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    INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY

    WORLD HEALTH ORGANIZATION



    TOXICOLOGICAL EVALUATION OF SOME
    FOOD COLOURS, ENZYMES, FLAVOUR
    ENHANCERS, THICKENING AGENTS, AND
    CERTAIN FOOD ADDITIVES



    WHO FOOD ADDITIVES SERIES 6







    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.

    FUMARIC ACID

    Explanation

         This compound has been evaluated for acceptable daily intake by
    the Joint FAO/WHO Expert Committee on Food Additives (see Annex 1,
    Ref. No. 13) in 1965.

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

    BIOLOGICAL DATA

    BIOCHEMICAL ASPECTS

         Fumaric acid is a normal constituent of tissues as an
    intermediate in the tricarboxylic acid cycle. Distribution of fumaric
    acid in rat tissue has been studied by partition chromatography and it
    was found that blood contained 3 mg/l, brain tissue 150 mg/kg, kidney
    tissue 95 mg/kg, liver 78 mg/kg and muscle 23 mg/kg (Marshall et al.,
    1949).

         The laxative effect of fumarates was studied in mice by
    determining the dose which shortened the average time before soft
    faeces appeared after oral administration. 433 mg magnesium fumarate
    or 432 mg disodium fumarate shortened the average time by 40%. If the
    appearance of charcoal marker was taken as a guide, 15% shortening of
    appearance time was effected by 843.6 mg magnesium fumarate or 640 mg
    disodium fumarate (Locke et al., 1942). Twenty-six constipated
    patients suffering from a variety of chronic diseases not involving
    the gastrointestinal tract were given oral doses of 5-30 g sodium
    fumarate, a satisfactory bowel motion resulting in 18 patients. There
    was much variability of response to a given dose between patients and
    in the same individual. Doses above 15 g caused unpleasant side
    effects. No abnormalities were noted in the urine or serum non-protein
    nitrogen level (Bodansky et al., 1942).

    TOXICOLOGICAL STUDIES

    Acute toxicity
                                                                        

                                     LD50
    Animal   Route                   (mg/kg bw)       References
                                                                        

    Rat      Oral (sodium fumarate)  approx. 8 000    Levey et al., 1946

    Rabbit   Oral (disodium          approx. 3 600    Locke et al., 1942
             fumarate)                       4 800    Weiss et al., 1923
                                                                        

    Short-term studies

    Guinea-pig

         Eight animals were maintained on a diet containing 0%, 1% and 
    10% fumaric acid for one year without any adverse effect on growth. 
    The second generation from four mated animals was treated similarly 
    without any adverse effect noted regarding growth, fertility or 
    lactation (Levey et al., 1946).

    Rabbit

         Each of five rabbits received i.v. injections of 50-500 mg/kg
    sodium fumarate every second or third day for 10-32 days without any
    injurious effect on blood levels of non-protein nitrogen or
    creatinine, phensulfolphthalein excretion, or kidney and liver
    histology (Bodansky et al., 1942). Six rabbits received twice weekly
    i.p. injections of 60 mg/kg bw of sodium fumarate over 17-29 weeks.
    Swelling and congestion of the thyroids and atrophy of testes, with
    low hyaluronidase content, were found (Arai & Suchiro, 1953). A
    further nine male rabbits received 60 mg/kg bw sodium fumarate every
    second day by i.p. injection for 150 days. By the end of the test
    period, gonadotropic activity of the serum, as well as oestrogenic
    activity was detected. There was progressive testicular atrophy in all
    animals, resulting in disappearance of seminiferous epithelium and
    survival of Sertoli cells only. Chromophobe cells were increased in
    the pituitary (Arai et al., 1955).

         Fourteen rabbits were fed 320-2080 mg/kg bw of disodium fumarate
    daily for 28 days without any deaths. A further six rabbits received
    2880-3680 mg/kg bw for 17 days with three deaths. Two rabbits were fed
    a daily diet containing 640 mg/kg bw for 36 days without consistent
    adverse effect on body weight, haematology, non-protein nitrogen or
    creatinine levels, or histopathological findings (Locke et al., 1942).
    In another experiment, four groups of 15 rabbits were fed diets

    containing 0 or 6.9% sodium fumarate (equivalent to 5% fumaric acid)
    for 150 days. There were no significant differences from controls in
    body weight gain, food consumption, mortality rate, blood counts,
    blood sugar, non-protein nitrogen level and urine. Organ weights were
    not significantly different between the groups and histologic
    examination showed no adverse findings attributable to the diet. In
    particular, spermatogenesis and testicular structure were unaffected
    (Packman et al., 1963).

    Dog

         Fumaric acid was fed to four groups of six young dogs at 0, 1, 3
    and 5% of the diet for two years without adverse effect on body weight
    gain, development, haematology, blood sugar and urea levels,
    haemoglobin and urine. Organ weights and gross and histopathological
    examination of all principal organs and tissues revealed no effects
    attributable to the treatment (Harrisson & Abbott, 1962).

    Long-term studies

    Rat

         Eight groups of 14 weanling rats were kept on diets containing 0,
    0.1 and 1.0% fumaric acid and 1.38% sodium fumarate for one year (half
    the groups) or two years. No adverse effect was noted on rate of
    weight gain, haemoglobin, blood picture, calcium balance as shown by
    bone histology, or on the histology of liver, kidney, spleen and
    stomach (Levey et al., 1946).

         In another experiment five groups of 12 male and 12 female rats
    were fed diets containing 0, 0.1, 0.5, 0.8 and 1.2% of fumaric acid
    for two years without toxic effects on growth or food consumption. A
    further four groups of 12 male rats were kept for two years on diets
    containing 0, 0.5, 1.0 and 1.5% fumaric acid. Only at the 1.5% level
    was there a very slight increase in mortality rate and some testicular
    atrophy. Gross and microscopic examination of major organs revealed no
    abnormalities and tumour incidence was not significantly different
    between the groups (Fitzhugh & Nelson, 1947).

    OBSERVATIONS IN MAN

         Seventy-five chronically disabled subjects ranging in age from
    29-91 years received 500 mg fumaric acid daily for one year without
    any toxic manifestations in haemoglobin level, RBC and WBC, non-
    protein nitrogen level, creatinine level, bromosulfonphthalein
    excretion and phenolsulfonphthalein excretion (Levey et al., 1946).

    Comments:

         Fumaric acid is a normal component of intermediary metabolism.
    The testicular atrophy in rabbits reported after intraperitoneal
    administration of high doses was not seen after oral administration of
    doses as high as 6-9% in the diet in rabbits and other species.

    EVALUATION

    Level causing no toxicological effect

         Rat: 1.2% (=12 000 ppm) in the diet equivalent to 600 mg/kg bw.

    Estimate of acceptable daily intake for man

         0-6 mg/kg bw

    REFERENCES

    Arai, T. & Suchiro, S. (1953) Wakayama med. Rep., 1, 35

    Arai, T., Suchiro, S. & Okamoto, T. (1955) Wakayama med. Rep., 2, 115

    Bodansky, O., Gold, H. & Zahm, W. (1942) J. Amer. pharm. Ass., sci.
         Ed., 31, 1

    Fitzhugh, O. G. & Nelson, A. A. (1947) J. Amer. pharm. Ass., sci. Ed.,
         36, 217

    Hall, R. L. (1960) Food Techn., 14, 488

    Harrison, J. W. E. & Abbott, D. D. (1962) Unpublished report of LaWall
         and Harrisson Research Laboratories, submitted to WHO

    Innes, J. M. (1936) Biochem. J., 30, 2040

    Levey, S. et al. (1946) J. Amer. pharm. Ass., sci. Ed., 35, 298

    Locke, A. et al. (1942) J. Amer. pharm. Ass., 31, 12

    Marshall, L. M., Orten, J. M. & Smith, A. H. (1949) J. biol. Chem.,
         179, 1127

    Packman, E. W., Abbott, D. D. & Harrisson, J. W. E. (1963) Toxicol.
         appl. Pharmacol., 5, 163

    Weiss, J. M., Downs, C. R. & Corson, H. P. (1923) Ind. Eng. Chem., 15,
         628


    See Also:
       Toxicological Abbreviations
       Fumaric acid (ICSC)
       Fumaric acid (FAO Nutrition Meetings Report Series 40abc)
       FUMARIC ACID (JECFA Evaluation)