INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY WORLD HEALTH ORGANIZATION SUMMARY OF TOXICOLOGICAL DATA OF CERTAIN FOOD ADDITIVES WHO FOOD ADDITIVES SERIES NO. 12 The data contained in this document were examined by the Joint FAO/WHO Expert Committee on Food Additives* Geneva, 18-27 April 1977 Food and Agriculture Organization of the United Nations World Health Organization * Twenty-first Report of the Joint FAO/WHO Expert Committee on Food Additives, Geneva, 1977, WHO Technical Report Series No. 617 30 BLACK 7984 EVALUATION FOR ACCEPTABLE DAILY INTAKE BIOLOGICAL DATA BIOCHEMICAL ASPECTS The dye is poorly absorbed from the gastrointestinal tract after oral administration to rats. Up to 5 g/kg body weight leaves the rat carcass unstained but faeces are black. When given parenterally to mice the whole animal is coloured intensely, this being fully reversible after 12 hours. After i.v. injection of 50 mg/kg body weight into dogs up to 10% is excreted unchanged in the urine some 13% appears as a coloured metabolite (monoazo naphthyl-amino disulfonic acid) while 80% are not accounted for. Only traces of p-sulfanilic acid appear in the urine. Rapid reductive splitting of the azo linkages between the two naphthalene rings occurs (Hecht, 1960a). Acute toxicity Animal Route LD50 mg/kg Reference body weight Mouse intravenous 1 000 mg Hecht, 1960a Rat oral 5 000 mg Hecht, 1960a Rats given 1.5 g/kg body weight orally for 19 days showed no Heinz bodies (Hecht, 1960a). Cats given 100 mg/kg body weight/day for eight days and 5 mg/kg body weight/day for 31 days showed no increase in Heinz bodies (Hecht, 1960a). Special tests Guinea-pig When given s.c. 10 doses of 1 ml of a 0.05-0.1% isotonic saline solution of the colour for a period of 21 days no intolerance was noted. A challenge test for sensitization after 14 days was negative (Hecht, 196Oa). Rabbit Repeated intracutaneous injections of 0.1 ml of a 5% aqueous solution over a few days produced no skin abnormalities (Hecht, 196Oa). Short-term studies Rat The dye was fed to 10 rats for 200 days at a level of 0.1% in the diet. The animals grew normally and no toxic effects were noted (Hecht, 1960a). Long-term studies Rat Two groups of 10 male rats were given 0 or 0.5% of the dye in their drinking water (0.038 g/kg body weight) for 543 days. After this period animals continued on normal diet for up to 822 days. The total intake of colour was approximately 53 g/animal. No abnormalities as regards growth, survival or histopathology were seen and no tumours were found (Hecht, 1960a). In another experiment two groups of 25 male and 25 female rats were given 0 or 0.5% of the colour (average 600 mg/kg body weight) in their drinking water for 337 days and observed for up to 1184 days. Controls were observed up to 1013 days. After 337 days the animals received normal drinking water. Mortality of males only was greater in the test group after 24 months compared with male controls. At 30 months six test females and one control female survived. Nine mammary fibro-adenomas of the breast and one ovarian tumour occurred in female test animals, one sarcoma and five breast fibroadenomas in control females. No extensive pathology was done (Hecht, 1960b). One group of 10 male rats was given twice a week 0.5 ml of a 1% solution (5 mg) s.c. After 105 injections (365 days) the administration was stopped and the last animal died on the six hundred and twenty-seventh day. Average survival was 600 days. No subcutaneous tumours were found (Hecht, 1960). In another experiment a group of 25 male rats were given s.c. 0.5 ml of 2% solution (equiv. 10 mg) for 72 administrations (280 days). The last animal died on the one thousand and twenty-seventh day (average 525 days). No subcutaneous or other tumours were found (Hecht, 1960a). A more recent study (Gangolli et al., 1972), in which Black 7984 was compared with other dye-stuffs for protein binding and their ability to produce subcutaneous sarcomas, showed that Black 7984 produced very mild, self-limiting local reactions although its ability to bind to serum proteins was high. This work supported the conclusion that food colourings which can firmly bind to proteins influence the rate of absorption from the site of subcutaneous injection which leads to the eventual production of local sarcomas. Reproduction studies Rat Four females and one male from each group of the second rat study were used as parent generation to breed an F1 generation after eight months on 0.5% Black 7984 in their drinking water. The F1 pups were kept for 110 days, after weaning on 0.5% dye in their drinking water and killed after 146 days. From 19 F1 females an F2 generation was bred. Twenty-three F2 females were kept on 0.5% dye in their drinking water after weaning for 360 days and then continued on normal drinking water up to 800 days. A similar group of 18 F2 male and 25 F2 female controls were observed for a similar period of 28 months. No abnormalities were noted regarding growth, reproduction or any other effects (Hecht, 1960b). REFERENCES Gangolli, S. D., Grasso, P., Goldberg, L. and Hooson, J. (1972) Protein binding by food colorings in relation to the production of subcutaneous sarcoma, Fd. Cosmet. Toxicol, 10, 449-462 Hecht, G. (1960a) quoted in R. Truhaut (1962), Estratto dei Rendiconti dell' Instituto Superiore di Sanita, XXV, 796-920 (Quotation in text only) Hecht, G. (1960b) Private communication to WHO. Typewritten unpublished document from Forbenfebrik Bayer AG. Paper transmitted to the Subcommittee on the Health Control of Foodstuffs of the Council of Europe
See Also: Toxicological Abbreviations BLACK 7984 (JECFA Evaluation)