A reverse phase liquid chromatography (LC) method was developed and validated for simultaneous estimation of closantel and ivermectin in injection dosage form. The isocratic LC analysis was performed on phenomenex gemini ODS C18 column (200 mm x 4.6 mm, 5 μ) using mobile phase composed of acetonitrile : water pH 3 adjusted by orthophosphoric acid (80:20, v/v) at a flow rate of 1.0 ml/min. Quantitation was performed using UV detector at 240 nm. The retention times were found to be 3.56±0.004 min for closantel and 5.46±0.007 min for ivermectin. The analytical method was validated according to ICH guidelines. The linearity was observed in the range of 25-125 and 1-5 μg/ml with correlation coefficient, r2 = 0.999 and 0.999 for closantel and ivermectin respectively. The accuracy (%recovery) was found to be 99.27 - 100.13 % for closantel and 99.00–100.18 % for ivermectin. The relative standard deviation values for repeatability and intermediate precision studies were less than 2%. The method was successfully applied for market sample analysis and mean percentage assay values were 99.97±0.87 and 99.09±0.22 for closantel and ivermectin respectively. The present method is precise and accurate and can be used for the routine estimation of closantel and ivermectin in injection dosage form.
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