Application of RP-HPLC Method for Simultaneous Estimation of Gatifloxacin and Flurbiprofen Sodium In Ophthalmic Formulation

A simple, accurate, precise and sensitive RP- HPLC method has been developed for the determination of Gatifloxacin and Flurbiprofen Sodium in their pharmaceutical formulation. Chromatographic separation  was carried out on InertsilODS – 3 column (250 mm ×4.6 mm, 5µm ) as stationary phase by using mobile phase consisting of 0.02 M Phosphate buffer (pH 3.5 adjusted with orthophosphoric acid ) : Methanol (80 : 20 v/v). The flow rate was 1.5 ml/min with UV-detection at 245 nm. The retention time was found to be 2.59 min for Gatifloxacin and 5.41min for Flurbiprofen Sodium. The method was validated for various parameters according to ICH guideline. The linear regression analysis data for the calibration plots showed good linear relationship in the concentration range of 30 – 90 µg/ml and 3 – 9 µg/ml and correlation coefficient was found to be 0.9988 and 0.9992 for Gatifloxacin and Flurbiprofen Sodium respectively. The Limit of Detection for Gatifloxacin and Flurbiprofen Sodium were 1.45 and 0.028 respectively. The Limit of Quantification for Gatifloxacin and Flurbiprofen Sodium were 4.39 and 0.28 respectively.