Development and Validation of a Novel UPLC-MS/ MS Method for Quantification of Tovorafenib and Characterization of its Forced Degradation Products

Abstract

Aim and Objectives: The current study was primarily aimed at the development of a validated stability indicating UPLC method for tovorafenib and its related impurities, and UPLC-MS/MS analysis of degradation products. Materials and Methods: The optimized chromatographic conditions included a phenyl column (2.1×100 mm, 1.7 µm) with a flow rate of 0.5 mL/min, an injection volume of 5 µL, and a mobile phase of a 30:70 ratio of acetonitrile and 0.1% formic acid buffer. The detection wavelength was established at 260 nm and the run time of the detection was 6.0 min. The developed approach was validated according to the ICH recommendations. Tests for degradation were carried out and MS/MS analysis was used to identify degradation impurities. Results and Discussion: The impurities and tovorafenib both fulfilled the required accuracy thresholds. The correlation co-efficient was observed to be more than 0.999. In terms of robustness and ruggedness, the % RSD was less than 2. The degradation products were discovered during acidic, alkaline, peroxide, and reduction analyses. The UPLC-MS/MS analysis characterized the fragmented ions formed from degradation product which is parent ion. Conclusion: The created approach complied with the ICH guidelines reliability requirements. Additionally, it used UPLC-MS/MS to successfully characterize degrading impurities.