Development and Validation of a Stability-indicating Method for Neostigmine Bromide: Separation, Identification and Characterization of Degradation Product using LC-MS and ESI-Q-TOF-MS/MS

Abstract: Objectives: Forced degradation study of the drug product and substance is very much important in development discovery to establish intrinsic stability understand its behaviors towards different stress conditions. In present work, testing neostigmine bromide (NBr) was carried out as per ICH guidelines identify characterize (DP) formed. Materials Methods: According guidelines, subjected various Waters BEH (Ethylene Bridge Hybrid) C-18 column (1.7 m, 100 mm 2.1 mm) having composition mobile phase Eluent A: 0.01M KH2PO4 water pH-2.5 with orthophosphoric acid B: Acetonitrile utilizing 220 nm wavelengths provided best separation DP component. Throughout analysis, injection volume (5 μL) flow rate (0.3 mLmin -1) were kept constant. Results: The limits detection (LOD) quantitation (LOQ) set at 25 ngmL-1 50 ngmL-1, respectively. method showed excellent linearity over a concentration range 25-250 regression coefficient (R2) value 0.9999. results that significant observed base oxidative conditions whereas found neutral, acidic, photolytic thermal Conclusion: identified characterized using sophisticated HRMS/MS/TOF approach for accurate mass measurement ESI positive mode ionization. study, establishment pathway fragmentation DP-I explained which never reported any literature. Keywords: Neostigmine bromide, RP-UPLC, Stability-indicating, Stress Validation.