Corrugated cardboard is a sustainable alternative for frozen food packaging, but concerns remain regarding barrier performance and substance migration. This study assessed overall migration and the potential transfer of cytotoxic substances from a beeswax–chitosan antimicrobial coating intended for frozen Atlantic cod packaging.

Overall migration was evaluated on coated and uncoated cardboard using TENAX in stainless steel migration cells for 10 days at 40 °C, in accordance with EN 1186. Migration values were 0.233 ± 0.047 mg/dm² for uncoated cardboard and 0.567 ± 0.047 mg/dm² for coated samples, remaining well below the legal limit of 10 mg/dm² established by EU Regulation 10/2011.

To verify compliance with EDQM 2021 requirements for active packaging, agar diffusion tests were performed to assess the transfer of cytotoxic or antimicrobial compounds. Coated and uncoated samples were tested alongside antibiotic controls on agar plates inoculated with Bacillus subtilis and incubated for 72 h. No inhibition zones were observed for coated samples, indicating no compound transfer.

These results demonstrate that the bio-coated corrugated cardboard exhibits controlled migration behaviour and meets current European regulatory requirements, supporting its potential application as sustainable packaging for frozen fish products.