The production of titanium dioxide nanoparticles (TiO2 NPs) for commercial applications has greatly increased over the last years and consequently the potential risk for human health. There is a growing awareness of the need to understand the behavior and influence these nanoparticles exert on the environment. Bioaccumulation serves as a good integrator to assess chemical exposure in aquatic systems and is dependent on factors, such as the exposure routes, diet and the aqueous medium. We analyzed the experimental bioaccumulation capability of ionic titanium and TiO2 NPs by zebrafish (Danio rerio) eleutheroembryos through bioconcentration factors (BCFs), after 48 or 72 h of exposure. The stability of both chemical forms in an aquatic medium was fully characterized for further bioaccumulation studies. Several stabilizing agents (humic acids, soluble starch, polyethylene glycol, Na4P2O7 and Na2HPO4) for anatase and rutile, the two allotrophs of TiO2 NPs, were evaluated to check the evolution of the aggregation process. Around 60% of TiO2 NPs remained disaggregated under simulated environmental conditions with the addition of 50 mg L−1 of humic acids. However, the presence of eleutheroembryos in the exposure medium increased TiO2 NPs aggregation in the experimental tests. The BCFs values obtained in all cases were <100, which classifies ionic titanium and TiO2 NPs as non-bioaccumulative substances, under the REACH regulations.