INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY WORLD HEALTH ORGANIZATION TOXICOLOGICAL EVALUATION OF SOME FOOD COLOURS, EMULSIFIERS, STABILIZERS, ANTI-CAKING AGENTS AND CERTAIN OTHER SUBSTANCES FAO Nutrition Meetings Report Series No. 46A WHO/FOOD ADD/70.36 The content of this document is the result of the deliberations of the Joint FAO/WHO Expert Committee on Food Additives which met in Rome, 27 May - 4 June 19691 Food and Agriculture Organization of the United Nations World Health Organization 1 Thirteenth report of the Joint FAO/WHO Expert Committee on Food Additives, FAO Nutrition Meetings Report Series, in press; Wld Hlth Org. techn. Rep. Ser., in press. FAST GREEN FCF Biological Data Biochemical aspects Rats and dogs were given orally 200 mg of the colour. In the rats the urine and faeces were collected for 36 hours. In the dogs a bile fistula was made for bile analysis. Almost all (>90 per cent.) the administered colour was excreted unchanged in the faeces of the rats. No colour was found in the urine. In the bile of the dogs the amount of the colour never exceeded five per cent. of the given dose. After feeding, the colour was found in the bile of rats and rabbits, but not in their urine. It was concluded that the quantity found in the bile provides a reasonable estimate of the amount absorbed from the gastrointestinal tract (Hoss & Fitzhugh, 1955, 1953 and 1954). Acute toxicity Animal Route LD50 Reference per kg body-weight rat oral >2.0 g Lu and Lavallée, 1964 Dog. The colour given in single 200 mg doses to dogs did not produce catharsis (Radomski et al., 1956). Short-term studies Rat. Two groups of 16 female rats (control group of 10 rats) were given subcutaneous injections of 0.5 ml of a three per cent. and six per cent. solution. (The rats received with each injection respectively 15 and 30 mg). The used colour was certified as 92 per cent. pure and was supplied as the disodium sulphonate salt. The 10 control rats were given distilled water injections. At first, injections of six per cent. were given three times a week; after 17 weeks it became necessary to reduce the dose to three per cent. Thereafter, both groups were given injections of three per cent. twice weekly for nine weeks. The rest of the time, 22 weeks, both groups were injected usually once a week, occasionally two injections were tolerated. Growth inhibition was found. Thirteen out of 16 animals receiving six per cent. of the colour had fibrosarcomas. The animals given three per cent. showed also fibrosarcomas (10 out of 12). The controls did not show neoplastic tissue at the site of injection (Hesselbach & O'Gara, 1960). Subcutaneous injection of 1 ml of a 0.8 per cent. solution twice weekly produced histological changes suggestive of subsequent sarcoma formation unassociated with chemical carcinogenic potential (Grasso & Goldberg, 1966). Dog. Dog feeding studies were completed. Four beagles per group, equally divided by sex, were fed at 0, 1.0 and 2.0 per cent. for two years. Histopathology attributable to the colour was limited to green blobs of pigment in the renal cortical tubular epithelial cytoplasm of a male dog on a high dose level; a female dog on a high dose level showed slight interstitial nephritis and slight bone marrow hyperplasia (Hansen et al., 1966). Long-term studies Mouse. Chronic mouse feeding studies were conducted; two groups of 100 mice each were fed for two years at 1.0 and 2.0 per cent. and 200 mice served as controls. Microscopic examination revealed no lesions that were attributed to the feeding of the colour (Hansen et al., 1966) Rat. Groups of 50 litter-mated weanling, Osborn-Mendel rats, evenly divided by sex, were fed the colour at 0, 0.5, 1.0, 2.0 and 5.0 per cent. for two years. No effects on growth or mortality were observed. Microscopic examination revealed no lesions that were attributed to the feeding of the colour (Hansen et al., 1966) The colour was fed at a level of four per cent. in the diet to five male and five female rats for periods from 18 to 20 months. This procedure resulted in gross staining of the forestomach, glandular stomach, small intestine, and colon. Granular deposits were noted in the stomach. No tumours were observed (Willheim & Ivy, 1953). Eighteen rats of both sexes were injected this colour subcutaneously for 94 to 99 weeks. In general 1 ml of a two per cent. or three per cent. solution was injected weekly. Subcutaneous fibrosarcomas appeared at the site of injection in 15 of the animals (Nelson & Hagan, 1953; Hansen et al., 1966). Comments: The production of a high percentage of local sarcomata, at the sites of subcutaneous injections in rats has led in the past to considerable discussion and consequently to extensive studies on this colour. The production of these sarcomata is considered to be related to the physico-chemical properties of the colour and the special conditions of the experiment and does not constitute evidence of carcinogenicity by the oral route. Biochemical studies have shown that the colour is poorly absorbed and is almost completely excreted in the faeces after parenteral administration. Extensive long-term studies in two species are available, Biochemical studies on the metabolism using modem techniques are desirable. EVALUATION Level causing no toxicological effect Rat. Five per cent. (= 50 000 ppm) in the diet equivalent to 2500 mg/kg body weight/day Estimate of acceptable daily intake for man: mg/kg body weight Unconditional acceptance 0.-12.5 REFERENCES Grasso, P. & Goldberg, L. (1966) Fd. Cosmet. Toxicol., 4, 269 Hansen, W. H., Long, E. L., Davis, K. J., Nelson, A. A. & Fitzhugh, O. G. (1966) Fd. Cosmet. Toxicol., 4, 389 Hess, S. M. & Fitzhugh, O. G. (1953) Fed. Proc., 12, 330 Hess, S. M. & Fitzhugh, O. G. (1954) Fed. Proc., 13, 365 Hess, S. M. & Fitzhugh, O. G. (1955) J. Pharmacol. exp. Ther., 114, 38 Hesselbach, M. L. & O'Gara, R. W. (1960) J. Nat. Cancer Inst., 24, 769 Lu, F. C. & Lavallée, A. (1964) Canad. pharm, J., 97, 30 Nelson, A. A. & Hagan, E. C. (1953) Fed. Proc., 12, 397 Radomski. J. L. & Deichmann, Wm. B. (1956) J. Pharmacol exp. Ther., 118, 322 Willheim, R. & Ivy, A. C. (1953) Gastroenterology 23, 1
See Also: Toxicological Abbreviations Fast Green FCF (WHO Food Additives Series 16) Fast Green FCF (WHO Food Additives Series 20) Fast Green FCF (WHO Food Additives Series 21) FAST GREEN FCF (JECFA Evaluation) Fast Green FCF (IARC Summary & Evaluation, Volume 16, 1978)