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

    In dog it was found that 6-41 per cent. of the administered colour was
    excreted in the faeces. No colour was found in the urine. Incubating
    the colour with a dog's fresh intestinal content destroyed 80 per
    cent. of the colour in one hour (DFG Dye Commission, 1957; Vos et al.,

    Acute toxicity


    Animal   Route            LD50                Reference
                         mg/kg body-weight

    Rat      i.p.             1 000               DFG, 1957

    Short-term studies

    Rat. Ten rats were fed this colour at a level of four per cent. in
    the diet. The animals lived only a few months. Gross staining of the
    glandular stomach and the small intestine, and granular deposits in
    the stomach and small intestine were observed (Willheim & Ivy, 1953).

    Weanling male rats were fed diets containing 1, 2 or 5 per cent. of
    the colour for 16 weeks. The animals with the 2 and 5 per cent. died
    all within the first two weeks of the feeding period. A mortality of
    80 per cent. occurred in the rats fed one per cent. and the growth
    rate of the surviving animals in this group was markedly retarded. The
    inclusion of the colour in the diet caused severe diarrhoea,
    enlargement of the spleen and an anaemia (Hallesy & Doull, 1956).

    Dog. This colour had a significant cathartic effect when 100 to 200
    mg were given to groups of five dogs (Vos et al., 1953)

    Man. This colour induces also a cathartic action in man (80 mg). The
    intact molecule appears to be the active cathartic agent (Radomski &
    Deichmann, 1956).

    Human volunteers who ate candy containing 0.07 per cent. of this
    colour exhibited diarrhoea upon ingestion of one to eight pieces of
    the candy (US FDA, 1955).

    Skin tests with this colour showed no reaction in patients sensitive
    to paraphenylene diamine (Baer et al., 1948).

    Long-term toxicity

    Mouse. Twenty mice were fed 15-20 mg per week of this colour for
    periods up to 409 days five days a week. No tumours were observed
    which could be attributed to the colour (Cook et al., 1940).

    Rat. This colour was fed to 85 rats at a level of 0.1 per cent. in
    the diet. The daily intake varied from 10 to 15 mg for a period of 400
    days. No tumours were observed (DFG, 1957).

    Groups of 24 weanling rats, equally divided by sex, were fed the
    colour at 0, 0.5, 1.0 and 2.0 per cent. in the diet. No rats fed at
    2.0 per cent. survived beyond the fifth week, observations are
    confined to 0, 0.5 and 1.0 per cent. levels. Body-weight and food
    intake were recorded weekly for two years; blood counts were taken
    four times. Gross findings at 1.0 per cent. were marked increase of
    mortality, enlargement of spleens, leukocytosis and anaemia,
    diarrhoea, and growth depression. At 0.5 per cent. kidneys of test
    rats showed more chronic congestion than those of controls and there
    was some splenic enlargement. Microscopically, spleens showed
    uniformly chronic congestion and less often slight hyperplasia and
    increased pigmentation. Kidneys showed nephritis of the type common in
    older rats with no difference among the groups except in incidence
    (Bourke et al., 1956; US FDA, 1963).

    Eighteen young rats were given subcutaneous injections, 20 mg of the
    colour per week (two per cent. aqueous solution) for two years. In six
    cases fibresarcomas were found. The controls did not get tumours
    (Nelson & Davidow, 1967).

    Dog. The colour was given to 14 dogs at four levels in daily
    quantities by capsule ranging from about 0.02 per cent. to 1.0 per
    cent. in the diet. The dogs on the lowest level survived for five
    years without showing any effect. At higher levels (0.2 per cent. or
    more of the diet) effects were variable; some dogs survived only for
    short periods, others showed little or no effect for long periods.
    Pathological changes were generally non-specific; some animals were
    found dead with little to explain the death; others were emaciated and
    showed organ changes chiefly of inanition (Bourke et al., 1956; US
    FDA, 1963).


    None of the long-term studies except the study carried out in dogs
    revealed a satisfactory no-effect level. Further metabolic studies in
    several species, preferably including man, are required as well as
    long-term studies in a rodent species to establish a no-effect level.


    Not possible on the data available.


    Baer, R. L., Leider, M. & Mayer, R. L. (1948) Proc Soc. exp. Biol.,
    67,  489

    Bourke, A. R., Nelson, A. A. & Fitzhugh, O. G. (1956) Fed Proc., 15,

    Cook, J. W., Hewett, C. L., Kennaway, E. L. & Hennaway, N. M. (1940)
    Amer. J. Cancer, 40, 62

    Deutsche Forschungsgemeinschaft, Farbstoff Kommission (1957)
    Mitteilung 6

    Hallesy, D. W. & Doull, J. (1956) J. Pharmacol. exp. Ther.,  116, 26

    Nelson, A. A, & Davidow, B. (1957) Fed. Proc., 16, 367

    Radomski, J. L. & Deichmann, H. B. (1956) J. Pharmacol. exp. Ther.,
    118, 322

    United States Food and Drug Administration (1955) Title 21, Food and
    Drugs, Part 135, Federal Register, 20, 8492

    United States Food and Drug Administration (1963) Unpublished report

    Vos, B. J., Radomski, J. L. & Fuyat, H. N. (1953) Fed. Proc., 12,

    Willheim, R. & Ivy, A. C. (1953) Gastroenterology, 23, 1

    See Also:
       Toxicological Abbreviations