A rapid, simple, sensitive and selective analytical method was developed by using reverse phase Ultra performance liquid chromatographic technique for the simultaneous estimation of darunavir, dolutegravir and ritonavir in combined tablet dosage form. The developed method is superior in technology to conventional high performance liquid chromatography with respect to speed, resolution, solvent consumption, time, and cost of analysis. Elution time for the separation was 3 min and ultra violet detection was carried out at 225 nm. Efficient separation was achieved on BEH C18 (50 x 2.1mm, 1.7 ) UPLC column using phosphate buffer and acetonitrile as organic solvent in a linear isocratic program. Resolution between darunavir, dolutegravir and ritonavir was found to be more than 3.5. The active pharmaceutical ingredient was extracted from tablet dosage from using a mixture of water and acetonitrile used as diluent. The calibration graphs were linear for darunavir, dolutegravir and ritonavir in the range of 100-600 g/ml, 12.5-75 g/ml and 6.25-37.5 g/ml respectively. The mean percentage recoveries for darunavir, dolutegravir and ritonavir were found to be in the range of 99.13 and 99.59 and 99.72 % respectively. Developed UPLC method was validated as per International Conference on Harmonization specifications for method validation. This method can be successfully employed for simultaneous estimation of darunavir, dolutegravir and ritonavir in bulk drugs and formulations.
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