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

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 2–2000 ng/ml calibration range with r2 = 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.

► The first validated bioanalytical assay for dabrafenib has been reported.
► The assay has successfully been validated in the 2–2000 ng/ml range.
► The drug is sufficiently stable under all conditions relevant for the assay.
► First pharmacokinetics in male wild type FVB mice have reported.