Quantification of Potential Genotoxic Impurities at Trace Level in Gefitinib Drug Substance by LCMS Method

A sensitive and novel reversed-phase liquid chromatography (UPLC) coupled with triple quad (TQS-micro) mass detector (LC-MS system) method was developed and validated [13] for the trace analysis of 4- (3-chloropropyl) Morpholine (KSM-02), 3-Chloro-4-fluoroaniline and 4-Chloroaniline genotoxic impurities in Gefitinib drug substance with the shorter run time. These impurities of Gefitinib were identified as genotoxic impurities using SAR / QSAR models. The method developed on waters Xevo TQS micro mass system attached with Acquity H class UPLC and Acquity UPLC BEH C18, 50x2.1mm, 1.7m column with electro spray ionization (ESI) in SIR (Single ion reaction) detection mode. The gradient mode of elution for the impurities was carried out with the aid of the mobile phase-A (0.1 % solution of formic acid in water) and mobile phase-B (Acetonitrile). The flow rate was 0.5mL/min, column oven temperature 30C and elution was monitored by mass spectrometer. The method was validated as per International Conference on Harmonization (ICH) guidelines. The method quantitates up to 0.01 ppm with respect to sample concentration of 4- (3-chloropropyl) Morpholine (KSM-02), 3-Chloro-4-fluoroaniline and 4-Chloroaniline