Bladder tumour signal in rats
An oral carcinogenicity study in rats found an increased number of bladder epithelial changes and a small number of bladder tumours in the highest-dose male group compared with controls. The results were judged equivocal by the JECFA committee at the time, partly because the numbers were small and the biological significance was unclear. A later histological re-examination found only three proliferative bladder lesions in high-dose males, leading JECFA to conclude there was no clear neoplastic effect.
An oral rat study showed an increased incidence of urothelial hyperplasia in treated males and a small number of bladder transitional-cell neoplasms in the highest-dose group. JECFA initially set only a temporary ADI pending full histological review.
1984AnimalJECFA Monograph 602, WHO Food Additives Series 20 (Fast Green FCF)
On histological re-examination, only three proliferative bladder lesions were found in high-dose male rats, and JECFA concluded there was no clear indication of a neoplastic effect on the urinary bladder. A permanent ADI of 0-25 mg/kg body weight was then set.
1986AnimalJECFA Monograph 614, WHO Food Additives Series 21 (Fast Green FCF)
Injection-site sarcomas in rats
When Fast Green FCF was injected repeatedly under the skin of rats in laboratory studies, fibrosarcomas developed at the injection sites. IARC reviewed this evidence in 1978 (Volume 16) and re-evaluated it in Supplement 7 (1987), assigning IARC Group 3 on both occasions, meaning the evidence was insufficient to classify the substance as a human carcinogen. The JECFA committee separately concluded that injection-site tumours do not constitute evidence of carcinogenicity by the oral route, the route relevant to food.
Fast Green FCF was carcinogenic in rats, producing sarcomas at the site of repeated subcutaneous injections. No human epidemiological data were available to the Working Group. IARC assigned Group 3: not classifiable as to carcinogenicity in humans. This classification was re-confirmed in Supplement 7 (1987).
1978AnimalIARC Monographs Volume 16 (1978); Supplement 7 (1987) - confirmed via IARC/inchem.org summary
Follicular and neural effects in zebrafish
A 2025 laboratory study exposed zebrafish to Fast Green FCF at concentrations of 0.1%, 0.3% and 0.5% over a prolonged period. Chronic exposure arrested follicular maturation, leaving immature follicles, and reduced acetylcholinesterase activity in the nervous system while increasing markers of oxidative stress. The researchers concluded the dye showed follicular and neural toxicity in this model. Zebrafish studies are an early-stage screening tool and cannot be directly translated to human dietary exposure levels.
Chronic exposure of zebrafish to Fast Green FCF arrested follicular maturation, reduced acetylcholinesterase activity, and increased malondialdehyde levels, indicating oxidative stress and effects on nervous system signalling and reproductive development.
2025Lab + animalSudhakaran G et al., Toxicology Letters, 2025, vol. 413, article 111726
JECFA 2017 re-evaluation
At its 84th meeting in 2017, JECFA reviewed newly available data on Fast Green FCF and reconfirmed the ADI of 0-25 mg/kg body weight. The committee concluded that dietary exposures in all population groups, including the highest adolescent estimate of around 12 mg/kg body weight per day, remain below the ADI and do not present a health concern. The committee noted this ADI applies to countries where the dye is permitted, such as the United States.
JECFA confirmed an ADI of 0-25 mg/kg body weight for Fast Green FCF. New data gave no reason to revise the earlier ADI, and estimated dietary exposures, including the highest adolescent estimate of 12 mg/kg body weight per day, remained below the ADI.
2017Regulatory reviewJECFA 84th Meeting (2017), reconfirmation confirmed via WHO JECFA database (apps.who.int/food-additives-contaminants-jecfa-database); full monograph in WHO Food Additives Series 75 (2019)