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.


    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.,

    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).


    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.


    Level causing no toxicological effect

    Rat. Five per cent. (= 50 000 ppm) in the diet equivalent to 2500
    mg/kg body weight/day 

    Estimate of acceptable daily intake for man:

                                               mg/kg body weight

    Unconditional acceptance                         0.-12.5


    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,

    Hesselbach, M. L. & O'Gara, R. W. (1960) J. Nat. Cancer Inst., 24,

    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

    See Also:
       Toxicological Abbreviations
       Fast Green FCF (WHO Food Additives Series 16)
       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)