Liquid chromatography-tandem mass spectrometric assay for the tyrosine kinase inhibitor afatinib in mouse plasma using salting-out liquid-liquid extraction.

摘要: A quantitative bioanalytical liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay for afatinib, an irreversible inhibitor of the ErbB (erythroblastic leukemia viral oncogene homolog) tyrosine kinase family, was developed and validated. Plasma samples were pre-treated using salting-out assisted liquid-liquid extraction (SALLE) with acetonitrile, magnesium chloride a stable isotopically labeled internal standard. After dilution, extract directly injected into reversed-phase chromatographic system. The eluate transferred electrospray interface positive ionization compounds detected in selected reaction monitoring mode triple quadrupole spectrometer. completely validated plasma 0.5-500ng/ml calibration range r(2)=0.995±0.002 (n=6) linear regression inverse square concentration as weighting factor calibration. Within-run precisions (n=18) 2.7-11.7% between-run (3 runs; n=18) 3.0-14.5%. Accuracies between 96-109% whole range. drug sufficiently under all relevant analytical conditions. Finally, successfully applied to determine levels study pharmacokinetics after oral administration afatinib female FVB mice.