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    INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY

    WORLD HEALTH ORGANIZATION



    TOXICOLOGICAL EVALUATION OF SOME
    FOOD COLOURS, EMULSIFIERS, STABILIZERS,
    ANTI-CAKING AGENTS AND CERTAIN
    OTHER SUBSTANCES



    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.


    PATENT BLUE V

    Biological Data

    Biochemical aspects

    After intravenous injection into the rat of 0.5 ml of an aqueous
    solution containing 5 g per 100 ml the colour is excreted in the urine
    which is coloured blue during the 12 hours following the injection.
    Findings are similar in the case of man when the colour is injected
    subcutaneously for the detection of peripheral lymphatic trunks and
    nodes for the purpose of lymphography. (Truhaut, 1962).

    Acute toxicity

                                                                     

    Animal    Route          LD50                 Reference
                        per kg body-weight 
                                                                     

    Mouse    oral           > 3.0 g               Truhaut, 1962

    Mouse    i.v.             1.2 g                    "

    Rat      oral           > 5.0 g                    "

    Rat      i.v.             5.0 g                    "
                                                                     

    Special studies

    The dye (as the Ca salt) had slight surface activity. The Na salt had
    considerable surface activity. Protein binding and lipophilic activity
    were negligible for both salts. The authors conclude that the
    difference in surface activity explains the difference in the local
    reactions to the two salts (Cangolli et al., 1967).

    Short-term studies

    Rat. Subcutaneous injections of 1 ml of 0.8 per cent. aqueous
    solution twice weekly for five weeks produced only slight tissue
    reactions (Grasso & Goldberg, 1966).

    Cat. Three cats were given daily a five per cent. aqueous solution
    of the colour orally doses of 0.25, 0.50 and 0.75 g. No abnormalities
    were found (Truhaut, 1962).

    Long-term studies

    Rat. Sixty rats, half males and half females, were given in their
    diet 10 000 ppm of the colour for their life-span.  The used colour
    was pure (no impurities were found by paperchromatography studies).
    The average life-span of treated animals was 24 months and the last
    animal died at an age of 37 months, two weeks. The average intake of
    the colour was approximately 8O g. Forty animals were used as
    controls.The average life-span of these animals was 22 months, two
    weeks. For the second experiment 30 rats, half males and half females,
    were given the same diet with 10 000 ppm and also for 15-19 months
    once a week a subcutaneous injection of 1 ml of a one per cent.
    aqueous solution. The average life-span of the animals was 18 months.
    The last animal died at an age of 30 months. Twenty-six rats were used
    as controls and were given 1 ml of distilled water subcutaneously for
    the same period as the treated animals. The average life-span of these
    animals was 19 months.

    In both experiments it was found that growth, blood composition,
    reproduction and the three generations of animals which were bred were
    not influenced. Also, no abnormal histopathological findings or
    sarcomas at the site of injection were observed (Truhaut, 1962).

    Comments

    Information on the metabolism of the colour is lacking. Long-term and
    reproduction studies in the rat did not reveal any significant
    toxicological effects. However, the corresponding sodium salt which is
    not used as food colour showed evidence of considerable surface
    activity on subcutaneous injection and for this reason a long-term
    study in a second species is required.

    EVALUATION

    Level causing no toxicological effects in the rat

    One per cent. (= 10 000 ppm) in the diet equivalent to 500 mg/kg body
    weight/day.

    Estimate of daily acceptable intake for man

                               mg/kg body weight

    Temporary acceptance             0 - 1

    Further work required by June 1974

    Metabolic studies in several species, preferably including man, two
    year studies in non-rodent mammalian species and long-term studies in
    a second species.

    REFERENCES

    Gangolli, S. D., Grasso, P. & Go1dberg, L. (1967) Fd. Cosmet.
    Toxicol., 5, 601

    Grasso, P. & Goldberg, L. (1966) Fd. Cosmet. Toxicol., 4, 269

    Truhaut, R. (1962) Aliment. et Vie, 50, 77
    


    See Also:
       Toxicological Abbreviations
       Patent blue V (WHO Food Additives Series 6)
       PATENT BLUE V (JECFA Evaluation)