ASSAY METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTITATIVE ESTIMATION OF DILOXANIDE FUROATE AND TINIDAZOLE IN TABLETS BY RP-HPLC
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.
Method: The optimized method uses a reverse phase column, Waters Symmetry C18 (250 X 4.6 mm;5Î¼), a mobile phase of triethylammonium phosphate buffer (pH 2.3):acetonitrile in the proportion of 40:60 v/v, flow rate of 1.0ml/min and a detection wavelength of 270 nm using a UV detector.
Results: The developed method resulted in Diloxanide furoate eluting at 4.07min and Tinidazole at 2.52min. Diloxanide furoate exhibited linearity in the range 31.25-93.75Î¼g/ml, while Tinidazole exhibited linearity in the range 37.5-112.5Î¼g/ml. The precision is exemplified by relative standard deviations of 0.90% for Diloxanide furoate and 0.68% 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 68.53Âµg/ml and 97.87Âµg/ml respectively, while limit of quantitiation (LOQ) for Diloxanide furoate and Tinidazole were found to be 207.677Âµg/ml and 296.6Âµg/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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