Positive genotoxicity signals in laboratory tests
When red iron oxide (and black iron oxide) were tested in mammalian cell cultures, they caused DNA strand breaks and formation of micronuclei, which are markers of genetic damage. These effects were seen in both nano-sized and larger particles. However, when red iron oxide was given orally to rats, no genotoxic effects were found in blood-forming cells. Because the two sets of results point in opposite directions and the testing database was incomplete, the EFSA Panel concluded it could not determine whether iron oxides pose a genotoxic risk.
Red and black iron oxide, in both nano and micro particle sizes, produced positive results in in vitro genotoxicity assays in mammalian cells, including DNA strand breaks and micronuclei induction.
2015LabEFSA Panel on Food Additives and Nutrient Sources (ANS), Scientific Opinion on the re-evaluation of iron oxides and hydroxides (E 172) as food additives, EFSA Journal 2015;13(12):4317
In vivo oral administration of nano- and microsized red iron oxide did not elicit genotoxic effects in rat haemopoietic cells; however no in vivo data are available for the gastrointestinal tract, which is the site of direct contact.
2015AnimalEFSA Panel on Food Additives and Nutrient Sources (ANS), EFSA Journal 2015;13(12):4317
EFSA concluded a full safety assessment was not possible
When EFSA re-evaluated E172 in 2015, the panel found that the available biological and toxicological data were insufficient to carry out an adequate safety assessment. No new acceptable daily intake was set. The panel called for additional data on genotoxicity, carcinogenicity, and reproductive and developmental toxicity before a firm conclusion could be reached. This is a notable regulatory outcome: for most long-approved additives, EFSA reaches a positive conclusion; for E172, it explicitly could not.
The EFSA ANS Panel concluded that an adequate assessment of the safety of E 172 could not be carried out because a sufficient biological and toxicological database was not available, and recommended additional toxicological data be provided.
2015Regulatory reviewEFSA Panel on Food Additives and Nutrient Sources (ANS), EFSA Journal 2015;13(12):4317
No acceptable daily intake was allocated by EFSA in the 2015 re-evaluation. JECFA had previously set a group ADI of 0 to 0.5mg/kg body weight per day in 1980, but this was based on a much older and thinner dataset.
1980RegulatoryJECFA (Joint FAO/WHO Expert Committee on Food Additives), monograph 1980; EFSA Journal 2015;13(12):4317
Nano-sized particles present in commercial supplies
A 2020 study that characterised seven commercially available E172 samples found that more than 50% of particles in all samples had at least one dimension below 100 nanometres. Current EU and UK specifications for E172 do not require manufacturers to state particle size or the nano fraction. This matters because nano-sized particles can behave differently in the body from larger versions of the same material, and the 2015 EFSA opinion specifically flagged particle size as a data gap that needed to be addressed.
More than 50% of iron oxide particles detected in commercial E172 pigment samples fell in the nanoscale range (at least one external dimension 1 to 100nm), yet current regulatory specifications contain no particle-size requirements.
2020LabVoss et al., Food Chemistry, 2020 (DOI: 10.1016/j.foodchem.2020.127000)
EFSA recommended that particle size and particle size distribution should be included in the specifications for E172, noting this information was absent from the submitted dossier.
2015Regulatory reviewEFSA Panel on Food Additives and Nutrient Sources (ANS), EFSA Journal 2015;13(12):4317
Behaviour in the gut and liver cells at high doses
Studies using simulated digestion show that iron oxide particles pass through the gastrointestinal tract without dissolving and are taken up only minimally into intestinal cells. Liver cells in culture showed greater interaction with iron oxide particles at high experimental concentrations, including changes in mitochondrial activity in some particle types, though no cell death was recorded at doses tested. These findings are from laboratory and cell culture models, not from human dietary exposure studies.
In artificial digestion models, iron oxide food colorant particles (E172) passed through without dissolving; only minor uptake into intestinal (Caco-2) cells was detected and negligible transport across the intestinal barrier.
2024LabBrun et al., Food and Chemical Toxicology, 2024 (DOI: 10.1016/j.fct.2024.114447)
At high experimental iron concentrations (100 to 200 micrograms Fe/mL), some iron oxide particle types impaired mitochondrial membrane potential in human liver (HepaRG) cells and triggered apoptosis signalling, though ATP levels remained stable and no significant cell death was observed.
2021LabStern et al., Archives of Toxicology, 2021 (DOI: 10.1007/s00204-020-02960-7)