Food seems to be one of the main ways of animal and human contamination with polycyclic aromatic hydrocarbons (PAHs) and dioxins. In vivo studies showed a blood absorption of these xenobiotics after their ingestion. Our work aimed at studying the in vitro transfer of PAHs and dioxins through intestinal barrier. Caco-2 cells were cultivated on permeable filters to measure transepithelial permeability of 14C labeled 2,3,7,8- tetrachlorodibenzo-p-dioxin, benzo[a]pyrene, pyrene, and phenanthrene, which differed in their physicochemical properties. The results showed that the molecules were able to cross intestinal cell layers. All the molecules were detected associated with cells, even if the dioxin was the less uptaken compound. Phenanthrene appeared in basal media faster, and its level after a 6-h exposure was respectively 1.1, 2, and 7 times higher than pyrene, benzo[a]pyrene, and 2,3,7,8-tetrachlorodibenzo-p-dioxin levels. These findings suggest that intestinal epithelium plays a key role in selective permeability and then in bioavailibility of micropollutants.