Development and validation of a novel stability indicating RP-UPLC method for the estimation of Leniolisib and characterization of its degradation products by UPLC-MS/MS

Abstract

Current analytical methods available for the quantification of Leniolisib were predominantly based on conventional HPLC and LC–MS/MS techniques, with each having inherent methodological limitations in terms of longer analysis times, laboratory efficiency and low-throughput industrial analysis. To bridge this research gap, a novel, rapid, precise and robust stability-indicating RP-UPLC method that seamlessly coupled with MS/MS has been developed for the analysis of Leniolisib. This method was designed using Acquity UPLC BEH Shield RP-18 Column (50 mm × 1.0 mm, 1.7 µm) and an isocratic mobile phase comprised of 0.015 M Ammonium acetate buffer (pH-3.0 adjusted with Formic acid): Acetonitrile in 80:20 vol/vol ratio with a flow rate of 0.2 mL/min at room temperature. Leniolisib’s retention time was found to be 1.217 min. The established method was validated statistically in accordance with the ICH criteria. Four degradation products (DP1-DP4) were identified under acid, base, oxidative and reductive stress conditions and characterized by UPLC-MS. To the best of our knowledge, this is the first stability-indicating RP-UPLC method coupled with MS/MS for the analysis of Leniolisib that enables both rapid quantification and characterization of degradants within a single analytical workflow, representing significant advancement over the existing conventional analytical approaches.