FAO, PL:CP/15
    WHO/Food Add./67.32


    The content of this document is the result of the deliberations of the
    Joint Meeting of the FAO Working Party and the WHO Expert Committee on
    Pesticide Residues, which met in Geneva, 14-21 November 1966.1

    1 Report of a Joint Meeting of the FAO Working Party and the WHO
    Expert Committee on Pesticide Residues, FAO Agricultural Studies, in
    press; Wld Hlth Org. techn. Rep. Ser., 1967, in press




    EDB, ethylene bromide

    Chemical name





    Biochemical aspects

    The fate of ethylene dibromide absorbed from the mammalian alimentary
    tract has apparently not been determined. After uptake by the
    pulmonary route, however, some of the compound is excreted through the
    lungs and some of it is changed in the body with the formation of
    organic bromides (von Oettingen, 1955).

    Acute toxicity

    Animal            Route              LD50                  Reference
                                   mg/kg body-weight

    Mouse, female     Oral                420              Rowe et al., 1952

    Rat, male           "                 146                      "

    Rat, female         "                 117                      "

    Chick               "                 79                       "

    Guinea-pig          "                 110                      "

    Rabbit, female      "                 55                       "
    A 43-year-old woman who took a single dose of ethylene dibromide
    amounting to 4.5 ml died 54 hours later after symptoms of vomiting,
    diarrhoea and anuria. At autopsy there was massive centro-lobular
    necrosis of the liver and proximal tubular damage in the kidney
    (Olmstead, 1960).

    Short-term studies

    A total of 19 rats and guinea-pigs was fed ethylene dibromide in oil
    or 50 per cent alcohol for about 4 months and there were no abnormal
    signs in those receiving about 40-50 mg/kg/day, but no pathological
    tests or autopsy examinations appear to have been undertaken (Aman et
    al., 1946).

    Fumigants in 50 per cent aqueous ethanol solution were administered
    directly into the crops of adult hens. Individually caged birds were
    given respectively 0.5, 1, 2, 4 and 20 mg ethylene dibromide/hen/day
    for 8 weeks. An additional group was given 8 mg/hen/day together with
    ethylene dichloride and carbon tetrachloride at levels which had no
    effect when given alone. At 8 mg/day (with other fumigants) and 20
    mg/day both egg production and egg weight were decreased, whilst at 2
    and 4 mg/day only egg weight was depressed. At 0.5 and 1.0 mg/day
    there were no differences from the controls. Haugh units of quality
    and shell thickness were also determined in this study, and the
    fumigant had no effect on these parameters (Fuller & Morris, 1962).

    In another experiment, groups of 24 pullets were treated for 12 weeks,
    rested for 8 weeks, and treated again for 12 weeks. Doses were 0, 0.5,
    1, 2, 4 and 8 mg/bird/day. Egg production was depressed at 8 mg/day
    only. Egg weight was reported to be significantly decreased at all
    levels. However, the decrease at 0.5 and 1 mg/bird/day was very small.
    At necropsy, the birds receiving 8 mg/day and showing lowered
    productivity were found to have an excessive number of incompletely
    developed ovarian follicles, but no abnormalities in liver, brain or
    kidney. Effects on egg production and egg weight were comparable among
    two groups of hens treated at 7 and 18 months of age respectively
    (Fuller & Morris, 1962).

    Two-week-old chicks, in groups of 24, were fed for two weeks on a
    ration to which ethylene dibromide in ethanol and olive oil had been
    added at levels of 20, 40 and 80 ppm. Estimates made from chemical
    analyses suggested that these amounts would be reduced by evaporation
    and by conversion to inorganic bromide to somewhat less than 10, 20
    and 40 ppm by the time the ration was consumed. At 10 and 20 ppm no
    change was observed in the chicks over the two weeks, but at 40 ppm
    there was retardation of growth rate, even when a correction was made
    for food intake (Morris & Fuller, 1963).

    Six-month-old pullets in groups of 10 were given ethylene dibromide
    either directly by injection into the crop, or indirectly in the diet
    after fumigation, daily intakes being 0.5, 1, 2, 4 and 8 mg per day.

    At 8 mg per day by both routes of administration, egg production was
    depressed, while egg weight was reduced at all levels down to 0.5 mg
    per bird, whether by crop or diet (Fuller & Morris, 1963).

    Four bull calves were given a dose schedule averaging 2 mg/kg
    body-weight daily of ethylene dibromide from the age of four days.
    After 14-16 months, growth, health and libido of the treated bulls
    were comparable to the controls, but sperm density was low and
    motility poor. These changes were reversible (Amir & Volcani, 1965).

    Short-term studies with fumigated foodstuff

    When chickens were fed for five days, pigs for 12 days and heifers for
    one week on grain previously fumigated with a gaseous mixture
    containing 7.2 per cent ethylene dibromide none of them showed any
    deleterious effects (Rowe et al., 1954).

    Twenty-five adult hens, in groups of five, were given a ration made up
    of 50 per cent sorghum. In those groups receiving grain containing
    sorbed ethylene dibromide in amounts ranging from 50 to 320 ppm, there
    was a decrease in egg weight proportional to the level of the
    fumigant. Moreover, in those receiving the highest amount, egg laying
    ceased completely in six-and-a-half weeks, and in those receiving 200
    ppm it ceased within eight weeks. Even as little as 50 ppm had a
    depressive effect on egg size within three weeks. Further, those hens
    in which laying was arrested failed to resume when returned to a
    normal ration (Bondi et al., 1955).

    In another experiment with six-month-old hens in groups of 24 the
    sorghum, making up 50 per cent of the grain fed, contained 10-15 ppm
    ethylene dibromide and 20 ppm residual bromide, 20-30 ppm ethylene
    dibromide and 50 ppm residual bromide and no free ethylene dibromide
    but 120 ppm residual bromide. The fourth group was a control. Over 16
    weeks the group receiving only the residual bromide behaved the same
    as the controls, but those receiving ethylene dibromide showed a
    decrease in egg size proportional to dose. But when these birds were
    returned to a normal diet the egg size was regained (Bondi et al.,

    In an experiment carried out with two groups each of five hens, all of
    which were laying small eggs (40 per cent below normal) by being fed
    grain containing ethylene dibromide, the administration to one group
    of follicle stimulating hormone intravenously led to a partial
    recovery in egg size. Moreover, there was no reaction between ethylene
    dibromide and follicle stimulating hormone in sorghum. It is therefore
    postulated that the ethylene dibromide acts on the formation or
    release of pituitary follicle stimulating hormone (Olomucki, 1957).

    A group of laying hens that was fed for 23 days with oats fumigated
    with ethylene dibromide several months previously showed some
    diminution of egg size compared with controls and when the grain had

    been treated the day before with Dowfume EB5 (containing 63.6 per cent
    carbon tetrachloride, 29.2 per cent ethylene dichloride and 7.2 per
    cent ethylene dibromide by weight) at 10 times the recommended dose,
    egg output and size declined rapidly in 10 days (Bierer & Vickers,

    Five groups, each of 16 pullets six months of age, were fed for 19
    days on:

    (i)    a protein laying ration, plus non-fumigated oats;

    (ii)   a protein laying ration, plus oats fumigated at the rate of 1.1 

    (iii)  a protein laying ration, plus oats fumigated at the rate of 3.3 

    (iv)   a commercial, all-mash ration, non-fumigated;

    (v)    a commercial, all-mash ration, the corn component of which had
           been fumigated previously at the rate of 1.1 ml/kg.

    The fumigant was a commercial mixture containing carbon tetrachloride
    64 per cent, ethylene dichloride 29 per cent and ethylene dibromide 7
    per cent. After fumigation the grain was allowed to air for two weeks.
    In those groups which received fumigated oats, egg size was seriously
    and significantly depressed, more so at the heavy dosage (when
    production was totally arrested) than at the lower dosage. A smaller
    but still significant depression of egg production was observed in the
    birds on the mash ration containing fumigated oats subsequently ground
    (Caylor & Laurent, 1960).


    When ethylene dibromide is used as a fumigant on food products a small
    amount of the compound may be converted to inorganic bromide by
    reaction with the foodstuff, but the major part remains adsorbed to
    the food in the unchanged form for some time.

    There is little reason to believe that the limited changes brought
    about chemically in the foodstuff by the action of ethylene dibromide
    on its components are responsible for any significant changes in its
    food value or for the production of any toxic substances.

    The deleterious effects observed experimentally with the feeding of
    fumigated rations to animals can be attributed, in all probability, to
    the residue of unchanged ethylene dibromide.

    The feeding studies carried out with ethylene dibromide, either fed
    directly or as a residue after fumigation, were principally done with
    poultry. This species seems particularly sensitive to the compound,

    since experiments on other animals, though limited, give little
    indication of toxicity. This action of ethylene dibromide on poultry
    may be due to interference with the hormonal balance.


    The minimum pharmacologically effective dosage in the human adult has
    been generally stated to be about 900 mg of KBr, equivalent to 600 mg
    of bromide ion daily. Assuming an adult weight of 60 kg, this dosage
    in man is about 10 mg/kg body-weight per day. In the light of
    available toxicological information obtained from the extensive
    therapeutic use in man, an acceptable daily intake for man of
    inorganic bromide from all food sources can be estimated.

    Estimate of acceptable daily intake for man of inorganic bromide

         0-1.0 mg/kg body-weight.

    On the available toxicological evidence, ethylene dibromide should be
    used for fumigation of food only on the condition that no residue of
    the unchanged compound will reach the consumer.

    Further work required for unchanged ethylene dibromide

    Further investigation of the effect of processing and cooking on
    residual ethylene dibromide in food.

    Short-term feeding studies in two mammalian species.

    Reproduction studies for three generations in at least two species.

    Results of the above work should be made available not later than
    three years after the publication of this report, when a re-evaluation
    of this compound will be made.


    Use pattern

    (a) Pre-harvest treatment

    Ethylene dibromide has a fairly high phytotoxicity. Its pre-harvest
    uses are confined to the treatment of soil before planting and to
    certain, usually dormant, plants for quarantine purposes. Any residues
    in food derived from such uses are likely to be minute and to occur as
    inorganic bromide.

    (b) Post-harvest treatment

    Ethylene dibromide is rarely used by itself. In a few countries it is
    so used for stacked cereals in bags. It is used in many countries for
    treating cereals, in mixtures with other liquid fumigants such an
    ethylene dichloride and carbon tetrachloride. These mixtures are

    usually applied to the surface of grain standing in bins or in holds
    of ships. Ethylene dibromide is readily sorbed by individual grains
    but, when poured on to grain it only penetrates a short distance. This
    also pertains even when mixtures are used in this manner, since
    separation of the constituents rapidly occurs (Berck, 1961; Berck &
    Solomon, 1962). This chemical therefore is used sometimes in liquid
    grain fumigants to ensure an adequate treatment of the surface layers,
    where the highest concentrations of insects are frequently found. When
    such mixtures are applied to the surface of a bulk, the proportion of
    the grain present which comes into contact with ethylene dibromide is
    usually quite small; furthermore, samples carefully taken from surface
    layers may have high residues whereas the main volume of grain present
    may not contain any fumigant. Mixtures are also used for the "spot
    fumigation" of individual items of machinery in milling plants.

    Ethylene dibromide is used on fresh fruit such as citrus, mainly as a
    quarantine measure against fruit fly. It is also sometimes used in a
    mixture with other fumigants, such as ethylene dichloride and carbon
    disulfide, on some other stored foods, such as pulses, and on various
    non-edible products. Fresh fruits may sorb some fumigant but with
    thick-skinned fruits such as citrus, these residues appear to be
    concentrated in the skins and penetration into the flesh of undamaged
    fruit appears to be small (Page & Blackith, 1956; Grierson & Hayward,
    1959; Coggiola & Huelin, 1964). Ethylene dibromide is also used for
    the fumigation of empty wagons.


    There are no tolerances for residues of unchanged ethylene dibromide.

    For inorganic bromide derived from fumigation there are tolerances of
    50 ppm in Brazil, Germany, India, the Netherlands, USA and Canada;
    there is a tolerance of 20 ppm in Czechoslovakia and New Zealand.

    Residues resulting from supervised trials

    The fumigant is sorbed very readily by cereals or cereal products
    during exposure period (Whitney & Kenaga, 1960; Heuser, 1964).
    Although nearly all of the fumigant taken up by the cereals appears to
    be physically sorbed and can be removed by airing, this is a very slow
    process by comparison with the removal of methyl bromide. Therefore
    there is a greater chance of unchanged fumigant remaining in food.

    When whole or ground wheat was fumigated experimentally at 8 g/m3 or
    16 g/m3 for 24 hours, more than 95 per cent of the ethylene
    dibromide could be recovered unchanged after 10 days; most of it was
    retained in the bran. Very little was converted to ionized bromide.
    Residues in milled samples after 35 days amounted to 4.5 ppm
    (Sinclair, Lindgren & Forbes, 1962). The very small proportion of the
    sorbed fumigant which combines appears to react mainly with the
    protein, or to change into inorganic bromide. This may result in a
    very slight change in the nature of the food protein itself, probably
    in its methionine component (Bridges, 1956).

    Coggiola & Heulin (1964) found the uptake of ethylene dibromide by
    treated oranges during treatment to increase with the period of
    storage of the fruit before fumigation. These authors also found that
    the losses of residue after treatment were greater with longer periods
    of pre-fumigation storage. One hundred and fifty gram oranges stored
    one to 25 days before fumigation absorbed from approximately 30 mg to
    35 mg (respectively) of ethylene dibromide per orange; subsequent loss
    of residue varied from about zero up to over 5 mg per orange after 30

    Getzendaner (1965) has investigated the residues of various mixed
    commercial fumigants based on ethylene dibromide in milled wheat
    products following spot-fumigant treatment of mill machinery. For a
    mixture of 59 per cent ethylene dibromide with 32 per cent carbon
    tetrachloride and nine per cent ethylene dichloride residues up to
    1000 ppm of organic bromide were found immediately following
    fumigation, but giving way to pre-fumigation levels (10-20 ppm total
    Br. 0-4 ppm organic Br.) fairly quickly. 20.4 per cent ethylene
    dibromide with 57 per cent carbon tetrachloride, 19.6 per cent
    ethylene dichloride gave somewhat lower bromide residues in the flour
    immediately after fumigation (200-300 ppm) falling quickly to
    pre-fumigation levels. Getzandaner (1965a) has also shown that poultry
    meat and eggs from hens fed diets containing up to 400 ppm bromide
    residues contain at equilibrium no more bromide residue than the diet.
    Bär (1964) showed that the feeding of fumigated cereals to cattle may
    lead to bromide residues in milk. Lynn (1963) found 20 ppm bromide in
    milk from cows fed a diet containing 43 ppm bromide.

    Berck (1965) has shown that under laboratory conditions the uptake of
    ethylene dibromide by wheat increases significantly with increase of
    moisture content from nine to 18.5 per cent; the same is true of
    ethylene dichloride but not of carbon tetrachloride. Comminution, as
    in milling, greatly increased the uptake. The chromatographic
    characteristics of 51 different cereals (or cereal products) towards
    mixed fumigants was also studied.

    Residues in food moving in commerce

    Duggan, Barry & Johnson (1966) reported total bromide levels of up to
    200 ppm in various defined food groups in total diet studies; the
    findings are summarized in Table I; but Heywood (1966) pointed out
    that not all of this bromide could be attributed to the use of
    fumigants since some occurs naturally in food.

    Foods                 Bromide range ppm      Number of samples
                                                 (out of 18)

    Dairy products        1.1 to 31.7                17

    Meat, fish            2.3 to 35.5                16

    TABLE 1. (cont'd)
    Foods                 Bromide range ppm      Number of samples
                                                 (out of 18)

    Grain                 4.4 to 111.0               17

    Potatoes              1.5 to 38.0                15

    Leafy vegetables      1.1 to 16.3                16

    Legumes               0.9 to 17.9                14

    Root vegetables       2.6 to 22.1                14

    Salad fruit           1.7 to 18.9                15

    Fruit                 0.7 to 31.4                12

    Oils, fats            1.1 to 261.0               16

    Sugar etc.            0.7 to 55.1                18

    Beverages             0.9 to 17.0                10

    An examination of 227 grain shipments to the Netherlands from all
    parts of the world in the period May 1964 to September 1961, showed
    inorganic bromide residues to be present in only 12 of these. Methods
    for the measurement of unchanged residues of ethylene dibromide are
    still being developed but preliminary observations on these shipment:
    showed unchanged ethylene dibromide to be present in one sample only
    (approximately 0.5 ppm).

    Although results are available from a number of pilot scale trials
    using commercial sale, handling and milling facilities (e.g. Lynn &
    Vorkes, 1957), very few data are available for commercially marketed
    cereals. Because of the manner in which the fumigant separates out
    when liquid fumigant mixtures are applied to the surfaces of grain
    standing in tins, measurable residues may only be found in the surface
    layer of the grain present: reports on residues found should therefore
    give details of the sampling method used.

    Fate of residues

    In plants and animals

    Little work appears to have been done on mechanisms by which ethylene
    dibromide produces toxic effects in plants and animals or on its fate
    when it has been absorbed. Residues of "apparent organo bromine

    compounds" reported by Gunther & Spenger (1966) were said to be due
    to the extraction of inorganic bromide by organic solvents in the
    presence of lecithin-type complexing agents present in vegetable
    tissues and the results (some of which are reproduced on page 121)
    probably reflect naturally occurring bromide in the various plants.

    In storage and processing

    Ethylene dibromide in wheat and wheat products is very resistant to
    dispersion by airing. Toxic effects on animals, particularly poultry,
    have resulted from consumption of food treated with this fumigant.
    These effects have usually followed feeding relatively soon after a
    treatment, bearing in mind the low rate of dispersion, and seem to be
    attributable to the presence of free ethylene dibromide. Almost all
    traces of ethylene dibromide appear to be lost by volatilization on
    heating as in baking (Senger & Mapes, 1957; Munsey, Mills & Klein,
    1957) but according to Bridges (1956) some breaks down to produce
    minute amounts of ethylene glycol which may react with the - SCH3 of
    the methionine in the wheat protein. The joint meeting of the FAO
    Committee on Pesticides in Agriculture and the WHO Expert Committee on
    Pesticide Residues (FAO/WHO, 1965) recommended that the effects of
    processing and particularly of cooking on the residues of ethylene
    dibromide in foods should be further investigated.

    Methods of residue analysis

    In sampling cereals for ethylene dibromide residue analysis, special
    attention must be paid to the extent to which the sample is
    representative of the bulk. Analysis of residues of ethylene dibromide
    in cereals, which are mostly present in the unchanged organic form, is
    usually based on the measurement of total organic and inorganic
    bromine, measured as bromide. For this, there are a number of well
    established methods based on the hydrolysis of ethylene dibromide with
    alkali to inorganic bromide, evaporation to dryness of the resultant
    mixture, removal of the organic matter by controlled ignition,
    extraction by dilute acid of the total bromide from the resultant ash,
    oxidation to bromate, the liberation by this of iodine from acid
    potassium iodide and the determination of the iodine titrimetrically
    with sodium thiosulfate (Mapes & Shrader, 1957; Heuser, 1961). The
    inorganic bromide residue alone may similarly be determined after
    removal of unchanged ethylene dibromide by aeration (Heuser, 1961.).
    Unchanged ethylene dibromide can then be calculated by difference. The
    limit of detection by this procedure is of the order of 1 ppm as
    bromide. Berck (1965) has published details of a gas chromatograph
    method for the detection and measurement of some 34 fumigant vapours,
    including ethylene dibromide. This general technique has recently been
    applied by Bielorai & Alumot (1965) to the measurement of residues of
    unchanged ethylene dibromide in experimentally fumigated animal feeds:
    its application to the determination of such residues in commercially
    fumigated foods is most desirable.

    The method for the determination of total and inorganic bromide
    derived from the use of ethylene dibromide on cereals based on that of
    Heuser (1961) is in the meantime suggested. Details are set out in
    Appendix A(1) of the (Extract) Report of the Second Session of the FAO
    Working Party on Pesticide Residues (PL/1965/12).


    On the basis of an acceptable daily intake of 1 mg/kg of inorganic
    bromide, a tolerance of 50 ppm for inorganic bromide residues in
    cereals and flour resulting from the use of ethylene dibromide and
    other fumigants can be recommended. This recommendation is also
    consistent with tolerance for dried eggs, processed herbs and spices
    at 400 ppm, various fresh fruits at 200 ppm, citrus at 230 ppm,
    raisins and dates at 100 ppm, and various other fruits, including
    certain dried fruits, at from 50 ppm to 20 ppm, which are residues
    which have been found after responsible use of the fumigant and which
    are as follows:

            Dried eggs, spices, herbs       400
            Cereals                          50
            Dried figs                      250
            Avocados                         75
            Dried raisins, dates            100
            Dried peaches                    50
            Dried prunes                     20
            Other dried fruits               30
            Citrus, strawberries             30
            Other fresh fruit                20

    In the absence of a recommendation for an acceptable daily intake for
    unchanged ethylene dibromide and as there appear to be no very strong
    practical or economic reasons for accepting such residues, no
    recommendations are made for a tolerance for residues of this

    The joint FAO/WHO Expert Committee on Food Additives (FAO/WHO, 1964),
    when considering the presence of bromide in bread due to the addition
    of bromate conditioner to flour, gave estimates of acceptable levels
    for treatment for flour to be consumed by man as:

    Unconditional                           0-20 ppm
    Conditional (for special purposes)     20-75 ppm

    The recommendation of 50 ppm in cereals and flour resulting from
    fumigation is in accordance with this estimate because cereals and
    cereal products are treated with the fumigant to deal with specific
    outbreaks and not as a routine, during milling or other manufacture,
    as are flour conditioners.

    Further work or information

    Further data are needed on the occurrence of residues in raw and
    processed foods after use of the fumigant under practical conditions.
    Measurement of residues in foods resulting from the use of mixed
    fumigants, including ethylene dibromide, is desirable.

    Gas-liquid chromatographic methods should be particularly useful for
    the determination of residues of unchanged ethylene dibromide in
    cereals. The method offers the prospect of increased sensitivity and a
    means for the simultaneous determination of other fumigants which may
    be present. It should now be possible to develop a rapid, selective,
    quantitative technique for traces of halogenated fumigants, together
    with suitable desorption techniques, and to combine these into a
    single method for the determination of residues in treated foods and
    the Working Party recommends accordingly. Figures for the levels of
    any unchanged ethylene dibromide in commercially treated materials
    particularly those which have a relatively high oil or fat content,
    would also be useful.


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    See Also:
       Toxicological Abbreviations
       Ethylene dibromide (ICSC)
       Ethylene dibromide (FAO Meeting Report PL/1965/10/2)
       Ethylene dibromide (FAO/PL:1967/M/11/1)
       Ethylene dibromide (FAO/PL:1968/M/9/1)
       Ethylene Dibromide (IARC Summary & Evaluation, Volume 71, 1999)