Tire and Road Wear Particles (TRWP) account for an important part of the anthropogenic particles released into the environment. There are scientific knowledge gaps as to the potential bio accessibility of chemicals associated with TRWP to aquatic organisms. This study aimed to investigate the solubilization of five tire-associated chemicals into fish gut using an in vitro digestion model (Oncorhynchus mykiss). Our results show that the targeted compounds were partly and rapidly solubilized into simulated fluids (SF) present in the gastrointestinal tract within a typical gut transit time for fish (3h in SFGASTRIC and 24h in SFINTESTINAL). The effects of food co-ingestion on the solubilization of tire-associated chemicals was compound-specific and either lowered or stimulated their solubilization into the gut fluids. Therefore, the uptake of the tire associated chemicals by the epithelial cells and related toxicity to fish need to be investigated.