A high-performance liquid chromatography-tandem mass spectrometry method for the quantitative determination of four genotoxic impurities in gefitinib

In this work, a method for the determination of the amounts of the four genotoxic impurities in gefitinib has been developed. A simple and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLCMS/MS) approach has been developed, optimized, and validated. The genotoxic impurities were 3-chloro-4-fluoroaniline, 3,4-difluoroaniline, 3-fluoro-4-chloroaniline, and 3,4-dichloroaniline-Separation was achieved on an Inertsil ODS-3 column (100 mm x 3.0 mm, 3 mu m). The column temperature was 40 degrees C and the running time was 12 min. A triple quadrupole mass detector with positive electrospray ionization in the multiple reaction monitoring (MRM) mode was applied. The method was validated in terms of its specificity, linearity, precision, accuracy, stability, and robustness. The correlation coefficient of each impurity was higher than 0.999 in the range of 0.6-96.0 mu g/L. The limits of detection (LODs) and limits of quantity (LOQs) were in the ranges of 0.2-2.0 mu g/L and 0.6-6.0 mu g/L, respectively. The recoveries of all impurities at 6, 30, and 60 mu g/L were within 91.0% - 98.5%. All impurities were stable within 2 h. After detection, only 3-chloro-4-fluoroaniline was detected in the batch number 16052301 and R16052501-1 gefitinib samples, but its concentration was below the impurity limit (6 mg/L). This method is simple, reliable, and suitable for the determination of four genotoxic impurities in gefitinib. It can be further applied as a reference for quality control.