An LC-MS/MS method for quantification of AC0010, a novel mutant-selective epidermal growth factor receptor (EGFR) inhibitor, and its metabolites in human plasma and the application to a pharmacokinetic study.

AC0010 is an irreversible, mutant-selective EGFR inhibitor that effectively inhibits EGFR active and T790M resistance mutations in non-small cell lung cancer (NSCLC). A sensitive ultra-performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS) method was developed and fully validated for determining AC0010 and its metabolites in human plasma. The samples were purified by solid-phase extraction (SPE) columns and separated on a BEH C column. Electrospray ionization (ESI) in positive ion mode and multiple reaction monitoring (MRM) were used to monitor the ion transitions of AC0010 (m/z 488→257) and its metabolites M1 (m/z 474→403), M2 (m/z 504→487), M4 (m/z 434→377), M7 (m/z 490→405), MII-1 (m/z 651→434) and MII-2 (m/z 609→434). The results revealed that the method had excellent selectivity and linearity. Intra-day and inter-day precisions (in terms of relative standard deviation, RSD) were lower than 14.4% and the accuracies (in terms of relative error, RE) were within the range of ±10.3% for all the analytes. The lower limit of quantification (LLOQ), stability, matrix effect and extraction recovery were also validated and satisfied the criteria of validation. Finally, the method was successfully applied to a pharmacokinetic study of NSCLC patients after a single oral dose of 350mg of AC0010.