RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTITATIVE ESTIMATION OF DILOXANIDE FUROATE AND TINIDAZOLE IN TABLETS
Keywords:RP-HPLC, Diloxanide furoate, Tinidazole, Method development, Validation
Objective: To develop an accurate, precise and linear Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for simultaneous quantitative estimation of Diloxanide furoate and Tinidazole in tablets and validate as per ICH guidelines.
Methods: The optimized method uses a reverse phase C18 column, ZODIAC (250 X 4.6 mm; 5Î¼), mobile phase consisting of mixed phosphate buffer (pH 6.5; KH2PO4+K2HPO4): acetonitrile in the proportion of 30:70 v/v. The mobile phase was set at a flow rate of 1.0 ml/min and the volume injected was 20Î¼l for every injection. The detection wavelength was set at 270 nm.
Results: The developed method resulted in Diloxanide furoate eluting at 4.70 min and Tinidazole at 3.45 min. Diloxanide furoate exhibited linearity in the range 30-70Î¼g/ml, while Tinidazole exhibited linearity in the range 36-84Î¼g/ml. The precision is exemplified by relative standard deviations of 0.266% for Diloxanide furoate and 0.35% for Tinidazole. Percentage Mean recoveries were found to be in the range of 98â€102, during accuracy studies. The limit of detection (LOD) for Diloxanide furoate and Tinidazole were found to be 0.32Âµg/ml and 0.40Âµg/ml respectively, while limit of quantitiation (LOQ) for Diloxanide furoate and Tinidazole were found to be 0.96Âµg/ml and 1.21Âµg/ml respectively.
Conclusion: A simple, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of Diloxanide furoate and Tinidazole in tablets and validated as per ICH guidelines. Hence it can be used for the routine analysis of Diloxanide furoate and Tinidazole in tablets in various pharmaceutical industries.
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