DEVELOPMENT AND VALIDATION OF LC/MS-MS METHOD FOR ESTIMATION OF AXITINIB FORMULATIONS BY BOX BEHNKEN DESIGN

Abstract

Several analytical chromatographic methods have published about Axitinib drug candidate, but they are cumbersome, not easy to replicate and time tedious. In this regard, there is a need to develop QbD approach of a sensitive and rapid LC-MS/MS for the estimation of Axitinib in its formulation and validation. The separation of Axitinib  was achieved using the mobile phase 10mM ammonium formate and  acetonitrile in the ratio of 30:70 v/v at the flow rate of 0.87 mL/min using  Zorbax C18 (50 mm x 4.6 mm i.d., 5µm)  column. The critical method parameters were identified and was optimized using box behnken design. The obtained model was found to be statistically significant with a probability (p) value of less than 0.05 and composite desirability of 0.781. The method performance was evaluated as per ICH guidelines with linearity ranging from 1 ng/mL to 65 ng/mL with a correlation coefficient of 0.857. The LOD and LOQ limits were found to be  300 ng/mL and 1  ng/mL, respectively. The mean recovery was in the range of  96.66 to 100.1 %. During method transfer, the method was validated and verified for targeted method performances, robustness, and system suitability.