FAST GREEN FCF
Explanation
Fast Green FCF was evaluated by JECFA in 1969 (FAO, 1970) and was
allocated an ADI of 0-12.5 mg/kg bw. Since the previous review, new
data have become available and are included in this monograph.
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 the administered colour
was excreted unchanged in the faeces of rats. No colour was found in
the urine. In the bile of the dogs, the amount of colour never
exceeded 5% 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 (Hess & Fitzhugh,
1953, 1954 and 1955). Following i.v. injection in rats, over 90% of
the colour was excreted in the bile within four hours (Iga et al.,
1971).
Fast Green FCF was found to have a high binding affinity for
plasma protein (Gangolli et al., 1967 and 1972; Iga et al., 1971).
TOXICOLOGICAL STUDIES
Special studies on carcinogenicity (see also Long-term studies)
Rat
Eighteen weanling Osborne-Mendel rats of both sexes received
weekly s.c. injections of approximately 30 mg (1 ml of a 3% aqueous
solution) of the colour for 94-99 weeks. Subcutaneous fibrosarcomas
appeared at the site of injection in 15 animals (Nelson & Hagan, 1953;
Hansen et al., 1966).
Two groups of 16 female rats (control group of 10 rats) were
given subcutaneous injections of 0.5 ml of a 3% and 6% solution (the
rats received with each injection respectively 15 and 30 mg). The used
colour was certified as 92% pure and was supplied as the disodium
sulfonate salt. The 10 control rats were given distilled water
injections. At first, injections of 6% were given three times a week;
after 17 weeks it became necessary to reduce the dose to 3%.
Thereafter, both groups were given injections of 3% 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 6%
of the colour had fibrosarcomas. The animals given 3% 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% solution twice weekly
produced histological changes suggestive of subsequent sarcoma
formation unassociated with chemical carcinogenic potential (Grasso &
Golberg, 1966).
No tumours were produced in 11 hamsters injected with 1 mg of the
dye in 0.1 ml water (Price et al., 1978).
Special studies on mutagenicity
Fast Green FCF was non-mutagenic in the Salmonella/microsome
assay (Brown et al., 1978) and negative results were also obtained in
bacterial DNA repair tests (Kada et al., 1972; Rosenkranz & Leifer,
1980). The colour was inactive in a gene conversion assay in diploid
yeast (Sankaranarayanan & Murthy, 1979).
In one of two experiments, the colour-induced cell transformation
in cultured Fisher rat embryo cells at a concentration of 1 µg/ml
(Price et al., 1978) and chromosome damage was reported in an in
vitro test using Chinese hamster ovary cells (Au & Hsu, 1979).
Acute toxicity
LD50
Animal Route per mg/kg bw Reference
Rat Oral > 2000 Lu & Lavallee, 1964
Dog
The colour given in single 200 mg doses to dogs did not produce
catharsis (Radomski et al., 1956).
Short-term studies
Rat
Groups of 50 weanling Osborne-Mendel rats, evenly divided by sex,
were fed diets containing 0, 0.5, 1.0, 2.0 or 5.0% colour 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 dietary level of 4.0% 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).
Dog
Four beagles per group, equally divided by sex, were fed at 0,
1.0 and 2.0% 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
Groups of 50 male and 50 female C3HeB/FeJ mice were fed diets
containing 1.0 or 2.0% colour for two years and 100 mice of each sex
served as controls. After 78 weeks, 56 controls and 27 animals in the
1.0%, 17 animals in the 2.0%, treatment groups still survived.
Microscopic examination revealed no lesions that were attributed to
feeding of the colour (Hansen et al., 1966).
Rat
Groups of 50 weanling Osborne-Mendel rats, evenly divided by
sex, were fed diets containing 0, 0.5, 1.0, 2.0 or 5.0% colour 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 dietary level of 4.0% 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).
OBSERVATIONS IN MAN
No data available.
Comments
The production of a high percentage of local sarcomata at the
site of subcutaneous injection in rats is considered to be related to
the physicochemical properties of the colour and the special
conditions of the experiments, and does not constitute evidence of
carcinogenicity by the oral route. However, in reviewing the
carcinogenicity of Fast Green FCF, an IARC Working Group emphasized
the inadequacy of the long-term feeding studies in mice and rats
(IARC, 1978); in the rat study, only 12 animals fed 5% of the dye were
subjected to detailed microscopical examination.
Biochemical studies have shown that the colour is poorly absorbed
and is almost completely excreted in faeces after parenteral
administration.
No data were available on reproductive toxicology and teratology.
EVALUATION
Level causing no toxicological effect
Rat: 5% in the diet equivalent to 2500 mg/kg bw.
Estimate of temporary acceptable daily intake for man
0-12.5 mg/kg bw.
FURTHER WORK OR INFORMATION
Required by 1985
Adequate long-term feeding studies in the rat and a
multigeneration reproduction/teratology study.
REFERENCES
Au, W. & Hsu, T. C. (1979) Studies on clastogenic effects of biologic
stains and dyes, Environmental Mutagenesis, 1, 27
Brown, J.P., Roehm, G. W. & Brown, R. J. (1978) Mutagenicity testing
of certified food colours and related azo, xanthene and
triphenylmethane dyes with the Salmonella/microtome system,
Mutation Res., 56, 249-271
FAO (1970) Toxicological evaluation of some food colours, emulsifiers,
stabilizers, anti-caking agents and certain other substances,
FAO Nutr. Mtgs. Rep. Series No. 46A
Gangolli, S. D., Grasso, P. & Golberg, L. (1967) Physical factors
determining the early local tissue reactions produced by food
colourings and other compounds injected subcutaneously, Fd.
Cosmet. Toxicol., 5, 601-621
Gangolli, S. D., et al, (1972) Protein binding by food colourings in
relation to the production of subcutaneous sarcoma, Fd. Cosmet.
Toxicol., 10, 449-462
Grasso, P. & Golberg, L. (1966b) Subcutaneous sarcoma as an index of
carcinogenic potency, Fd. Cosmet. Toxicol., 4, 297
Hansen, W. H. et al. (1966) Chronic toxicity of three food colourings:
guinea green B, light green SF yellowish, and fast green FCF in
rats, dogs and mice, Fd. Cosmet. Toxicol., 4, 389-410
Hess, S. M. & Fitzhugh, O. G. (1953) Metabolism of coal-tar dyes. I.
Triphenylmethane dyes (Abstract No. 1090), Fed. Proc., 12,
330-331
Hess, S. M. & Fitzhugh, O. G. (1954) Metabolism of coal-tar colours.
II. Bile studies (Abstract No. 1201), Fed. Proc., 13, 365
Hess, S. M. & Fitzhugh, O. G. (1955) Absorption and excretion of
certain triphenylmethane colours in rats and dogs,
J. Pharmacol. Exp. Ther., 114, 38-42
Hesselbach, M. L & O'Gara, R. W. (1960) Fast green and light green
induced tumours: induction, morphology and effect on host,
J. nat. Cancer Inst., 24, 769-793
IARC (1978) IARC Monographs on the Evaluation of the Carcinogenesis
Risk of Chemicals to Man: some aromatic amines and related nitro
compounds - hair dyes, colouring agents and miscellaneous
industrial chemicals. Vol. 16, IARC, Lyon
Iga, T., Awazu, S. & Nogami, H. (1971) Pharmacokinetic study of
biliary excretion. II. Comparison of excretion behaviour in
triphenylmethane dyes, Chem. pharm. Bull., 19, 273-281
Kada, T., Tutikawa, K. & Sadaie, Y. (1972) In vitro and
host-mediated 'rec-Assay' procedures for screening chemical
mutagens and phloxine, a mutagenic red dye detected, Mutation
Res.,16, 165-174
Lu, F. C. & Lavallee, A. (1964) The acute toxicity of some synthetic
colours used in drugs and foods, Canad. pharm. J., 97, 30
Nelson, A. A. & Hagan, E. C. (1953) Production of fibrosarcomas in
rats at site of subcutaneous injection of various food dyes
(Abstract No.1307), Fed. Proc., 12, 397-398
Price, P. J. et al. (1978) In vitro and in vivo indications of the
carcinogenicity and toxicity of food dyes, Int. J. Cancer,
21, 361-367
Radomski, J. L. & Deichman, W. B. (1956) Cathartic action and
metabolism of certain coal tar food dyes, J. Pharmacol. Exp.
Ther., 118, 322-327
Rosenkranz, H. S. & Leifer, Z. (1980) In Chemical Mutagens: Principles
and Methods for their Detection. In: de Serres, F. J. &
Hollaender, A., New York and London, Plenum Press, Vol. 6, p. 109
Sankaranarayanan, M. & Murthy, M. S.S. (1979) Testing of some
permitted food colours for the induction of gene conversion in
diploid yeast, Mutation Res., 67, 309-314
Willheim, R. & Ivy, A. C. (1953) A preliminary study concerning the
possibility of dietary carcinogenesis, Gastroenterology, 23,
1-19
See Also:
Toxicological Abbreviations
Fast green FCF (FAO Nutrition Meetings Report Series 46a)
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)