biostudies-otherUnknownDutch Research Agenda (NWA)Homo sapienshttps://www.ebi.ac.uk/biostudies/studies/S-VHPS18biostudies-otherfalseIntestinal absorption of pesticides across Caco-2 cell monolayersA monolayer of Caco-2 cells was cultured in transwells, maintaining a stable TEER for at least one week. On the apical side, cells were exposed to pesticides in HBSS (without additives) for a maximum of 5 hours. Samples were collected over time from both the basolateral and apical compartments: either 50 µL or 100 µL immediately after exposure (time point 0.01), and at 30 minutes, 1 hour, 2 hours, and 5 hours. Each sample was diluted with an equal volume (50 µL or 100 µL) of LC-MS/MS grade methanol (cyclohexane for endosulfan). Pesticide concentrations were quantified by LC-MS/MS and used to determine apical-to-basolateral transport (absorption) across the intestinal epithelial barrier. In parallel, the cytotoxicity of the pesticides towards Caco-2 cells was assessed with an alamarBlue (resazurin) mitochondrial-activity assay, to confirm that the tested concentrations did not compromise monolayer viability. The Transwell permeability procedure was adapted from Hubatsch et al., Nature Protocols 2007 (doi:10.1038/nprot.2007.303).2026-06-01T00:00:00Z2026-06-23T12:53:42.382Z2025-09-29T13:41:43.469ZS-VHPS18