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J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Oct 15;878(28):2789-95. doi: 10.1016/j.jchromb.2010.08.023. Epub 2010 Aug 21.

Application of one-step liquid chromatography-electrospray tandem MS/MS and collision-induced dissociation to quantification of ezetimibe and identification of its glucuronated metabolite in human serum: A pharmacokinetic study.

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  • 1Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.


A new one-step liquid chromatography-electrospray tandem MS/MS method is described to quantify ezetimibe (EZM) a novel lipid lowering drug in human serum. Also using collision-induced dissociation (CID) of the analyte, identification and chromatographic separation of its major metabolite, ezetimibe glucuronide (EZM-G) is achieved in this study. A thawed serum aliquot of 100μL was deproteinated by addition of 500μL methanol containing omeprazole as internal standard (I.S.). Separation of the drug, its metabolite and the I.S. were achieved using acetonitrile-water (70:30, v/v) as mobile phase at flow rate of 0.5mL/min on a MZ PerfectSil target C18 column. Multiple reaction monitoring (MRM) mode of precursor-product ion transition (408.7→272.0 for EZM and 345→194.5 for the I.S.) was applied for detection and quantification of the drug while, EZM-G was chromatographically separated and identified using CID. The analytical method was linear over the concentration range of 1-32ng/mL of EZM in human serum with a limit of quantification of 1ng/mL. The coefficient variation values of both inter- and intra-day analysis were less than 8% whereas the percentage error was less than 3.7. The validated method was applied in a randomized cross-over bioequivalence study of two different EZM preparations in 24 healthy volunteers.

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