BIO ANALYTICAL VALIDATED TECHNIQUE FOR THE SIMULTANEOUS ESTIMATION OF AMLODIPINE, ROSUVASTATIN AND VALSARTANIN HUMAN PLASMA BY RP-HPLC METHOD

A reverse phase high-performance liquid chromatographic bio analytical Technique (RP-HPLC) was developed and validated as per ICH 2019, US FDA 2018 guidelines for the quantification of amlodipine besylate, Rosuvastatin and Valsartan in human plasma using RP – C18 column. The mobile phase in composition (acetonitrile: water in the ratio 75:25% v/v, pH adjusted to 4.0 with acetic acid) was pumped and run at a flow rate of 0.8 ml/min, and for monitoring of eluents, a detector was set at wavelength 245 nm. The precipitation method was employed for the separation of the analyte from the plasma. A calibration curve was generated from the response and respective conc of standard solutions in the Concrange of LLOQ to ULOQ. The method was studied and validated per regulatory guidelines for selectivity, specificity, accuracy, precision, and stability study. Statistical data of the calibration curve of these drugs in the biological matrix was found within the prescribed limit. Results of accuracy and precision of quality control samples of these combined drugs were found (104.25 to 107.62 as % nominal conc. and 2.633 to 9.474 as % CV) for amlodipine, (101.01 to 106.45 as % nominal conc. and 4.431 to 13.786 as % CV) for rosuvastatin and (99.91 to 106.03 as % nominal conc. and 2.668 to 12.434 as % CV) for valsartan respectively. The developed bioanalytical method is simple: free from solvent-solvent extraction and solid phase extraction, precise, accurate and consumes less solvent due to less run time. The method suits for quantification of these drugs in plasma is henceforth applied for bioequivalence and bioavailability study in real clinical samples.