universitaire de l'Université de Montréal, Montréal, Québec, Canada (CRCHUM);2. Département de Sciences Cliniques, Faculté de Médecine Vétérinaire, Université de Montréal, Saint‐Hyacinthe, Québec, Canada;3. Groupe de Recherche en Pharmacologie Animal du Québec, Département de Biomédecine Vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, Saint‐Hyacinthe, Québec, Canada
Abstract:Triple quadrupole mass spectrometers coupled with high performance liquid chromatography are workhorses in quantitative bioanalyses. They provide substantial benefits including reproducibility, sensitivity and selectivity for trace analysis. Selected reaction monitoring allows targeted assay development but datasets generated contain very limited information. Data mining and analysis of nontargeted high‐resolution mass spectrometry profiles of biological samples offer the opportunity to perform more exhaustive assessments, including quantitative and qualitative analysis. The objectives of this study were to test method precision and accuracy, to statistically compare bupivacaine drug concentration in real study samples and to verify if high‐resolution and accurate mass data collected in scan mode can actually permit retrospective data analysis, more specifically, extract metabolite related information. The precision and accuracy data presented using both instruments provided equivalent results. Overall, the accuracy ranged from 106.2 to 113.2% and the precision observed was from 1.0 to 3.7%. Statistical comparisons using a linear regression between both methods revealed a coefficient of determination (R2) of 0.9996 and a slope of 1.02, demonstrating a very strong correlation between the two methods. Individual sample comparison showed differences from ?4.5 to 1.6%, well within the accepted analytical error. Moreover, post‐acquisition extracted ion chromatograms at m/z 233.1648 ± 5 ppm (M ? 56) and m/z 305.2224 ± 5 ppm (M + 16) revealed the presence of desbutyl‐bupivacaine and three distinct hydroxylated bupivacaine metabolites. Post‐acquisition analysis allowed us to produce semi‐quantitative evaluations of the concentration–time profiles for bupicavaine metabolites. Copyright © 2015 John Wiley & Sons, Ltd.
Keywords:bupivacaine  liquid chromatography tandem mass spectrometry  bioanalysis  drug metabolism and pharmacokinetics  orbitrap mass spectrometer  high resolution  triple quadrupole mass spectrometer
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