Abstract
Background: To develop a simple and accurate method for the determination of Darunavir in human plasma by liquid-liquid extraction method with Verapamil as an internal standard using UPLC-MS/MS. Methods: The separation was achieved on Agilent, Zorbax XDB C18 (2.1 x 50 mm ID, 5 μm) column. The samples were chromatographed using the mobile phase consisting of acetonitrile and 2mM ammonium formate with 0.1% formic acid in water (70:30 v/v) at a flow rate of 0.12 ml/min in a mass spectrometer ESI chamber. The Darunavir and Verapamil were eluted at 1.35 and 1.13 min respectively with a total run time of 3 min. Results: A linear response was established at 10- 2,000 ng/ml with a correlation coefficient of 0.9919. The % accuracy was found to be 98% and 102.40%. A rapid method was developed for the determination of Darunavir in human plasma. The method was strictly validated according to the ICH guidelines.
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