Design of Experiment Based Validated Stability Indicating Method for Determination of Besifloxacin Hydrochloride in Bulk Drug and Formulation

Abstract

A simple, precise, stability indicating RP-HPLC method was developed and validated for determination of Besifloxacin HCl (Besi) in bulk sample and application of that method for estimation of drug i.A Box-Behnken statistical design with three factors and five levels was employed to optimize the chromatographic conditions. The separation was achieved using solvent system combination using methanol:Acetonitrile: phosphate buffer pH 3(25:25:50). The developed method was validated for linearity, precision, accuracy, limit of detection and limit of quantitation, and robustness in accordance with the ICH guidelines. The method was found linear with regression coefficient of over a wide range of 2.5-15 μg/mL. All the validation parameters were found to be within the acceptance range. The developed method was successfully applied to estimate the amount of Besifloxacin HCl in pharmaceutical formulations.