Development and validation of UPLC method for simultaneous estimation of Darunavir, Cobicistat, Emtricitabine and Tenofovir alafenamide in bulk drug and pharmaceutical dosage form

Abstract

The main objective of the present study was to develop and validate a multicomponent analysis of Darunavir, Cobicistat, Emtricitabine and Tenofovir alafenamide in bulk drug and pharmaceutical dosage form by ultra-performance liquid chromatography (UPLC). The drugs were separated using HSS C18 (100mm × 2.1mm, 1.8µ particle size) column with mobile phase consisting of potassium dihydrogen phosphate, pH adjusted to 5.4 and acetonitrile in the composition of 60:40 %v/v operated in isocratic mode at a flow rate of 0.3 ml/min. The column temperature maintained at 30˚C and detection wavelength used was 260 nm. The retention time for Darunavir was found to be 1.04min, for Cobicistat it was 1.82 min, for Emtricitabine it was 2.28 min and for Tenofovir it was found to be 1.39min. The developed method was validated in accordance to the ICH guidelines. The method obeyed Beer’s law in the concentration range of 50 µg/ml- 300µg/ml for Darunavir, 7.5 µg/ml – 45 µg/ml for Cobicistat, 25µg/ml - 150µg/ml for Emtricitabine and 0.625 µg/ml – 3.75µg/ml for Tenofovir alafenamide, with correlation coefficient of 0.999. The % relative standard deviation for all the drugs was found to be less than two indicating the method to be precise. The mean % recovery was found to be within the limits for all the drugs indicating that the method to be accurate. The method was also found to be specific, robust and stable. From the forced degradation studies, it was concluded that the drugs were found to be stable when exposed to different stress conditions as the net degradation was found within the acceptance criteria. The developed method for the multicomponent analysis of Darunavir, Cobicistat, Emtricitabine and Tenofovir alafenamide using UPLC can be used for quality control and routine analysis in bulk drug and pharmaceutical dosage form.