VALIDATED STABILITY INDICATING UPLC METHOD FOR AFATINIB AND ITS IMPURITIES IN PHARMACEUTICAL DOSAGE FORM

A novel, simple, selective, rapid stability-indicating and LC-MS compatible short run time UPLC method was developed and validated for the determination of afatinib and its related substances in bulk and finished dosage form. A reverse phase gradient mode short run time method has been developed by using UPLC to separate all the five known impurities of Afatinib. The chromatographic separation of afatinib and its related impurities was achieved by using Acquity UPLC HSS PFP column 100*2.1 mm, 1.8 μm using 0.1% v/v formic acid in Milli-Q water and acetonitrile as a co-solvent with a run time of 12 minutes. The column temperature was maintained at 30°C. The mobile phase flow rate was 0.4 mL/min and the UV detection was carried out at a wavelength of 258 nm. The proposed method was validated in terms of specificity (forced degradation), linearity, accuracy, precision, robustness and ruggedness as per ICH quality guidelines Q2 (R1). Forced degradation was performed using hydrolysis, oxidation, thermal and photolytic conditions with good resolution between degradants and analyte. Degradation peaks neither co-elute with the main analyte nor with the known impurities, indicating that the proposed method as is stability indicating. Limit of quantification for all the five known impurities and analyte were in the range of 0.02 ppm to 0.05 ppm, indicating that the method is sensitive. Recovery of the impurities and analyte at various levels ranging from LOQ level to 200% level was in the range of 96.9 to 101.8%, indicating that the method was accurate and precise. Hence the validated method can be successfully applied for the quantitative estimation of afatinib and its related impurities in bulk and tablet dosage form.