Perfluorooctane sulfonic acid (PFOS) belongs to a large group of anthropogenic compounds with high persistency named per- and polyfluorinated substances (PFAS). Widespread use from industry to household appliances and food-contact materials contributes to PFAS exposure with food as the primary source. Association studies suggest that vegetables and fibre rich diet may reduce PFOS levels in humans, but experimental data remain limited. Here, we investigated PFOS uptake and wash-out after seven days of PFOS (3 mg/kg/day) in two groups of rats (N = 12 per group) fed diets either high (HF) or low (LF) in soluble dietary fibres. Two control groups (N = 12/group) were fed the same diets without PFOS. Changes in pH and transit time were monitored alongside intestinal and faecal microbiota composition. We quantified systemic and excreted, linear and branched PFOS. Results revealed significantly lower pH and faster intestinal transit in the HF groups. Importantly, HF rats had lower serum PFOS concentrations and higher PFOS concentrations in caecal content and faeces, indicating a more efficient excretion on the fibre rich diet. In both dietary groups, PFOS affected the gut microbiota composition. Our results suggest that a diet rich in soluble dietary fibres accelerates excretion of PFOS and lowers PFOS concentration in serum.