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Increasing the Throughput and Productivity of Caco-2 Cell Permeability Assays Using Liquid Chromatography-Mass Spectrometry: Application to Resveratrol Absorption and Metabolism

[ Vol. 6 , Issue. 8 ]


Yongmei Li, Young Geun Shin, Chongwoo Yu, Jerome W. Kosmeder, Wendy H. Hirschelman, John M. Pezzuto and Richard B. van Breemen   Pages 757 - 767 ( 11 )


The Caco-2 cell monolayer permeability assay has become a standard model of human intestinal absorption and transport. This paper reviews recent progress in increasing the throughput of Caco-2 cell monolayer assays and in expanding the scope of this assay to include modeling intestinal drug metabolism. The state-of-the-art in Caco-2 cell monolayer permeability assays combines multi-well plates fitted with semipermeable inserts on which Caco-2 cells have been cultured with liquid chromatography-mass spectrometry (LC-MS) or LC-tandem mass spectrometry (LC-MS-MS) for the quantitative analysis of test compounds and the identification of their intestinal metabolites. After reviewing the progress in increasing the throughput of Caco-2 cell monolayer assays for both modeling human intestinal permeability or transport and the metabolism of xenobiotic compounds, we demonstrate the application of LC-MS and LC-MS-MS to the measurement of resveratrol permeability and metabolism in the Caco-2 model. trans-Resveratrol (trans-3,5,4- trihydroxystilbene) is a polyphenolic compound occurring in grapes, peanuts and other food sources, that is under investigation as a cancer chemoprevention agent. The apparent permeability coefficient for apical (AP) to basolateral (BL) movement of resveratrol was 2.0 × 10 -5cm/sec. Resveratrol was not a substrate for Pglycoprotein or the multi-drug resistance associated proteins (MRP). No phase I metabolites were observed, but the phase II conjugates resveratrol-3-glucuronide and resveratrol-3-sulfate was identified based on LC-MS and LC-MS-MS analysis and comparison with synthetic standards. Although these data indicate that resveratrol diffuses rapidly across the intestinal epithelium, extensive phase II metabolism during absorption might reduce resveratrol bioavailability.


resveratrol, caco-2 cells, mass spectrometry, permeability, metabolism, resveratrol-3-glucuronide, resveratrol-3- sulfate


Department of Medicinal Chemistry and Pharmacognosy, University of Illinois College of Pharmacy,833 S. Wood Street, Chicago, IL 60612 USA.

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