Summaries of toxicological data: Toxicological tests on flavouring matters

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  • Fd Cosmet. Toxicol. Vol. 7, pp. 405--407. Pergamon Press 1969. Printed in Great Britain



    J. M. POSTERNAK and A. LINDER Institute of Physiology, School of Medicine and Laboratory of Mathematical Statistics, University of Geneva,

    Switzerland and

    C. A. VoDoz Firmenich & Cie, P.O.B. 39 Jonction, Geneva, Switzerland

    (Summary* of unpublished work commissioned by Firmenich & Cie, Geneva, Switzerland, and carried out in 1962-1967)


    Following the enactment of the US Food Additives Amendment in 1958, the Flavor and Extract Manufacturers' Association of the United States (FEMA) set up a Food Additives Committee which submitted to an Expert Panel of toxicologists and pharmacologists all known data on the uses, levels of use, metabolism and toxicity of flavouring substances. This Expert Panel considered whether each substance could be 'generally recognized as safe' (GRAS) under the intended conditions of use. The Panel's recommendations led to the clearance of many flavouring substances by the FDA under Section 121.1164 of the Code of Federal Regulations (Title 21, Chapter I). The data obtained from the work reported here have been submitted to the FEMA Expert Panel.

    With two exceptions, the dose levels used for these tests were calculated according to the FEMA method, described by Oser et al. (Fd Cosmet. Toxicol. 1965, 3, 563).

    A brief and partial summary of this work has already been published (Vodoz, Biblthca "Nutr. Dieta' 1967, 9, 87).

    Toxicity tests

    Test materials. The 42 commercially-available ttavouring matters that were tested in rats are listed in Table 1. Of these, 29 were at least 98 % pure and, with the exception of hexane- 3,4-dione (80-85 % pure), the purity of the remainder was 90 % or more.

    Experimental design and conduct. Groups of 10-16 male and equal numbers of female rats of the Charles River C.D. strain were housed in pairs of the same sex and fed for 90 days on a nutritionally-adequate basal diet (prepared by Veevoederfabrieken G.O.G., Doesburg, Netherlands or NAFAG, Gossau, Switzerland) alone or supplemented with one of the 42 flavouring matters (Table 1).

    * Prepared by BIBRA and published with the authors' permission. A detailed account of these investiga- tions will be published elsewhere.


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    The test materials were incorporated into the diet as 16.7 ~o emulsions in gum arabic, as 16-7 % adsorbates on microcrystalline cellulose, or as 10~o solutions in ethanol or peanut oil. The dietary levels of all but two of the flavourings were adjusted on two, three or four occasions during the 90-day study so that animals received on a mg/kg body weight basis a level in excess of 100 times the maximum estimated daily dietary intake of a 50-kg human (Table 1). In practice the 100-fold margin of intake was exceeded by 10-40 ~o.

    The body weight of individual rats and the food consumption of pairs of rats (as housed) were recorded weekly and the efficiency of food utilization was calculated. Haematological examination and blood urea determinations were carried out on 50yo of the animals at wk 7 and on all the animals at wk 13 of the test. At autopsy, the liver and kidneys were weighed and gross and histological examinations were carried out on a wide range of organs.

    Body weight and food intake. No significant differences were found between control and test rats in respect of growth, food intake and efficiency of food utilization throughout the 90-day study. Rats given compounds 108, 136 or 148 exhibited lower weight gains than the controls but when adjustments were made to initial body weights no significance could be attached to them.

    Haematology. There were no major differences in the haemoglobin concentration, erythrocyte count, haematocrit or total and differential leucocyte counts between control and test rats at wk 7 or 13. Although the observed values for some compounds (notably the differential leucocyte count at wk 7 for compound 114) fell slightly outside the control ranges, none of the deviations was regarded as being of pathological significance.

    Clinical chemistry. Blood-urea levels were raised in rats given compounds 102 (7 wk), 1 t l , 117, 129, 130 and 149 and were depressed in rats on compounds 95, 102 (13 wk), 104, 105, 147 and 151, but the differences were small when test values were compared with the composite rather than the individual control groups. The changes seen were therefore not considered to be toxicologically significant.

    Organ weights. Livers of male and female rats on compound 112 and of males on com- pound 111 were slightly enlarged and a similar effect was seen in the kidneys of males on compounds 107 and 108. However, these slight increases in relative weight became even less marked when test values were compared with composite control values and hence were deemed toxicologically insignificant.

    Organ pathology. Non-specific inflammatory changes were seen in the kidneys, fiver and, to a lesser extent, in the heart and lungs. However, the random distribution of these changes between control and test rats ruled out any causal connexion with the feeding of the test materials.


    None of the 42 flavouring matters fed for 90 days to rats at more than 100 times the equivalent of the human daily intake had any adverse effect on growth, food intake, haematological and clinical chemistry parameters, organ weights or organ pathology.

    Acknowledgements--The anatomo- and histopathological investigations were performed by Dr. Th. Rabino- wicz, privat-docent at the University of Lausanne, whom we thank for his highly competent collaboration. We are grateful to Miss E. Schmidt, head technician of our laboratory, for.her dedication to this work and for her skilful assistance.


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