Stability indicating method development and validation for the simultaneous estimation of Fluorometholone and Neomycin in bulk and Pharmaceutical dosage forms by RP-HPLC

Abstract

A Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method has been developed and validated for simultaneous estimation of Fluorometholone and Neomycin in bulk and pharmaceutical formulations. Separation was carried out using column Enable C18 (250 mm x 4.6mm x 5μm particle size) in isocratic mode using mobile phase composition Acetonitrile and 0.1% Orthophosphoric acid (pH 3.5) in the ratio 70:30 v/v and UV detection at 239 nm. The compounds were eluted at a flow rate of 0.8 ml/ min. The average retention times for Fluorometholone and Neomycin were 4.336 and 3.770 min, respectively. The method was validated according to the ICH guidelines. The % RSD of all validation parameters found to be less than 2% indicating high degree of accuracy and precision of the proposed HPLC method. The method was linear over the concentration of 10-50 μg/ml and 20- 100 μg/ml for Fluorometholone and Neomycin respectively. The LOD and LOQ of Fluorometholone were found to be 0.625 µg/ml and 1.8950 µg/ml and of Neomycin were found to be 1.0142 µg/ml and 3.0735 µg/ml.