A novel validated RP-HPLC method for the simultaneous estimation of Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine in bulk andpharmaceutical tablet dosage forms

摘要

An accurate, precise, simple, efficient and reproducible, isocratic Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine in bulk and pharmaceutical tablet dosage forms. Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine were separated using an Inertsil ODS 3V C18 column (250mmtimes;4.6 mm, 5mm particle size), Waters Alliance e2695 HPLC system with 2998 PDA detector and the mobile phase contained a mixture of 0.01M Potassium dihydrogen phosphate (pH adjusted to 4 with orthophosphoric acid) and Acetonitrile (30:70, v/v). The flow rate was set to 1ml/min with the responses measured at 265nm. The retention time of Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine was found to be 1.976min, 2.661min and 4.316min respectively with resolution of 3 .1 a nd 6.8. Linearity was established for Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine in the range of 50-300mu;g/ml for Emtricitabine, 75-450mu;g/ml for Tenofovir Disoproxil Fumarate and 6.25-37.5mu;g/ml for Rilpivirine with correlation coefficient 0.999. The percentage recovery was found to be is 99.71 to 99.96, 99.68 to 100.05 and 99.82 to 100.08 for Emtricitabine, Tenofovir Disoproxil Fumarate and Rilpivirine respectively. Validation parameters such as specificity, linearity, precision, accuracy, robustness, limit of detection (LOD) and limit of quantitation (LOQ) were evaluated for the method according to the International Conference on Harmonization (ICH) Q2 R1 guidelines. The developed method was successfully applied for the quantification of bulk and active pharmaceutical ingredient present in tablet dosage form.