Validated liquid chromatographic method for quantitative determination of Rufinamide active pharmaceutical ingredient form and its impurities
Aim: A new reversed-phase liquid chromatography method has been developed for the determination of Rufinamide in active pharmaceutical ingredient form. Methods: The chromatographic column used is Inertsil ODS 3V, GL Sciences Inc. C18, dimensions (250 mm × 4.6 mm, 5 mm), on Waters 2487 HPLC system. The mobile phase is used in a gradient programme where mobile phase A is 0.1% o-phosphoric acidin water and mobile phase B is a premixed solution of methanol, acetonitrile and tetrahydrofuran in the ratio of 900:70:30 v/v/v. The flow rate applied for the method is 1.0 ml/min and detection wavelength employed is 220 nm. The retention time of Rufinamide API Main peak was found to be 21.3 min. The linearity has been tested for impurities and API over concentration range of Limit of quantitation (LOQ), 0.75 mg/ml to 2.25 mg/ml for Impurity A and LOQ, 0.25 mg/ml to 0.75 mg/ml for Impurity B and LOQ, 0.125 mg/ml to 0.375 mg/ml for both Rufinamide and Impurity C, and the Resulting correlation coefficient were found to be greater than 0.99. The percentage recoveries were found to lie within 80%–120% for LOQ level and 90%–110% for other levels. The method has been validated in accordance with International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Conclusion: The proposed validated method has been applied for the quantitative analysis of Rufinamide in API form and its impurities, which will help to improve quality Control. Key words: Analytical method validation,API,Gradient programme,Rufinamide.