A simple, economic, selective, precise and stability indicating reverse HPLC method has been developed and validated for ampicillin and sulbactam in injection dosage form. The isocratic LC analysis was performed on Phenomenex Gemini ODS C18 column (200 mm x 4.6 mm, 5μ) using mobile phase composed of acetonitrile: buffer pH 3.5 adjusted by phosphoric acid (60:40, v/v) at a flow rate of 1.0 ml/min. Quantitation was performed using UV detector at 250 nm. The retention times were found to be 3.263 min for sulbactam and 7.147 min for ampicillin. The analytical method was validated according to ICH guidelines. The linearity was observed in the range of 10-30 and 5-15 μg/ml with correlation coefficient, r=0.995 and 0.999 for ampicillin and sulbactam respectively. The accuracy (% recovery) was found to be 98.95 - 100.30 % for ampicillin and 99.17-100.50 % for sulbactam. 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 100.50 and 98.65 for ampicillin and sulbactam respectively. Ampicillin and sulbactam active substance was subjected to forced degradation to demonstrate the stability indicating power of the HPLC method. Acidic, basic, photolytic, thermal and oxidative degradation were used to assess the stability of the method. The drug was found to degrade in all studied conditions but the extent of degradation was different. The proposed method is rapid, cost-effective and can be used as a quality-control tool for routine quantitative analysis of ampicillin and sulbactam. The present method is precise and accurate and can be used for the routine estimation of ampicillin and sulbactam in injection dosage form.
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