Green Analytical HPLC Method Development for Impurity Profiling of Ivosidenib with LCMS/MS Characterization and In Silico Toxicity Prediction of its Degradation Products

Abstract

The study aimed to optimize and validate a robust green HPLC method for quantifying and separating process-related impurities of ivosidenib. Further, the study intended to characterize the degradation products (DPs) of ivosidenib formed under various stress conditions. Among the columns tested, the Zorbax Eclipse Plus C18 column (250 mm) column was proved to be the best for the resolution of ivosidenib and its impurities using ethanol and 0.1% aqueous formic acid in 45:55 (v/v) at 0.8 mL/min flow rate and 245 nm detector wavelength. This method produces a clear resolution of ivosidenib and its impurities with no interference from blank samples providing the specificity and robustness of the method. The technique demonstrated linearity (r² ≥ 0.9994), precision (intraday and interday % RSD < 2), accuracy (recovery: 98–102%), ruggedness, robustness, and stability under varied conditions. Forced degradation studies revealed distinct DPs under acidic, UV, and oxidative stress conditions, and these DPs were characterized using LC–MS techniques. The toxicity prediction indicated neurotoxicity and respiratory toxicity for all DPs with specific compounds exhibiting hepatotoxicity, nephrotoxicity, or immunotoxicity. The environmental impact of the proposed method was assessed through AGREE and GAPI tools and the technique exhibits high compliance with green analytical chemistry principles. This result highlights method reliability, efficiency, and eco-friendliness to pharmaceutical safety and environmental sustainability.