Development and Validation of RP-HPLC Method for Simultaneous Determination of Ofloxacin and Ornidazole In Infusion

A simple reverse phase liquid chromatographic method has been developed and subsequently validated for simultaneous determination of Ofloxacin and Ornidazole in infusion dosage form. The separation was carried out using a mobile phase containing methanol and buffer (equal proportion of 0.01M orthophosphoric acid and 0.01M sodium phosphate monobasic dihydrate) with pH 4.00 adjusted by 20% of triethylamine in the ratio of 60:40 v/v. The column used was HiQ Sil C18 (150 mm x 4.6 mm i.d, 5 µ) with flow rate of 1 mL / min using UV detection at 300 nm. The described method was linear over a concentration range of 1.25-10 µg/mL (r2>0.9991) for Ofloxacin and 3.12-25 µg/mL (r2>0.9992) for Ornidazole. Separation was achieved within 5 min. The mean % recovery was found to be 99.94% for Ofloxacin and 100.27 % for Ornidazole. The limit of detection (LOD) for Ofloxacin and Ornidazole were found to be 0.146 and 0.25 µg/mL respectively. Whereas the limit of quantification (LOQ) for Ofloxacin and Ornidazole was 0.44 and 0.77 µg/mL respectively. The results of the study showed that the proposed RP-HPLC method is simple, rapid, precise, accurate and cost effective which is useful for the routine determination of Ofloxacin and Ornidazole in bulk drug and in its infusion.