Development and Validation of RP-HPLC for the Rabeprazole sodium in Pharmaceutical Formulations and Human Plasma

A simple, selective, accurate high Performance Liquid Chromatographic (HPLC) method was developed and validated for the analysis of Rebeprazole sodium. Chromatographic separation achieved isocratically on a C₁₈ column [Use Inertsil C₁₈, 5 μ, 150 mm×4.6 mm] utilizing a mobile phase of acetonitrile/phosphate buffer (70:30, v/v, pH 7.0) at a flow rate of 0.8 ml/min with UV detection at 228 nm. The retention time of Rabeprazole sodium was 2.44 min. The method is accurate (99.15-101.85%), precise (intra-day variation 0.13-1.56% and inter-day variation 0.30-1.60%) and linear within range 0.1-30 μg/ml (R²=0.999) concentration and was successfully used in monitoring left over drug. The detection limit of Rabeprazole sodium at a signal-to-noise ratio of 3 was 1.80 ng/ml in human plasma while quantification limit in human serum was 5.60 ng/ml. The proposed method is applicable to routine analysis of Rabeprazole sodium in pharmaceutical formulations as well as in human plasma samples.