DETERMINATION AND CHARACTERIZATION OF PROCESS IMPURITIES IN PAZOPANIB HYDROCHLORIDE DRUG SUBSTANCE
Keywords:Nil, Pazopanib hydrochloride, Method development, Validation
Objective: To develop a rapid, sensitive, accurate, precise and linear Reverse-Phase High-Performance Liquid Chromatographic (RP-HPLC) method and validate as per ICH guidelines for the quantitative estimation of Pazopanib Hydrochloride (PBH) drug substance along with two process impurities.
Methods: The developed method uses a reverse phase Zorbax RP18 column (50 mmÃ—4.6 mm; 5.0 Âµm), a mobile phase of 0.015M Potassium dihydrogen orthophosphate buffer (pH 3.0 with orthophosphoric acid) and methanol in the proportion of 16:84 v/v. The mobile phase was set at a flow rate of 0.8 ml/min and the volume injected was 10Î¼l for every injection. The detection wavelength was set at 215 nm.
Results: Newly developed method resulted in eluting the two process impurities of PBH at 2.998 and 23.548 min respectively. The detection limits (LOD) were about 0.0061 and 0.0062 mg/ml and quantitation limit (LOQ) were about 0.024 and 0.021 mg/ml. The relative standard deviation was found to be 0.78 % and 1.38 % respectively for the two process impurities of PBH. The % recovery of the PBH impurities ranged from 97.3 to 100 % and 94.0 % to 99.0 % respectively.
Conclusion: Two new process impurities of PBH drug substance were determined and characterized by newly developed RP-HPLC method, and identified using LCâ€“MS technique. Proposed structures of these impurities were confirmed by structural elucidation using NMR techniques. The HPLC method was validated as per ICH guidelines. The newly developed reversed-phase liquid chromatographic method was found to be accurate, simple, sensitive and selective. It was also found to exhibit excellent resolution for the two process impurities and the PBH drug substance indicating high sensitivity and selectivity of the validated method.Keywords: RPâ€HPLC, Pazopanib hydrochloride, Method development, Validation
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