Stability-Indicating RP-UPLC Method Development and Validation for the Process Related Impurities of Nebivolol and Structural Characterization of Its Forced Degradation Products by LC-MS/MS

Abstract

The present study reports the development and validation of a stability-indicating reverse phase ultra performance liquid chromatographic method for the determination of nebivolol (NEB) in the presence of its process related impurities in bulk drugs and also demonstrates the structural characterization of forced degradation products (DP) of nebivolol by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Forced decomposition study of nebivolol was carried out and studied under acidic, basic, aqueous, oxidative, photolytic and thermal conditions according to the ICH guidelines. Nebivolol was found to be stable under photolytic, thermal and aqueous conditions applied in our study. However, nebivolol degradation was observed under basic and acidic conditions. Successful chromatographic separation of the drug and its process related impurities and degradation products were achieved on a waters acquity BEHC18 stationary phase (50 × 2.1 mm, 1.7µm) with a gradient elution of 10 mM ammonium acetate and acetonitrile at a flow rate of 0.3 mL min-1. Detection of analytes was carried out using a PDA detector with its wavelength set at 222 nm. The developed UPLC method was validated for all validation parameters according to the ICH guidelines. The degradation products were characterized by LC-MS/MS study and the most possible degradation and fragmentation pathways were proposed.