A new stability indicating RP-HPLC method for the simultaneous estimation of Diloxanide and Ornidazole in bulk and Pharmaceutical Dosage forms

Abstract

A simple, accurate and precise stability indicating RP-HPLC method for the simultaneous estimation of Diloxanide and Ornidazole in pure and pharmaceutical dosage form has been reported. Chromatography was performed with Shimadzu HPLC equipment comprisingdiscovery (250 mm × 4.6mm, 5µ) column with photodiode array detector. A Rheodyne injector fitted with a 10 µl loop was also used and data was recorded and evaluated using LC-20 solutions software. The mobile phase consists of 50 % of ortho phosphoric acid buffer and 50 % acetonitrile at a flow rate of 1 ml/min. The Diloxanide and Ornidazole were eluted at approximately 2.516 minutes and 3.973 minutes, respectively. The wavelength was found to be 260 nm. A linear response was observed at concentration range of 81.25-487.5 µg/ml for Diloxanide and 62.5-375 µg/ml for Ornidazole with a regression coefficient of 0.9992 and 0.9993, respectively. Forced degradation studies were performed on pure sample of Diloxanide and Ornidazole using acid (0.1 Normal (N) Hydrochloric acid), base (0.1 N sodium hydroxide), peroxide (30% H2O2), thermal (105°C) conditions, UV (25°C) conditions and neutral conditions. The developed method was validated with respect to specificity, precision (% RSD about 0.4%), robustness, LOD and LOQ values were found to be 0.42 µg/ml and 1.28 µg/ml for Diloxanide and 0.42 µg/ml and 1.27 µg/ml for Ornidazole, respectively.