QbD Based Development and Validation of RP-HPLC Method for Nintedanib Esylate: Application to Bioanalytical and Stability Study in Plasma

Abstract

The objective of this experiment was to develop and validate a simple, robust, and accurate Reverse- Phase High-Performance Liquid Chromatography method for estimation of Nintedanib Esylate in bulk, Pharmaceutical Dosage form, and Human Plasma. Full 3 Level Factorial design was employed for optimizing columns C8 and C18. The optimized chromatographic conditions are column C18, Ammonium format buffer: Acetonitrile (28.19:71.81 v/v) and pH of buffer (3). Furthermore, acetonitrile was used as a precipitating agent to extract Nintedanib in human plasma. After centrifuging at 2000 rpm, the supernatant was then injected and observed that the peak from blank plasma does not interfere with the peak of Nintedanib by using p-nitrophenol as an internal standard. Linearity was observed in the concentration range of 10 µg to 50 µg/mL (r2=0.9988). The accuracy range was 99.87 to 100.08 %. Intra-day and Inter-day precision was found to be 0.5432 & 0.5242 (% RSD). The bioanalytical method was validated as per ICH guideline M10 and results were found to be r2 (0.9988), Recovery (99.91 %), Specificity (99.94 %), Precision (Less than 0.543 % RSD), and Robustness (less than 0.500 % RSD. The proposed method was useful for the best analysis of Nintedanib Esylate in Bulk, pharmaceutical dosage forms and was successfully applied to a pharmacokinetic study.