Liquid chromatography–tandem mass spectrometric assay for the mutated BRAF inhibitor dabrafenib in mouse plasma.

Abstract: A quantitative bioanalytical liquid chromatography–tandem mass spectrometric (LC–MS/MS) assay for the mutated BRAF inhibitor dabrafenib was developed and validated. Plasma samples were pre-treated using protein precipitation with acetonitrile containing PLX4720 as internal standard. The extract was directly injected into the reversed-phase chromatographic system after dilution with water. The eluate was transferred into the electrospray interface with positive ionization and the analyte was detected in the selected reaction monitoring mode of a triple quadrupole mass spectrometer. The assay was validated in a 2–2000ng/ml calibration range with r 2 =0.993±0.002 for linear regression with quadratic weighting (n =5). Within day precisions (n =6) were 3.3–5.2%, between day (3 days; n =18) precisions 4.7–8.2%. Accuracies were between 95–104% for the whole calibration range. The drug was sufficiently stable under all relevant analytical conditions. Finally, the assay was successfully used to determine drug pharmacokinetics in mice. [Copyright &y& Elsevier]