ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF OFLOXACIN AND METRONIDAZOLE IN BULK AND DOSAGE FORM USING RP-HPLC

A new method was established for simultaneous estimation of Ofloxacin and Metronidazole by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Ofloxacin and Metronidazole by using Agilent C18 5µm (4.6*250mm) column, flow rate was 1ml/min, mobile phase ratio was Methanol: ACN (70:30%v/v), detection wave length was 238nm. The instrument used was WATERS HPLC Auto Sampler, Separation module 2695, PDA Detector 996, Empower-software version-2.The retention times were found to be 2.443 mins and 2.918 mins. The % purity of Ofloxacin and Metronidazole was found to be 100.7% and 101.4% respectively. The system suitability parameters for Ofloxacin and Metronidazole such as theoretical plates and tailing factor were found to be 1.7, 2114.5and 1.7, 2931.0 the resolution was found to be 8.0.The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study for Ofloxacin and Metronidazole was found in concentration range of 1μg-5μg and 100μg-500μg and correlation coefficient (r2) was found to be 0.999 and 0.999, % mean recovery was found to be 100% and 100.5%, %RSD for repeatability was 2.0 and 2.0, % RSD for intermediate precision was 1.5 and 1.1 respectively. The precision study was precise, robust, and repeatable. LOD value was 2.95 and 3.04, and LOQ value was 9.87 and 10 respectively. Hence the suggested RP-HPLC method can be used for routine analysis of Ofloxacin and Metronidazole in API and Pharmaceutical dosage form. Keywords: Agilent C18, Ofloxacin and Metronidazole, RP-HPLC method.