METHOD DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR ESTIMATION OF GENOTOXIC IMPURITIES IN TRAZODONE HCL DRUG SUBSTANCEUSING UFLC-MS/MS (ESI)

Due of their ability to damage DNA, genotoxic impurities (GTIs) in active pharmaceutical substances need strict analytical control. Trace process-related GTIs may be present in trazodone hydrochloride, an antidepressant made using multi-step synthetic processes. Nevertheless, there is currently no approved analytical technique for the simultaneous determination of all significant GTIs related to its production. In order to quantify four process-related GTIs in trazodone hydrochloride at sub-ppb levels, a quick and selective UFLC-MS/MS approach was developed and validated. Acetonitrile-5 mM ammonium acetate buffer (pH 5.0, 70:30 v/v) was used for chromatographic separation on an Avantor ACE C18-PFP column (3 µm, 4.6 × 150 mm) under isocratic conditions. Multiple reaction monitoring was used for detection in positive ESI mode. The approach was validated over 0.060-1.200 ppb. For every impurity, linearity was attained with correlation coefficients (R²) ≥ 0.99. Across three levels, accuracy studies produced recoveries ranging from 79 to 102%. Robustness was verified with intentional changes in flow rate and column temperature, and precision satisfied acceptance standards with %RSD < 15%. When kept in a refrigerator, stock solutions stayed stable for at least three days. All GTIs were found to be below the set regulatory limits when the validated procedure was applied to commercial batches. For routine quality-control monitoring of genotoxic contaminants in trazodone hydrochloride, the established UFLC-MS/MS technique is sensitive, dependable, and appropriate.