RP-HPLC Method Development and Validation for the Simultaneous Estimation of Cefoperazone and Sulbactam in Parenteral Preparation

Abstarct: A simple Reverse phase liquid chromatographic method has been developed and subsequently validated for simultaneous determination of Cefoperazone and Sulbactam in parenteral preparation. The separation was carried out using a mobile phase consisting of Phosphate Buffer pH 3.5 adjusted with ortho phosphoric acid and Acetonitrile (35:65). The column used was Kromasil C8, 5μ, 15 cm × 4.6 mm id with flow rate of 1 ml / min using PDA detection at 215 nm. The described method was linear over a concentration range of 50 250 μg/ml and 100 500 μg/ml for the assay of Sulbactam and Cefoperazone respectively. Ornidazole (50μg/ml) was used as internal standard. The retention times of Sulbactam, Cefoperazone and Ornidazole were found to be 2.3, 4.2 and 5.1min respectively. Results of analysis were validated statistically and by recovery studies. The limit of quantification (LOQ) for Cefoperazone and Sulbactam were found to be 20 and 10 μg/ml respectively. The results of the study showed that the proposed RP-HPLC method is simple, rapid, precise and accurate, which is useful for the routine determination of Cefoperazone and Sulbactam bulk drug and in its pharmaceutical dosage form.