Information session: Retrospective review of 90-day toxicity studies for regulatory assessment of food enzymes

Within the context of EU roadmap towards phasing out animal testing for chemical safety assessment, EFSA initiated a self-task mandate to review a collection of 90-day rodent studies that have been submitted by applicants for food enzyme safety assessment. In the course of its work, on 18 May 2026, EFSA has brought together internal and external stakeholders to present the preliminary results of the retrospective review, and to discuss how alternative methods that could ensure scientific robustness of future safety evaluation of food enzymes while reducing reliance on animal testing.

Key preliminary findings

The retrospective review of 258 90-day rodent studies found no safety concern in most of the studies. In the small number of cases where toxicological effects were seen, they were linked either to local (gastric) effects in rodents that are not considered relevant for humans, or to impurities and contamination related to the manufacturing process rather than to the enzyme itself. Overall, the preliminary findings confirm that food enzymes are generally of low toxicological concern, and suggest that routine 90-day animal studies may offer limited additional value.

Stakeholder views

182 stakeholders registered and 114 attended the info-session. External stakeholders from academia, industry and regulatory bodies did not question or challenge the statistical methods and overall methodology that EFSA used to conduct the retrospective review. The stakeholders generally supported the use of alternative methods whenever they can provide robust evidence for safety assessment. Among the alternative approaches discussed were literature reviews, evidence of safe use, genomic and protein sequence analysis, digestibility data, impurity analytics, in vitro toxicity testing methods for targeted checks for risks linked to the enzyme protein, source organism, enzyme production process. Participants also highlighted the need for clear guidance so that these approaches can be applied consistently and confidently in regulatory assessments.

Next steps

The work of the self-task mandate is still on-going. EFSA aims to release the draft document for public consultation in autumn 2026. Another stakeholder engagement is planned during the public consultation, for which an announcement will be made soon.