A Novel Stability Indicating Method Development and Validation of Olutasidenib in HPLC and Characterisation of New Degradation Products by LC-MS/MS

A simple, stable, economic, and less time-consuming HPLC method was developed for the separation and quantification of Olutasidenib and its related impurities. Chromatographic separation was achieved using a Waters X-Terra RP-۱۸ column (۱۵۰ mm × ۴.۶ mm, ۳.۵ µm). The mobile phase consisted of acetonitrile and ammonium acetate buffer (pH ۲.۵ adjusted with ۰.۱% formic acid) operated in gradient mode with a time/buffer composition of ۰/۵۰, ۵/۳۰, ۸/۵۰, and ۱۲/۵۰ (T/B). The flow rate was maintained at ۱.۰ mL/min with detection at ۲۴۹ nm and an injection volume of ۱۰ µL. The total runtime of the method was ۱۲ minutes, enabling efficient separation of Olutasidenib and its related impurities. The method exhibited good linearity in the concentration range of ۳۷.۵۰–۲۲۵.۰۰ µg/mL for Olutasidenib and ۱.۲۵–۷.۵۰ µg/mL for its impurities, with validation parameters complying with analytical guidelines. The sensitivity of the method was evaluated using the signal-to-noise approach, where the limit of detection (LOD) and limit of quantification (LOQ) for impurities were determined as ۰.۱۵ µg/mL and ۰.۵ µg/mL, respectively. Forced degradation studies demonstrated that Olutasidenib remained stable under acidic, neutral hydrolytic, and photolytic conditions, while significant degradation was observed under alkaline, oxidative, thermal, and reducing conditions, resulting in the formation of degradation products. The developed method proved to be robust and stability-indicating, making it suitable for routine quality control and stability analysis of Olutasidenib and its related impurities in pharmaceutical formulations.

کلیدواژه ها:

Olutasidenib ، Related impurities ، Forced Degradation studies ، Fragmentation pathway