INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY
WORLD HEALTH ORGANIZATION
TOXICOLOGICAL EVALUATION OF SOME
FOOD COLOURS, ENZYMES, FLAVOUR
ENHANCERS, THICKENING AGENTS, AND
CERTAIN FOOD ADDITIVES
WHO FOOD ADDITIVES SERIES 6
The evaluations contained in this publication were prepared by the
Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
4-13 June 19741
World Health Organization Geneva 1975
1 Eighteenth Report of the Joint FAO/WHO Expert Committee on
Food Additives, Wld Hlth Org. techn. Rep. Ser., 1974, No. 557.
FAO Nutrition Meetings Report Series, 1974, No. 54.
This compound has been evaluated for acceptable daily intake by
the Joint FAO/WHO Expert Committee on Food Additives (see Annex 1,
Refs Nos 10 and 20) in 1966 and 1969.
Since the previous evaluation additional data have become
available and are summarized and discussed in the following monograph.
The previously published monographs have been expanded and are
reproduced in their entirety below.
No information available.
Special studies on mutagenicity
This colour was tested for mutagenic effect in a concentration of
0.5 and 1.0 g/100 ml in cultures of Escherichia coli. No mutagenic
effect was found (Lück & Rickerl, 1960).
Special studies on sensitizing effects
In guinea-pigs it was found that this colour had no sensitization
activity (Bär & Griepentrog, 1960). Cats received daily doses of
0.1 g/kg colour for seven days. No increase in Heinz bodies in the
blood of the test animals was noted (Oettel et al., 1965).
Special studies on teratogenicity
Groups of 20-24 pregnant Long-Evans rats received by gavage from
days 6 to 15 of gestation 0, 15, 50 or 150 mg/kg/day. Three control
groups were used and a positive control group of 22 rats was given
30 mg Trypan Blue/kg/day s.c. from days 7 to 9. Rats were sacrificed
at day 20. No signs of maternal or fetal toxicity or anomalies were
seen attributable to the colour. Trypan Blue produced the expected
abnormalities (Anonymous, 1972a).
Groups of 15 pregnant rabbits received by gavage from day
6 to 18 of gestation 0, 15, 50 and 150 mg/kg/day. Three control
groups were used and a positive control group of 15 rabbits was given
150 mg/kg/day thalidomide. Animals were sacrificed on day 29. No
significant maternal or fetal abnormalities due to the colour were
noted. Thalidomide produced the expected abnormalities (Anonymous,
Animal Route mg/kg bw Reference
Rat Oral 2 000 Lu & Lavallee, 1964
Groups of five male and five female rats were fed diets
containing 0, 0.25%, 0.5%, 1.0%, 2.0% and 5.0% for 90 days. No effect
on body weight, food intake, blood cell counts and organ weights was
observed (Hansen et al., 1960).
Ten male and 10 female rats were given a total of 55 subcutaneous
injections of 1 ml of 2% aqueous solution over a period of seven
months, then observed until death. No local tumours developed and
total tumour incidence was less than in control groups given similar
injections of glucose or salt solution (Oettel et al., 1965).
Groups of three male and three female dogs were fed diets
containing 0.03 and 0.2% of the colour for two years. The control
group consisted of 10 animals of each sex. No colour induced effects
were noted in terms of body weight, food consumption, gross and
microscopic pathology (Anonymous, 1967b).
Groups of 20 male and 20 female rats or more were fed diets
containing 0 and 1% of the colour for two years. A similar test group
was formed from the first filial generation and was fed at 1% level
for a similar period of time. No effect of the diet was noted in the
test groups and gross and microscopic examination of the animals
disclosed no charges attributable to the test diet. There was no
significant difference in tumour incidence between the groups (Oettel
et al., 1965).
Groups of 25 male and 25 female rats were fed diets containing
0.0, 0.2 and 0.1% of the colour for up to two years. No colour induced
effects were seen in terms of body weight, food intake, survival,
haematology, urinalyses, organ weights, gross and microscopic
pathology (Anonymous, 1967b).
Twenty rats, half males and half females, received twice weekly
s.c. 1 ml of a 2% aqueous solution into the same site. A total of 55
injections was given for seven months and animals observed for 32
months. Three groups of 20 rats acted as controls. No significant
effects on behaviour, growth, mortality, microscopic appearance of
principal organs was noted. No tumours appeared at the site of
injection of test animals but one sarcoma at the injection site and
two tumours of ovary and uterus appeared in controls (Oettel et al.,
There is no biochemical information but an adequate long-term
study in rats is available. At the previous evaluation a large safety
factor was used but since there are now available additional studies
in non-rodent species, this colour may be evaluated on a basis similar
to other food colours. Multigeneration studies are in progress but
embryotoxicity including teratology has been studied in two species.
Level causing no toxicological effect
Rat: 0.1% (= 1000 ppm) in the diet equivalent to 50 mg/kg bw.
Estimate of acceptable daily intake for man
0-0.5 mg/kg bw*
FURTHER WORK OR INFORMATION
Required by June 1978
Metabolic studies in several species preferably including man.
Adequate long-term study in another species. Results of multi-
Anonymous (1967a) Unpublished report submitted to WHO by Hazelton
Anonymous (1967b) Unpublished report submitted to WHO by Hazelton
Anonymous (1972a) Unpublished report submitted to WHO by Biodynamics
Anonymous (1972b) Unpublished report submitted to WHO by Biodynamics
Bär, F. & Griepentrog, F. (1960) Med. u. Ernähr., 1, 99
BIBRA (1973) Personal communication
Hansen, W. H., Wilson, D.C. & Fitzhugh, O. C. (1960) Fed. Proc., 19,
Lu, F. C. & Lavallee, A. (1964) Canad. pharm. J., 97, 30
Lück, H. & Rickerl, E. (1960) Z. Lebensm.-Untersuch., 112, 157
Oettel, H. et al. (1965) Arch. für Toxikol., 21, 9