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



         This food colour has been evaluated for acceptable daily intake
    by the Joint FAO/WHO Expert Committee on Food Additives (see Annex I,
    Refs 8 and 10) in 1964.




         Rats received orally 20 or 200 mg benzyl violet dissolved in
    10 ml of water. Urine and faeces were collected during 36 hours. With
    the faeces respectively 90 and 98% of the administered doses was
    excreted. In the urine no colouring could be found (Hess and Fitzhugh,

         Less than 5% of the administered benzyl violet was found in the
    bile of dogs after an oral administration of 200 mg per animal (Hess
    and Fitzhugh, 1955). Other aspects are discussed by Minegishi and
    Yamaha (1977).

         Despite low degree of absorption, unlike some colourings, violet
    6B does not have a cathartic effect (Radomski and Deichmann, 1956).


         Nature of interaction between serum proteins and violet 6B was
    investigated by electrophoresis.

         Electropherogram on cellulose acetate showed migration to the
    anode in a single band. The complex between serum proteins and violet
    6B were not separated by electrophoresis on cellulose acetate or
    polyacrylamide gel, but could be separated by filtration on Sephadex
    gel (Gangolli et al., 1972).


    Acute toxicity


    Species    Route      LD50 mg/kg bw    References

    Mouse      oral          4 000         Tullar, 1947
    Mouse      i.p.            432         Tullar, 1947
    Mouse      i.v.            141         Tullar, 1947
    Rat        oral          2 000         Lu et al., 1964
    Rabbit     i.p.            500         Tullar, 1947
    Rabbit     i.v.            150         Tullar, 1947

    Short-term studies

         Rat Groups of 15 male and 15 female rats (strain Wistar) were
    given diets containing 0 (control), 0.5, 1.0 or 3% violet 6B (purity
    min. 85%) for 13 weeks. There was marked excretion of the colour in
    the faeces indicating a low level of absorption. No statistically
    significant effects were noted in body weight gain, haematologieal
    values, serum or urine analysis or renal function tests. There were no
    adverse histopathological findings attributable to the feeding of the
    colouring. The weights of heart, liver and coecum were lower in the
    rats given 3% of the dye than the controls and the relative kidney
    weights were higher. The no-effect level in this study was 1% (Gaunt
    et al., 1974).


         Groups of two male and two female beagles were fed at levels of
    0.5, 1.0 and 2.0% of the colour in the diet for two years. Of each
    group one dog died during the experiment. No pathological changes to
    account for death could be found. Dose-related changes were a tan
    non-icteric tinge of the subcutaneous tissue and (microscopically
    confirmed) hyperkeratosis of the ears (USFDA, 1964) (no details

    Long-term studies


         Groups of 48 male and 50 female mice (strain ASH-CS1) were fed
    diets containing 0, 70, 700 or 3500 ppm violet 6B (purity min. 85%)
    for 80 weeks. The faeces of all treated mice were stained but the
    urine appeared normal suggesting little absorption from the GI tract.

    There were no effects on mortality, rate of body weight gain or
    haematology at 12, 28, 52 and 80 weeks. No adverse findings including
    tumours were seen in treated compared with control animals. Except in
    the females with 700 and 3500 ppm a slight increase in lymphosarcoma
    in the thymus was found. Whether this finding is relating with the
    administration of the dye is not known. It was concluded that feeding
    up to 3500 ppm did not exert carcinogenic effect (Grasso et al., 1971,


         Eighteen young rats were given 20 mg of the colour (2% aqueous
    solution) by subcutaneous injections once a week for two years; 14
    fibrosarcomas were induced (Nelson and Davidow, 1957).

         Four groups of 15 male and 15 female rats (strain Wistar)
    received during 75 weeks respectively 0 (control), 0.03, 0.3 and 3.0%
    of benzyl violet (purity not given) in their diet. No depression of
    growth, food consumption or food efficiency was found. Haematology did
    not show abnormalities. Seven males and four females of the 0.03%, one
    male and three females of 0.3% and four males and seven females of 3%
    level died (in control group six males and one female), 3/7 females on
    3% level that died during the experiment showed tumours (none in the
    controls). Five male and five female rats per group were examined
    histologically after 75 weeks. 1/5 male and 4/5 female rats of 3%
    level showed malignant tumours (one male at controls). The incidence
    of the skin tumours in rats of the 3% level was 20% (possibly from
    contact of the fur and skin of the rats with the food) (Mannell et
    al., 1962).

         Groups of 15 litter-mated male rats were fed the colour at 0.1%,
    0.5% and 1.0% in the diet for two years. The only pathological effect
    noted was slight dilation of the gastric glands at the 1.0% level
    (FDA, 1964) (no details available).

         Groups of 20 male and 20 female rats were kept in such a way that
    the skin was in regular contact with the colour for 88-100 weeks.
    Similar groups served as control. The experimental animals showed a
    small number of ulcerative dermatitis of the scrotum, possibly as a
    result of a chronic irritation caused by contact of the scrotal skin
    with the floor of the cage and the colour. No malignant tumours of the
    epidermis were found, but there was an increase in benign mammary
    tumours (Mannell et al., 1964).

         Twenty males and 20 females Sprague-Dawley rats were fed 5% of
    acid violet 6B (purity 89.1%) for one year, 10 control males and 10
    control females received basic diet only. Mortality was greater in
    treated animals, five mammary carcinomas and four induced earduct
    carcinomas were found in 18 female animals, one mammary fibroadenoma

    occurred in female controls. No metastases were found. At one year
    only one test male and one test female and five control females
    survived. No tumours were found in the males (Uematsu and Miyaji,

         Two groups of 35 female (Sprague-Dawley) rats received during 12
    months respectively normal diet and a diet containing first 1%,
    thereafter 3%, each for one week respectively and then maintained on
    5% of benzyl violet 4B (purity 89.4%). The mortality in the test group
    was increased significantly. Only two rats survived 12 months, in the
    control group all rats survived. Growth of the rats of the test group
    was reduced significantly, though food consumption was normal. In
    22/35 rats in the experimental group, tumours developed externally.
    Eleven rats with mammary gland carcinoma, four rats with squamous cell
    carcinoma of the skin and seven rats with both types of tumours were
    seen. Hyperplasia of sebaceous glands was observed in the skin at the
    site of development of both types of tumours. The onset of tumour
    development was earlier for the mammary gland carcinoma. The majority
    of the squamous cell carcinomas developed in the earduct, but they
    also appeared in the buccal and axillary regions. In the control group
    no external tumours were noted. Macroscopy did not reveal nodules,
    suggestive of metastasis of external tumours to internal organs (Ikeda
    et al., 1974).


    Gangolli, S. D., Grasso, P., Golberg, L. and Hooson, J. (1972) Protein
    binding by food colourings in relation to the production of
    subcutaneous sarcoma, Food and Cosm. Tox., 10, 449-462

    Gaunt, I. F., Hardy, J., Kiss, I. S. and Gangolli, S. D. (1974)
    Shortterm toxicity of violet 6B (FD and C Violet no. 1) in the rat,
    Food and Cosm. Tox., 12, 11-19

    Grasso, P., Hardy, J., Gaunt, I. F., Mason, P. L. and Lloyd, A. G.
    (1974) Longterm toxicity of violet 6B (FD and C Violet no. 1) in mice,
    Food and Cosm. Toxicol., 12, 21-31

    Grasso, P. et al. (1971) Food and Cosm. Toxicol., 9, 463-478

    Hess, S. M. and Fitzhugh, O. G. (1955) Absorption and excretion of
    certain triphenylmethane colours in rats and dogs, J. Phamac.
    exp. Ther., 114, 38-42

    Ikeda, Y., Horiuchi, S., Imoto, A., Kodama, Y., Aida, Y. and
    Kobayashi, K. (1974) Induction of mammary gland and skin tumours in
    female rats by the feeding of henzylviolet 4B, Toxicology, 2,

    Lu, F. C. and Lavallee, A. (1964) Canad. pharm. J., 97, 30-

    Mannell, W. A., Grice, H. C. and Allmark, M. G. (1962) Chronic
    toxicity studies on food colours. V. Observations on the toxicity of
    brilliant blue FCF, guinea green B and benzylviolet 4B in rats,
    J. Pharm. Pharmacol., 14, 378-384

    Mannell, W. A., Grice, H. C. and Dupuis, J. (1964) The effect on rats
    of longterm exposure to guinea green B and benzylviolet 4B, Food and
    Cosm. Tox., 2(3), 345-347

    Minegishi, K. and Yamaha, T. (1977) Toxicology, 7, 367

    Nelson, A. A. and Davidow, B. (1957) Fedn. Proc. Fedn. Am. Socs.
    Exp. Biol., 16, 367

    Radomski, J. L. and Deichmann, W. B. (1956) J. Pharmac. Exptl.
    Ther., 118, 322

    Tullar, P. E. (1947) Report dated 1.10.1947 from George Washington
    University (unpublished report)

    Uematsu, K. and Miyaji, T. (1973) Induction of tumours in rats by oral
    administration of technical acid violet 6B, J. Nat. Canc. Inst.,
    51, 1337-1338

    United States Food and Drug Administration (1964) Unpublished report
    submitted to WHO

    See Also:
       Toxicological Abbreviations
       BENZYL VIOLET 4B (JECFA Evaluation)
       Benzyl Violet 4B  (IARC Summary & Evaluation, Volume 16, 1978)