Abstract

Development and validation of RP-HPLC method for simultaneous estimation of gemifloxacin mesylate and ambroxol hydrochloride in pharmaceutical formulation

Author(s): Sagar B.Wankhede, Anand M.Mahajan, Sohan S.Chitlange

A simple, precise, sensitive, and validated reverse phase high performance liquid chromatographic method has been developed and validated for simultaneous estimationGemifloxacinmesylate (GFM) andAmbroxol hydrochloride (AMB) in tablets. Chromatographic seperation was performed on agilent ODS C18 (250 × 4.6 mm i.d., 5µm) column with a mobile phase comprising mixture of 25 mM potassium dihydrogen orthophosphate buffer (pH 3.5, adjusted with orthophosphoric acid) : acetonitrile (75:25 v/v) at a flowrate 1 ml/min, withUV-detection at 246 nm. Separationwas completed in less than 10 min and retention time for GFM and AMB by proposed method was found to be 7.38 and 8.73 min, respectively. The validated calibration range for GFMandAMB was 64-320 µg/ml and 24-120 µg/ml, respectively. The LODswere 2.920 and 0.767 µg/mland the LOQswere 8.85 and 2.32 µg/mlforGFMandAMB, respectively. The suitability of thisHPLC method for quantitative determination of the compounds was proved by validation in accordancewith the ICH guidelines. Statistical analysis proved that the method was accurate, precise, and reproducible. To establish stability indicating nature of the LC method, forced degradation of drug substances was performed under different stress conditions viz. acid and base hydrolysis, dry heat degradation and oxidation. Since the method is able to selectively quantitate these drugs in presence of their degradation products it can be used as a stability indicating method.