DEVELOPMENT AND VALIDATION OF RP-HPLC FOR ANALYTICAL ASSAY METHOD OF GENTAMICIN IN PHARMACEUTICAL OPHTHALMIC SOLUTION

A simple, precise, rapid and accurate RP- HPLC method was developed for the Estimation of gentamicin in pharmaceutical ophthalmic solution. An Phenomenex C18, 250 x 4.6mm dimension with 5µm particle size, the mobile phase, consisting of 0.1 % Trifluoroacetic acid and Methanol in the ratio of 70:30 v/v, prepare mixture of water and methanol used as a diluent for preparation of solution. The flow rate was 1.0 ml/min and 210 nm were used to monitor the effluents. The retention time was 4.2 for gentamicin. The detector response was linear in the concentration of 48-72 μg/mL for gentamicin. The respective linear regression equation being Y=239661x+180800 with corelation coefficient was 0.9997. The method was validated as per ICH guideline by determining its system suitability, specificity, linearity, accuracy, precision and Robustness. The results of the study demonstrated that the suggested RP-HPLC method is simple, rapid, precise and accurate, which is useful for the routine determination of gentamicin in pharmaceutical ophthalmic solution.