Development and Validation of RP-HPLC Method for Simultaneous Estimation of Sparfloxacin and Dexamethasone in Bulk and Pharmaceutical Dosage Form

In the present work a simple, rapid reverse phase high performance liquid chromatographic method (RP-HPLC) has been developed and validated for simultaneous estimation of Sparfloxacin and Dexamethasone in pharmaceutical dosage form. Chromatographic separation was carried out on Shimadzu RP-HPLC instrument (LC-20AT) using C-18 (250 mm × 4.6 mm, 5.0μ) as stationary phase and mobile phase containing 0.02m Phosphate Buffer pH (3.5): Acetonitrile (60:40 v/v) at flow rate of 1 ml/min using UV detection at 247 nm. The retention time for Sparfloxacin and Dexamethasone was found to be 5.773 and 7.420 min respectively. The method was validated as per ICH guideline and successfully used for the quantitative analysis of commercially available Eye drops. The calibration curve was linear over the concentration range of 3–45μg/ml for Sparfloxacin and 1–15μg/ml for Dexamethasone. Limit of detection and quantitation was found to be 0.5112 μg/ml and 1.549μg/ml for Sparfloxacin and 0.4912μg/ml and 1.488μg/ml for Dexamethasone respectively. The % RSD below 2.0 shows the high precision of proposed method.