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

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

Volume: 6 Issue: 8

Author(s): Yongmei Li, Young Geun Shin, Chongwoo Yu, Jerome W. Kosmeder, Wendy H. Hirschelman, John M. Pezzuto and Richard B. van Breemen

Affiliation:

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

Abstract: 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.

Cite this article as:

Li Yongmei, Shin Geun Young, Yu Chongwoo, Kosmeder W. Jerome, Hirschelman H. Wendy, Pezzuto M. John and van Breemen B. Richard, Increasing the Throughput and Productivity of Caco-2 Cell Permeability Assays Using Liquid Chromatography-Mass Spectrometry: Application to Resveratrol Absorption and Metabolism, Combinatorial Chemistry & High Throughput Screening 2003; 6 (8) . https://dx.doi.org/10.2174/138620703771826865