The data contained in this document were examined by the
    Joint FAO/WHO Expert Committee on Food Additives*
    Geneva, 18-27 April 1977

    Food and Agriculture Organization of the United Nations
    World Health Organization

    * Twenty-first Report of the Joint FAO/WHO Expert Committee on Food
    Additives, Geneva, 1977, WHO Technical Report Series No. 617





         In dogs it was found that 6 to 41% 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% of the
    colour in one hour (DFG Dye Commission, 1957; Vos et al., 1953).

         The urine of a dog given 5 g Orange I contained a small quantity
    of the unchanged colour together with sulphanilic acid and 4-amino-
    1-naphthol after 48 hours (Sisley and Porcher, 1911). (The unchanged
    colour in the urine could be due to contamination.)

         Oral administration of 100 mg of Orange I to a dog with a bile
    fistule resulted in the recovery of only 0.4% in the bile, compared to
    a 47% recovery after s.c. administration of the dye (Radomski and
    Deichmann, 1956). Low excretion of Orange I in animals not showing
    diarrhoea is due to its degradation by intestinal flora, as
    demonstrable by in vitro tests, rather than to its absorption
    through the intestinal wall (Radomski and Deichmann, 1956).

    Acute toxicity


    Animal    Route          LD50 mg/kg       Reference
                             mg/kg bw
    Rat       i.p.           1 000 mg          Truhaut, 1955

         Concentration of 1% of the dye in the diet caused some deaths,
    spleen enlargement, leucocytosis, anaemia, diarrhoea and growth
    depression in rats; 0.5% of the diet increased the incidence of
    nephritis and enlargement of the spleen with hyperplasia and increased
    pigmentation (Bourke et al., 1956).

    Cathartic effect

         Orange I induces catharsis in the rat at a concentration of
    20 mg/kg body weight. The cathartic action of 200 mg of Orange I was
    reported when administered orally in dogs (Radomski and Deichmann,


         Mutagenicity testing of Orange I gave questionable or variable
    responses following reduction and microsomal activation (Brown et al.,

    Short-term studies


         Ten rats were fed this colour at a level of 4% 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 and Ivy, 1953).

         Weanling male rats were fed diets containing 1, 2 or 5% of the
    colour for 16 weeks. The animals with the 2 and 5% died all within the
    first two weeks of the feeding period. A mortality of 80% occurred in
    the rats fed 1% 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 and Doull, 1956).


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

    Long-term studies


         No liver tumours were observed in a group of 20 mice (sex and
    strain unspecified) which received weekly doses of 15-20 mg Orange I
    for life, given over five days a week in the form of a solution in tap
    water added to 20 g brown bread. The last surviving mouse died
    409 days after the start of treatment; the median lifespan was not
    reported. Autopsies were carried out on 14 mice and sections examined
    for seven mice (Cook et al., 1940). [The short duration of the
    experiment was noted by the working group.]


         This colour was fed to 85 rats at a level of 0.1% 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% in the diet. No rats fed at 2.0%
    survived beyond the fifth week, observations are confined to 0, 0.5
    and 1.0% levels. Body weight and food intake were recorded weekly for
    two years; blood counts were taken four times. Gross findings at 1.0%
    were marked increase of mortality, enlargement of spleens,
    leukocytosis and anaemia, diarrhoea and growth depression. At 0.5%
    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 (2% aqueous solution) for two years. In six cases
    fibrosarcomas were found. The controls did not get tumours (Nelson and
    Davidow, 1957).


         The colour was given to 14 dogs at four levels in daily
    quantities by capsule ranging from about 0.02% to 1.0% in the diet.
    The dogs on the lowest level survived for five years without showing
    any effect. At higher levels (0.2% 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).


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

         Human volunteers who ate candy containing 0.07% 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).


    Baer, R. L., Leider, M. and Mayer, R. L. (1948) Eczematous cross-
    hypersensitivity between p-phenylenediamine and azo dyes certified for
    use in foods, drugs and cosmetics, Proc. Soc. exp. Biol., 67, 489

    Bourke, A. R., Nelson, A. A. and Fitzhugh, O. G. (1956) Chronic
    toxicity of FD and C Orange No. 1, Fed. Proc., 15, 404

    Brown, J.P., Roehm, G. W. and Brown, R. J. (1977) Mutagenicity testing
    of certified food colors and related azo, xanthene and triphenyl-
    methane dyes with the Salmonella/microsome system, 8th Ann. Meeting,
    Environmental Mutagen Society, Colorado Springs, Feb. 1977 (abstract)

    Cook, J. W., Hewett, C. L., Kennaway, E. L. and Kennaway, N. B. (1940)
    Effects produced in the livers of mice by azo-naphthalenes and related
    compounds, Amer. J. Cancer, 40, 62-77

    DFG (Deutsche Forschungsgemeinschaft) (1957) Farbstoff-Kommission,
    Toxikologische Daten von Farbstoffen und ihre Zulassung fur
    Lebensmittel in verschiedenen Ländern, Mitt. 6(2), Wiesbaden,
    Steiner Verlag, p. 25

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

    Nelson, A. A. and Davidow, B. (1957) Injection site fibrosarcoma
    production in rats by food colors., Fed. Proc., 16, 367

    Radomski, J. L. and Deichmann, W. B. (1956) Cathartic action and
    metabolism of certain coal-tar food dyes, J. Pharmacol. exp. Ther.,
    118, 322-237

    Sisley, P. and Porcher, C. (1911) Du sort des matières colorantes dans
    l'organisme animal, C.R. Aced. Sci. (Paris), 152, 1062-1064

    Truhaut, R. (1958) Sur l'utilisation des colorants en thérapeutique et
    les dangers qui peuvent en résulter pour la santé humaine, Chimie
    moderne, 3, 337-350

    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. and Fuyat, H. N. (1953) Fed. Proc.,
    12, 376

    Willheim, R. and Ivy, A. C. (1953) Dietary carcinogenesis,
    Gastroenterology, 23, 1

    See Also:
       Toxicological Abbreviations
       ORANGE I (JECFA Evaluation)
       Orange I (IARC Summary & Evaluation, Volume 8, 1975)