Ethyl vanillin was evaluated at the eleventh meeting of the
    Committee (Annex 1, ref 14) when an Acceptable Daily Intake of 0-10
    mg/kg bw was allocated to this compound.  The eleventh meeting noted
    that few metabolic studies had been carried out on this compound and
    considered that further metabolic studies were desirable.

         Ethyl vanillin was placed on the agenda of the present meeting on
    the basis of partial application of the method for setting priorities
    for the safety review of food flavouring ingredients (Annex 1,
    reference 83).

         Since the last review, additional data have become available and
    are summarized and discussed in the following monograph.  The
    previously published monograph has been expanded and is incorporated
    into this monograph.


    2.1  Biochemical aspects

    2.1.1  Metabolism and Pharmacokinetics

         This compound is probably metabolized to glucuroethyl vanillin
    and ethyl vanillic acid, of which some is conjugated with glucuronic
    and sulfuric acids (Williams, 1959).

    2.2  Toxicological studies

    2.2.1  Acute toxicity


    Species        Route         LD50              Reference
                              (mg/kg bw)

    Mouse          i.p.           750       Caujolle & Meynier, 1954a

    Rat            oral          1590       Sporn, 1960
                   oral         >2000       Jenner  et al., 1964
                   s.c.          1800       Deichmann & Kitzmuller, 1940

    Guinea pig     i.p.          1140       Caujolle & Meynier, 1954b

    Dog            i.v.           760       Caujolle  et al., 1953

    * Maximum non-lethal dose, 450 mg/kg bw; LD50 = 950 mg/kg bw.
         A dose of 150 mg/kg bw caused no adverse effects and at 2500
    mg/kg bw only a transient increase in respiration rate was observed
    when groups of 6 rabbits were given ethyl vanillin by oral gavage. 
    The minimum oral lethal dose was reported to be 3000 mg/kg bw
    (Deichmann & Kitzmuller, 1940).

    2.2.2  Short-term studies  Rats

         Doses of 300 mg/kg bw were administered to rats by gavage twice
    weekly for 14 weeks without any adverse effects.  In another
    experiment, groups of 16 rats were fed 20 mg/kg bw daily for 18 weeks
    without adverse effect but 64 mg/kg bw daily for 10 weeks reduced
    growth rate and caused myocardial, renal, hepatic, lung, spleen and
    stomach injuries (Deichmann & Kitzmuller, 1940).

         Groups of male rats were fed 0, 2 or 5% ethyl vanillin in the
    diet for one year without any adverse effects (Hagan  et al., 1967).

         Sixteen rats were given 30 mg ethyl vanillin weekly for 7 weeks
    without adverse effect on growth, food intake or protein utilization
    (Sporn, 1960).  Rabbits

         Single rabbits were given ethyl vanillin orally in 10% aqueous
    glycerine at doses of 15 mg/kg bw/day for 13 of 15 days, or 26 of 31
    days, 32 mg/kg bw/day for 15 of 17 days, 41 mg/kg bw/day for 26 of 31
    days or 49 mg/kg bw/day for 43 of 49 days.  At the highest dose level,
    anaemia, diarrhoea and lack of weight gain were observed but no toxic
    signs were reported at any of the lower levels (Deichmann &
    Kitzmuller, 1940).

    2.2.3  Long-term/carcinogenicity studies  Mice

         The maximum tolerated dose for ethyl vanillin in mice when
    administered i.p. 3 times per week for 2 weeks and tolerated through
    a subsequent 24 week observation period was reported to be 75 mg/kg
    bw.  Administration of ethyl vanillin i.p. at doses of 15 or 75 mg/kg
    bw three times a week for eight weeks resulted in respective
    mortalities of 8/20 and 10/20 animals.  Control animals receiving i.p.
    injections of the vehicle, tricaprylin, had survival rates of 77/80
    (males) and 77/80 (females).  In the control group, 28% of males and
    23% of females developed lung tumours whereas in the treated groups
    only one animal, in the higher dose group, exhibited a single lung
    nodule.  It was concluded that ethyl vanillin did not potentiate the
    pulmonary tumour responses in strain A mice (Stoner  et al., 1973).  Rats

         Groups of 12 male and 12 female rats were fed diets containing 0,
    0.5, 1 and 2% ethyl vanillin for 2 years or 2% and 5% for one year
    without any adverse effects on growth, organ weights of major organs,
    hematology and histology of major tissues (Hagan  et al., 1967).

    2.2.4  Reproduction studies

         No data available.

    2.2.5  Special studies on mutagenicity

         Ethyl vanillin was negative in a mouse micronucleus assay when
    given to groups of four mice as two i.p. doses of up to 1 g/kg bw at
    an interval of 24 hours (Wild  et al., 1983).

         Ethyl vanillin did not induce mutations in the Ames  Salmonella/
    microsome assay against  Salmonella typhimurium strains TA92, TA1535,
    TA100, TA1537, TA94 and TA98, with or without metabolic activation
    using rat or hamster liver S9 fractions (Wild  et al., 1983;
    Mortelmans  et al., 1986).

         In  in vitro assays using Chinese hamster ovary cells, ethyl
    vanillin did not induce chromosomal aberrations; the compound did
    cause an increase in polyploid cells but the significance of this is
    unclear and similar polyploidy was induced by riboflavin (Ishidate
     et al., 1984).

         Ethyl vanillin did not induce sister chromatid exchanges in
    cultured Chinese hamster ovary cells  in vitro but was reported to
    enhance the ability of mitomycin C to cause sister chromatid exchanges
     (Sasaki et al., 1987).

         No heritable mutations were detected when ethyl vanillin was used
    in the BASC on  Drosophila melanogaster (Wild  et al., 1983).

         In a study on the anti-mutagenic potential of flavourings, ethyl
    vanillin was reported to show marked anti-mutagenic activity against
    mutagenicity induced by 4-nitroquinoline-1-oxide, furylfuramide,
    captan or methylglyoxal in  Escherichia coli WP2s but was ineffective
    against mutations induced by Trp-P-2 or IQ in  Salmonella typhimurium
    TA98.  It was assumed that the anti-mutagenic activity was due to
    enhancement of an error-free recombinational repair system (Ohta
     et al., 1986).

    2.3  Observations in man

         In a 24-hour closed patch test in 25 subjects, ethyl vanillin
    tested at 2% in petrolatum produced a mild irritation.  No
    sensitization reactions were evoked when ethyl vanillin was used at 2%
    in petrolatum in a maximization test on 25 volunteers (Kligman, 1970).


         The Committee noted that none of the previously evaluated long-
    term or carcinogenicity studies meet modern standards in that fewer
    animals per group had been used than would be the present norm. 
    Accordingly the Committee reduced the previous ADI and made it


         Level causing no toxicological effect
         Rat: 5000 ppm, equivalent to 250 mg/kg bw/day.

         Estimate of temporary acceptable daily intake
         0-5 mg/kg bw.

         Further work required by 1992
         1.   An adequate short-term study in rats, and
         2.   metabolic studies in rats.


    CAUJOLLE, F. & MEYNIER, D. (1954a). Toxicity of vanillin, o-vanillin
    and ethyl vanillin and of the corresponding meta aldehydes.
     Ann.Pharm.Fr., 12, 42-49.

    CAUJOLLE, F. & MEYNIER, D. (1954b). The comparative toxicities of
    orthovanillin and ethyl orthovanillin.  Compt.Rend., 238, 2576-2578.

    CAUJOLLE, F., MEYNIER, D. & MOSCARELLA, C. (1953). The comparative
    toxicity of ethyl vanillin and 4-hydroxy-5-ethoxyisophthalic
    dialdehyde.  Compt.Rend., 237,765-766.

    DEICHMANN, W. & KITZMULLER, K.V. (1940). Toxicity of vanillin and
    ethyl vanillin for rabbits and rats.  J.Am.Pharm.Assoc., 29, 425-428.

    TAYLOR, J.M., LONG, E.L., NELSON, A.A. & BROUWER, J.B. (1967). Food
    flavourings and compounds of related structure. II. Subacute and
    chronic toxicity.  Fd Cosmet.Toxicol., 5, 141-157.

    SAWADA, M., & MATSUOKA. A. (1984). Primary mutagenicity screening of
    food additives currently used in Japan.  Fd Chem.Toxicol., 22,

    (1964). Food flavourings and compounds of related structure. I. Acute
    oral toxicity.  Fd Cosmet.Toxicol., 2, 327-343.

    KLIGMAN, A.M. (1970). Unpublished report to RIFM cited in Opdyke,
    D.L.J. (1975). Monographs on fragrance raw materials.
     Fd Cosmet.Toxicol., 13, 103-104.

    MORTELMANS, K. et al., (1986).  Environ.Mutagen., 8, Suppl.7, 1.

    OHTA, T., WATANABE, M., WATANABE, K., SHIRASU, Y. & KADA, T. (1986).
    Inhibitory effects of flavourings on mutagenesis induced by chemicals
    in bacteria.  Fd Chem.Toxicol., 24, 51-54.

    SASAKI, Y.F., IMANISHI, H., OHTA, T. & SHIRASU, Y. (1987). Effects of
    antimutagenic flavourings on SCEs induced by chemical mutagens in
    cultured Chinese hamster cells.  Mutation Res., 189, 313-318.

    SPORN, A. (1960).  Igiena (Bucharest), 9, 365.

    WEISBURGER, E.K. & GORI, G.B. (1973). Test for carcinogenicity of food
    additives and chemotherapeutic agents by the pulmonary tumour response
    in strain A mice.  Cancer Res., 33, 3069-3085.

    WILD, D., KING, M-T., GOCKE, E. & ECKHARDT , K. (1983). Study of
    artificial flavouring substances for mutagenicity in the
    salmonella/microsome, BASC and micronucleus tests.  Fd Chem.Toxic.,
    21, 707-719.

    WILLIAMS, R.T. (1959).  Detoxication Mechanisms, 2nd Edition, Chapman
    & Hall, London.

    See Also:
       Toxicological Abbreviations
       Ethyl vanillin (FAO Nutrition Meetings Report Series 44a)
       Ethyl vanillin (WHO Food Additives Series 35)
       ETHYL VANILLIN (JECFA Evaluation)