Dietary uptake is important for trophic transfer of polycyclic aromatic hydrocarbons (PAHs) in the freshwater pelagic ecosystem. In this study, we hypothesized that both the dietary uptake rate and interval significantly influenced its relative contribution to bioaccumulation. We developed a toxicokinetic model framework for the bioaccumulation of deuterated PAHs (PAHs-d10) in aquatic organisms considering different feeding intervals ranging from none for phytoplankton to approximately continuous for zooplankton to discrete for fish and built a simple artificial freshwater pelagic food chain composed of algae Chlorella vulgaris, zooplankton Daphnia magna, and zebrafish. We conducted bioaccumulation experiments and simulations for Daphnia magna and zebrafish under different algal densities based on our model. The results showed that intermittent feeding led to a large fluctuation in the PAH-d10 concentrations in zebrafish compared to a leveled-off pattern in Daphnia magna because of approximately continuous feeding. Trophic dilution of PAHs-d10 occurred in the food chain when there was waterborne-only uptake, but dietary uptake largely mitigated its extent that depended on dietary uptake rates. The assimilation efficiency, dietary uptake rate, and its relative contribution to bioaccumulation of PAHs-d10 in zebrafish were all higher than those in Daphnia magna, suggesting that dietary uptake played a more important role in bioaccumulation of PAHs at higher trophic-level organisms.