Development, Validation and Stress Degradation Studies of Emtricitabine and Tenofovir Disoproxil Fumerate by High Performance Liquid Chromatography

A simple, selective, rapid, precise and accurate reverse phase high pressure liquid chromatographic method has been developed for simultaneous estimation of Emtricitabine and Tenofovir disoproxil fumerate in pharmaceutical Tablet dosage form. The mobile phase consisted of 65:35 % (v/v) of Methanol and 0.1M of potassium di hydrogen ortho phosphate and pH adjusted to 3.2 with ortho phosphoric acid.The method developed is operated on isocratic mode.The flow rate is 1.0 ml/min. Chromatographic determination of Emtricitabine and Tenofovir disoproxil fumerate was performed on Phenomenex C18 column (150 X 4.6 mm Id, ODS-2, 5μm). The wavelength of detection is 260 nm. The injection volume is 20μL. The retention time of Emtricitabine is 1.92 ± 0.01 minutes while the retention time of Tenofovir disoproxil fumerate is 3.17 ± 0.01minures.The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantitation, solution stability, ruggedness, and robustness. The influence of Acid, Alkaline, Oxidative Stress, Photolytic stress conditions on Emtricitabine and Tenofovir disoproxil fumerate was studied. Results indicated that Emtricitabine and Tenofovir disoproxil fumerate is stable under the experimental conditions. The proposed method has been successfully used for the estimation in tablet dosage forms.