A novel, precise, accurate and rapid isocratic reversed-phase high performance liquid chromatographic/ultraviolet (RP-HPLC/UV) method was developed, optimized and validated for simultaneous determination of Ofloxacin and Flavoxate hydrochloride. Effect of different experimental parameters and various particulate columns on the analysis of these analytes was evaluated. The method showed adequate separation for Ofloxacin and Flavoxate hydrochloride and best resolution was achieved with Kromasil analytical C18, (250 mm × 4.6 mm × 5 μm) using acetonitrile-methanol-water (30:20:50, v/v; pH adjusted to 3.0 with TEA and o-phosphoric acid) as a mobile phase at a flow rate of 1ml/min and wavelength of 322 nm. The calibration curves were linear over the concentration ranges of 10-60 μg/ml for Ofloxacin and 10-60 μg/ml for Flavoxate hydrochloride. The limit of detection (LOD) and limit of quantification (LOQ) for Ofloxacin were 0.239 and 0.724 μg/ml while for Flavoxate hydrochloride were 0.496 and 1.506 μg/ml, respectively. All the analytes were separated in less than 7.0 min. The proposed method could be applied for routine laboratory analysis of Ofloxacin and Flavoxate hydrochloride in pharmaceutical dosage form.
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