The evaluations contained in this publication were prepared by the
    Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
    4-13 June 19741

    World Health Organization     Geneva     1975


    1  Eighteenth Report of the Joint FAO/WHO Expert Committee on
    Food Additives, Wld Hlth Org. techn. Rep. Ser., 1974, No. 557.
    FAO Nutrition Meetings Report Series, 1974, No. 54.



         This compound has been evaluated for acceptable daily intake by
    the Joint FAO/WHO Expert Committee on Food Additives (see Annex 1,
    Refs Nos 10 and 20) in 1964 and 1969.

         Since the previous evaluation additional data have become
    available and are summarized and discussed in the following monograph.
    The previously published monographs have been expanded and are
    reproduced in their entirety below.



         "Ponceau 4 R is reduced in vitro by rat caecal contents
    (Walker, 1968), but Ryan and Wright (1961) showed that rats excreted
    30-45% of an intravenous dose unchanged in the bile within 6 hours.
    The bile was coloured in rats and mice given intraperitoneal doses of
    the dye (Gaunt et al., 1967)."


    Special studies on mutagenicity

         This colour was tested for mutagenic effect in a concentration of
    0.5 g/100 ml in cultures of Escherichia coli. No mutagenic effect
    was found (Lück & Rickerl, 1960).

    Special studies on sensitization

         In experiments with guinea-pigs it was found that this colour had
    no sensitization (Bär & Griepentrog, 1960). A negative test for Heinz
    bodies was obtained after administering this colour in a 5% aqueous
    solution by stomach tube to four cats (DFG, 1957).

    Special studies on teratogenicity

         Groups of unspecified numbers of mice received daily doses of
    7.5, 30 or 100 mg/kg bw orange RN by gavage either from day 0-7 or
    day 6-18 of pregnancy. Litter parameters and implantation showed no
    adverse effects. No dose-related abnormalities were seen in the
    fetuses (Larsson, 1974).

    Acute toxicity

                        LD50                Reference
    Animal    Route     mg/kg bw

    Mouse     Oral      8 000               Gaunt et al., 1967

              i.p.      > 1 750 approx.     Gaunt et al., 1967

    Rat       Oral      >8 000              Gaunt et al., 1967

              i.p.      2 000 approx.       DFG, 1957

    Female    i.p.      2 600               Gaunt et al., 1967

    Male      i.p.      600                 Gaunt et al., 1967

              i.v.      1 000               DFG, 1957

    Short-term studies


         Groups of 16 male and 16 female rats were fed diets containing 
    0, 0.5%, 1% and 2% dye for 90 days. No adverse effects were seen in
    appearance, behaviour, growth, food consumption, haematological
    indices, SGPT and SGOT serum levels except at 2% level, when females
    had slightly increased transaminase values, red cell counts and
    haemoglobin concentration. Renal function tests and organ weights were
    normal. Gross and histopathology showed no difference between the test
    groups (Gaunt et al., 1967).

         Eleven rats were given 1.0% of the colour in their drinking water
    for 216 days and observed for 791 days. Two animals died during the
    experiment, one had a sarcoma of the liver (DFG, 1957).


         Four groups of three large white pigs of each sex were fed 0
    (control), 100, 300 or 900 mg Ponceau 4R/kg bw/day for three months.
    One female from the highest dose level died on day 23, but the death
    was attributed to an enteric infection. No differences between treated
    and control pigs were detected in growth, composition of urine and
    serum, organ weights or histopathology. There was a slight reduction
    in the number of erythrocytes at week 6 in males for 900 mg/kg/day
    (Gaunt et al., 1969).

    Long-term studies


         Groups of 30 mice of each sex (Ash-CS1 strain) were fed on diets
    containing 0.01, 0.05, 0.25 and 1.25% Ponceau 4R for 82 weeks with a
    group of 60 mice of each sex serving as controls. There was evidence
    that the colouring was converted to a yellow metabolite in the
    gastrointestinal tract. The feeding of Ponceau 4R had no adverse
    effect on mortality, body weight gain, organ weight or the incidence
    of tumours. A mild anaemia was present in the first six months of the
    study in mice given 0.25 or 1.25% Ponceau 4R. There was an increased
    incidence of foamy reticulo-endothelial cells in the liver at the
    1.25% level and an increased incidence of glomerulonephrosis at this
    and the 0.25% level.

         This study shows that the colouring has no carcinogenic potential
    in mice when fed at dietary concentrations of up to 1.25%. The no-
    untoward-effect level was 0.05% (Gaunt et al., 1974).


         Ten rats were given 0.2% of the colour in their diet for 417
    days. The total intake was 11 g/animal. Observation extended for 1011
    days. One rat died. No tumours were found (DFG, 1957).

         In a similar experiment the dye was given to 11 rats as a 1%
    solution in the drinking water, the daily dose being 1 g/kg for a
    period of 216 days. Total ingestion amounted to 52 g/animal. Animals
    were observed for 791 days. One rat developed a sarcoma in the liver,
    two others died (DFG, 1957).

         Four groups of 10 male and 10 female rats were given diets
    containing 0, 0.03%, 0.3%, and 3.0% of the colour for 64 weeks. No
    effect was noted on mortality. Females at the highest level had a
    lower food consumption throughout the experiment than the controls
    with a significant decrease in body weight at 16 and 64 weeks. In
    females the relative weights of heart, liver and kidney were
    increased. No effects were found on histopathology and as regards
    haemoglobin levels (Allmark et al., 1957).

         Seventy-five rats were fed the colour at a level of 0.1% of the
    diet. No tumours were observed. Similar results were obtained with 10
    rats fed at 0.2% of the diet. Feeding extended for life span (DFG,

         Thirteen rats were given twice weekly s.c. injections of 0.5 ml
    of 1.0% of the colour for 365 days. Observation extended for 857 days.
    Five animals died during the experiment. No tumours were found (DFG,


         Several long-term studies specifically designed for
    carcinogenesis only have been performed in the rat, and another 80
    week study in the mouse is now available. Teratology has been examined
    in the mouse and a short-term study in a non-rodent species has been
    performed. There is little information on the metabolism of this


    Level causing no toxicological effect

         Mouse: 0.05% (= 500 ppm) in the diet equivalent to 25 mg/kg bw.

    Estimate of acceptable daily intake for man

         0-0.125 mg/kg bw*


         Required by June 1978

         Metabolic studies in several species preferably including man.
    Adequate long-term study in another species and reproduction studies.


    Allmark, M. G., Mannell, W. A. & Grice, H. C. (1957) J. Pharm.
         Pharmacol., 9, 622

    Bar, F. & Griepentrog, F. (1960) Med. u. Ernähr., 1, 99

    Deutsche Forschungsgemeinschaft, Farbstoff Kommission (1957)
         Mitteilung 6

    Gaunt, I. F. et al. (1967) Fd. Cosmet. Toxicol., 5, 187

    Gaunt, I. F. et al. (1969) Fd. Cosmet. Toxicol., 7, 443

    Larsson, K. S. (1974) Unpublished report submitted by National Food
         Administration, Stockholm, to WHO

    Lück, H. & Rickerl, E. (1960) Z. Lebensmitt.-Untersuch., 112, 157


    *    Temporary.

    Mason, P. L. et al. (1974) Fd. Cosmet. Toxicol. (In press)

    Ryan, A. J. & Wright, S. E. (1961) J. Pharm. Pharmacol., 13, 492

    Walker, R. (1968) Ph.D. Thesis University of Reading - cited by Gaunt,
         I. F. et al. (1969)

    See Also:
       Toxicological Abbreviations
       Ponceau 4R  (FAO Nutrition Meetings Report Series 46a)
       Ponceau 4R (WHO Food Additives Series 13)
       Ponceau 4R (WHO Food Additives Series 18)
       PONCEAU 4R (JECFA Evaluation)