Simultaneous quantification of ticagrelor and its metabolite deshydroxyethoxy ticagrelor in human plasma by ultra-performance liquid chromatography electrospray ionization-tandem mass spectrometry
Keywords:Ticagrelor, Deshydroxyethoxy ticagrelor, LC-MS/MS, Human plasma, Liquid-Liquid Extraction
A precise, highly sensitive, selective and robust liquid chromatography-mass tandem spectrometry method was developed and validated for simultaneous quantification of Ticagrelor and Deshydroxyethoxy ticagrelor in human plasma by liquid-liquid extraction. Ticagrelor d-7 and Deshydroxyethoxy ticagrelor d-7 was used as an Internal standard. The reconstituted samples were analyzed on Eclipse XDB-C8 5μm 4.6*150mm column by using Acetonitrile and 0.1% Formic acid as the mobile phase in binary mode at the flow rate of 1.0 ml/min with a chromatographic run time of 3.0 min. The calibration curve developed was linear (r2 ≥ 0.99) in the concentration range of 2.5 to 1000 ng/mL for Ticagrelor and 1.0 to 300 ng/mL for Deshydroxyethoxy ticagrelor. All the analytes were found to be stable in various stability studies. The mean recovery for Ticagrelor, Deshydroxyethoxy Ticagrelor, Ticagrelor d-7 and Deshydroxyethoxy Ticagrelor d-7 were found to be 99.7%, 96.6%, 105.9% and 101.1% respectively. The limit of detection of the method was 0.5 ng/mL for Ticagrelor and 0.2 ng/mL for Deshydroxyethoxy ticagrelor. The developed and validated assay method was successfully applied to a pharmacokinetic study in human volunteers.
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