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.
BRILLIANT BLACK BN
Biological Data
Biochemical aspects
At concentrations of 2-400 mg/litre the colour inhibits pepsin but not
lipase (Diemair & Häusser, 1951) and at 12.5 mg/1itre it inhibits
trypsin inconsistently (Diemair & Boeckhaff, 1953). Intravenous
injection into rabbits and dogs of 50 mg/kg produced only small
amounts in the urine (Hecht, 1960). Heated in the presence of reducing
sugars the colour is partially decomposed and gives an orange
derivative, isolated by paper chromatography, which is the disodium
salt of 4'(4-sulfo-1-phenylazo) 1'amino 7'sulfonaphtalene (Saent
Lascano Ruiz, Laroche, 1960).
Acute toxicity
Animal Route LD50 Reference
mg/kg body weight
Oral >5 000 DFG, 1957
>2 000 Gaunt et al., 1967
I.P. 500-1 000 Gaunt et al., 1967
Oral >5 000 Gaunt et al, 1967
Rat I.P. 1 100 Gaunt at al., 1967
>2 000 DFG, 1957
I.V. 2 500 DFG, 1957
approx.
Five rats were given 1.5 g/kg body weight orally for 22 days. No Heinz
bodies were found (DFG, 1957). In an experiment with guinea-pigs it
was found that this colour had no sensitization activity (Bar &
Griepentrog, 1960). A cat fed 0.1 g/kg body weight per day for seven
days developed no Heinz bodies (DFG, 1957).
Special studies
The colour was tested for mutagenic effect in a concentration of 0.5
g/100 ml in cultures of Escherichia coli. No mutagenic effect was
found (Lück & Richerl, 1960).
Short-term studies
Rat: Groups of 16 male and 16 female weanling rats were fed diets
containing 0, 0.3 per cent., 1.0 per cent. and 3.0 per cent. colour
for 90 days. Growth retardation associated with diminished food intake
was evident only in males at the three per cent. level. This was shown
by a paired feeding test. Haematological examination, liver and kidney
function tests were normal. Organ weight of testes and kidneys
increased in males at the three per cent. level only. No untoward
histopathological findings were seen (Gaunt et al., 1967).
Long-term studies
Fifty rats were given 0.5 per cent. of the colour in drinking water
for 337 days. The tumours found at 740 days were of the same
distribution as in 50 controls. Another 10 rats received the colour
for 543 days and were observed for 822 days. No tumours were noted.
Another group of 15 rats were fed the colour at 0.5 per cent. in the
drinking water for 360 days and developed no tumours by 800 days.
Fifty animals of the second generation continued with 0.5 per cent. in
the drinking water and had developed no tumours after 425 days (Hecht,
1960). Ten rats were fed the colour at 0.1 per cent. of the diet for
410 days and were observed for 761 days. One animal died prematurely.
No tumours were observed (Hecht & Wingler, 1952; DFG, 1957). Another
group of 10 rats were given twice weekly subcutaneous injections of a
one per cent. solution of the colour for 365 days. Animals were
observed for 816 days. Two rats died but no tumours were observed
(DFG, 1957). Another group of 25 rats received twice weekly
subcutaneous injections of a two per cent. solution of the colour for
280 days. No tumours had appeared by 525 days (Hecht, 1960).
Comments
Little is known of the metabolism and only a long-term study on tumour
incidence in one species is available. A two-year study in a
non-rodent mammalian species is also required. Parenteral
administration does not appear to induce any local neoplastic changes.
EVALUATION
Not possible on the data available.
REFERENCES
Bär, F. & Griepentrog, F. (1960) Med. u. Ernâhr., 1, 99
Deutsche Forschungsgemeinschaft (1957) Fabstoff Komission, Mitteilung,
6
Diemair, W. & Häusser, H. (1951) Z. Lebensmitt.-Untersuch., 92, 165
Diemair, W. & Boekhoff, K. (1953) Z. Analyt. Chem., 139, 35
Gaunt, I. F. et al. (1967) Fd. Cosmet. Toxicol., 5, 171
Hecht, G. & Wingler, A. (1952) Arzneimittel-Forsch., 2, 192
Hecht, G. (1960) Unpublished report to Farbenfabriken Bayer, dated 1
December 1960
Lück, H. & Richerl E. (1960) Z. Lensmitt.-Untersuch., 112, 157
Saenz Lascano Ruiz, I. & Laroche, C. (1960) Ann. Fals. Exp. Chim.,
53, 581