Green #3 FDA report 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 toxicological effect
Rat. Five per cent. (= 50 000 ppm) in the diet equivalent to 25000
mg/kg body weight/day
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
