Abstract Although, the current market of polymer-based food packaging that use engineered nanoparticles is increasing, concern over toxicity due to their possible migration from polymer packaging into foodstuff affects consumer acceptance which has been recognized as a key factor to negotiate market opportunities successfully. In this context, the aim of this work was to evaluate the performance of films based on α-chitosan (CS)/titanium dioxide (TiO2) nano- and micro- sized complexes in terms of (i) migration according to European Normative 1130-1:2004, (iii) cytotoxicity and (iii) antioxidant activity by means of DPPH tests, as active films for foodstuff. To the best of the authors’ knowledge, this is the first work assessing the migration of TiO2 from CS/TiO2 nano- and micro- sized complexes in accordance with EU Recommendation. With regard to the specific migration tests, which were quantified using acid digestion followed by inductively coupled plasma-mass spectrometry (ICP-MS) detection of titanium, the results revealed that titanium can migrate from chitosan matrix after incubation in different food simulants over 10 days at 40 °C or 10 days at 5 °C, but only in its ionic form, and no evidence of TiO2-particle migration was found. The amount of migrated titanium was in a negligible amount (