Abstract

A STABILITY-INDICATING UPLC METHOD FOR QUANTIFICATION OF AFATINIB RELATED SUBSTANCES IN PHARMACEUTICAL DOSAGE FORMS

Kiran Kumar Ankalla* and Nageswara Rao Gollapalli

ABSTRACT

A specific, precise, rapid and reliable stability indicating UPLC method has been developed and validated for estimation of Afatinib related substances in pharmaceutical dosage forms. Chromatographic separation was achieved on an Acquity UPLC BEH Phenyl (2.1 x 100 mm, 1.7 μm) column using gradient composition of mixer of pH 6.4 Phosphate buffer and acetonitrile in the ratio of 95:05% v/v as Mobile Phase A and mixture of Acetonitrile and methanol in the ratio of 90:10% v/v as Mobile Phase B at a flow rate of 0.5 mL/min and analytes were monitored at 260 nm. The retention times of the Afatinib was about 4.09 and relative retention times of N-Oxide impurity and hydroxy impurity were about 0.61 and 0.66 respectively. The results of specificity studies indicate that there was no interference of diluent, excipients, impurities and degradation products at retention time of analyte and it is assured that the peak response was belongs to a single component only. The detector response was linear in the range of LOQ-150% level with respect to test concentration of Afatinib, N-Oxide impurity and Hydroxy impurity. Correlation coefficient (R2) was not less than 0.99 for afatinib and its two impurities. The percentage recovery of N-Oxide impurity and Hydroxy impurity were about 102.43% and 102.35% respectively. The developed method was validated for specificity, linearity, precision, accuracy, solution stability, ruggedness and stress degradation studies were monitored. Hence, the developed method was specific, rapid and cost-effective, and it can be used for routine analysis of Afatinib related substances in pharmaceutical dosage forms.

Keywords: Afatinib; N-Oxide impurity; Hydroxy impurity; UPLC.