Degradation monitoring method development and validation of Mesalamine through characterization of degraded products using RP-HPLC and mass spectroscopy

Degradation monitoringRP-C18 HPLC method of Mesalamine was developed and validated using a mobile phase of acetonitrile, methanol, and water (30:50:20) at a 1.0 mL/min flow rate, with UV detection at 254 nm. The method showed linearity (25–75 µg/mL), precision, accuracy (99.72% recovery), robustness, and specificity. LOD and LOQ were 0.09 µg/mL and 0.7 µg/mL, respectively. Forced degradation under acidic, alkaline, and oxidative conditions confirmed specificity, with degradation products separated from the drug peak. Mass spectrometry identified these products via characteristic m/z fragments. Various degraded products of Mesalamine were well resoluted from the parent peak and characterized using Mass spectroscopy. The major degraded products characterized by MS include three degraded products under acid degradation, four degraded products under basic and two degraded products under oxidative stress conditions respectively. This developed method can be used for therapeutic drug monitoring to observe the degradation pathway and stability.