RP-HPLC Method for the Simultaneous Estimation and Validation of Amlodipine Besylate and Atenolol in Bulk and Tablet Dosage Form in Biorelevant Dissolution Medium (Fassif)

Abstract

Objective: A simple, rapid, and precise reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous analysis of Amlodipine Besylate and Atenolol in a tablet dosage form and in Biorelevant media has been developed and validated. Methods: The chromatographic separation was achieved using reverse phase C18 column; Kromasil C18 column (250 mm x 4.6 mm x 5µm). The mobile phase used was a mixture of Acetonitrile: Potassium di hydrogen phosphate solution (0.01M, pH 3.0 adjusting with Ortho phosphoric acid): Methanol (15:30:55) at isocratic mode and eluents were monitored at 254 nm using PDA detector. Results: By the method Amlodipine Besylate and Atenolol were eluted with retention times of 2.589 and 3.711 min, respectively. The method was continued and validated accordance with ICH guidelines. Validation revealed the method is rapid, specific, accurate, precise, reliable, and reproducible. Calibration curve plots were linear over the concentration ranges 25-125µg/mL for Amlodipine Besylate, 5-25µg/mL for Atenolol. Limits of detection (LOD) were 0.001, and 0.005µg/ml and limits of quantification (LOQ) were 0.004 and 0.015µg/mL for Amlodipine Besylate and Atenolol respectively. Conclusion: The statistical analysis was proves the method is suitable for the analysis of Amlodipine Besylate and Atenolol as a bulk and tablet dosage form in biorelevant dissolution media (Fasted State Simulated Intestinal Fluid-FaSSIF) without any interference from the excipients.