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    DICHLOFLUANID        JMPR 1974

    Explanation

          In 1969 the Joint Meeting evaluated dichlofluanid in the light of
    the information then available. No ADI could be established and no
    tolerances were recommended.

          The Meeting listed 10 items on which further work or information
    was required before tolerances could be recommended (FAO/WHO, 1970).

          New information has been received on toxicology, composition of
    the technical product, rates and frequencies of application,
    pre-harvest intervals, residues and the influence of residues on the
    fermentation of must. This information is summarized in the following
    monograph addendum.

    IDENTITY

    Synonyms

          Elvaron(R)

    Other relevant chemical and physical properties

          The purity of the technical material is at least 96%, and the
    maximum levels of the impurities are as shown in Table 1.

          In practice, the average content of dichlofluanid in the
    technical product is approximately 98%, so that the by-products
    normally amount to only about half of the amounts given in Table 1.

    TABLE 1. Maximum levels of impurities in dichlofluanid

                                                                Maximum
    Impurity                                                    content, %

    N,N-dimethyl-N-phenylsulphamide (DMSA)                      1.5

    N,N-dimethylbenzylamine (C6H5-CH2-N(CH3)2)                  1.0

    "difluoro-dichlofluanid" (R-CF2Cl)                          0.3

    chlorides (expressed as HCl)                                0.3

    up to 6 unidentified trace impurities, total                1.0

    EVALUATION FOR ACCEPTABLE DAILY INTAKE

    BIOCHEMICAL ASPECTS

    Absorption, distribution, biotransformation and excretion

          No information is available on the rate of absorption or on the
    distribution of dichlofluanid in the animal body. Studies in rats
    indicate that the absorption is small. Within 72 hours, 45-92 percent
    of the administered dose could be isolated from the faeces. Most of
    the dichlofluanid found was in an unchanged form; approximately 12
    percent was isolated as N,N-dimethyl-N'-phenylsulphamide. After oral
    administration of dichlofluanid the unchanged form of the compound was
    not detectable in the serum or urine; however when either
    dichlofluanid or its metabolite dimethylphenylsulphamide was
    administered orally, four metabolites could be isolated from the
    urine. The structures of these compounds have now been determined and
    are given below:


    CHEMICAL STRUCTURE 4


    N,N-dimethyl-N'-phenylsulphamide (I)


    CHEMICAL STRUCTURE 5


    N-methyl-N'-phenylsulphamide (II)


    CHEMICAL STRUCTURE 6

    N,N-dimethyl-N'-(4-hydroxyphenyl) sulphamide (III)

    CHEMICAL STRUCTURE 7

    N-methyl-N'-(4-hydroxyphenyl)-sulphamide (IV)

          All the metabolites are excreted in the free form and the
    metabolites III and IV are also excreted as glucuronides. Metabolites
    I and II can also be found in serum. The metabolites found in urine
    and serum after oral administration of dichlofluanid are the same as
    those found after oral administration of dimethylphenylsulphamide.
    From these studies it is evident that dichlofluanid is absorbed only
    very slightly from the G.I. tract (Eben and Kimmerle, 1968; Anon.,
    1969).

          In the studies on the metabolism of dichlofluanid there is no
    information on the fate of the dichlorofluoromethylthio portion of the
    molecule, and it is not known if the fluorine atom appears ultimately
    as fluoride ion. However, alkaline hydrolysis of dichlofluanid in
    methanol solution is reported to yield
    N,N-dimethyl-N'-phenylsulphamide and fluorodichloromethanethiol
    (Cl2FCSH) which is susceptible to oxidation. No information is given
    on the nature of the oxidation products (Nangniot et al., 1967).

    TOXICOLOGICAL STUDIES

    Special studies on the metabolite, N,N-dimethyl-N'-phenylsulphamide

    Rat

          Groups of 30 rats (15 of each sex) were fed 0, 1000, 3000, and
    10 000 ppm of dimethylphenylsulphamide in their diets for four months.
    Mortality, food consumption, growth, haematology, urinalysis gross and
    microscopic pathology (10 animals from each group) were closely
    comparable in the experimental and the control groups. Also the final
    average body and organ weights were comparable in the different
    groups, except in the 10 000 ppm group where the female rats displayed
    a decrease in the adrenal weight, and in both sexes an increase in the
    liver weight was found. Although no histopathological changes in the
    liver and the kidney of these rats were reported, such changes might
    be camouflaged by the histological changes due to infections (Lorke,
    1965).

    Special studies on mutagenicity

    Mouse

          A dominant-lethal-test was carried out by administering 500 mg
    dichlofluanid/kg bodyweight to twenty male mice. Untreated female mice
    mated with three males did not show any difference in the early and
    late resorptions, the number of corpora lutea or living and dead
    foetuses when compared to those mated with control males. This study
    at one relatively high dose level, did not reveal any dominant lethal
    mutagenic effect of dichlofluanid (Machemer, 1974b).

    Special studies on reproduction

    Rat

          Groups of rats, each comprising 10 males and 20 females received
    dietary levels of 0, 150, 500, 1500 and 4500 ppm of dichlofluanid
    (90.2% pure; 8% inert material; 0.3% DMSA) for a period extending over
    three generations. Two litters per generation were studied. No
    information is available on the selection of animals from one
    generation to the next. None of the dosed groups indicated any
    abnormal effects on fertility throughout all three generations. The
    groups fed 150, 500 and 1500 ppm displayed no abnormal effects
    compared to controls with respect to group averages of birth weight,
    litter-size and percentage of survival to weaning. Data for still
    births and resorptions are not given in the report of this study. Only
    in the group given 4500 ppm, the body weights of the young animals
    were lower both at birth and at weaning in all generations except the
    F1a at birth. The lactation index was also slightly reduced in one of
    the six matings at this level. Although the experiment was not
    designed to study teratological effects it may be noted that no
    malformations were seen at any dose level (Löser, 1969a).

          An additional study of the histology of organs of some animals of
    the F3b generation showed no dose related effects of dichlofluanid
    (Vince and Spicer, 1971).

          Groups of 20-23 pregnant female rats were administered
    dichlofluanid by stomach tube, on days 6 to 15 of gestation, at doses
    of 30, 100, and 300 mg/kg/day. Symptoms of toxicity were observed in
    all test groups and body-weight gains were decreased compared to the
    controls both during the ten days treatment and during the total
    gestation period. The effect was similar in all test groups and not
    dose-dependent. The number of implantations, resorptions and foetuses
    were within normal limits in all groups, whereas the average weight of
    the placenta and the foetuses were slightly decreased in the groups
    administered 300 mg/kg, possibly owing to the general toxic effect on
    the maternal animals. In this study comprising only doses with
    maternal toxicity no indication of teratogenic potential of
    dichlofluanid was found (Machemer, 1974a).

    Acute toxicity

    TABLE 2.  Acute toxicity of dichlofluanid

                                                                            

                                           LD50mg/kg
    Animal                     Route       body-weight      References
                                                                            

    Mouse        (F)           i.p.        7.8              DuBois and
                                                            Raymund, 1963

    Mouse        (M)           i.p.        6.0              Dubois and
                                                            Raymund, 1963

    Chicken                    oral        >1000            Dubois, 1963

    Rat          (M)           i.p.        15               Dubois and
                                                            Raymund, 1963

    Rat          (F)           i.p.        15               Anon., 1962

                                                            DuBois and
                                                            Raymund, 1963

    Rat          (M)           oral        500-1000         DuBois and
                                                            Raymund, 1963

    Rat          (F)           oral        525              Anon., 1962

                                                            DuBois and
                                                            Raymund, 1963

    Guinea-pig   (M)           i.p.        35               DuBois and
                                                            Raymund, 1963

    Guinea-pig   (M)           oral        250              DuBois and
                                                            Raymund, 1963
                                                                            


          The symptoms of poisoning were non-typical and consisted mainly
    in a decrease in activity which began several hours after
    administration of the compound. With doses around the LD50, death
    occurred in one to four days (DuBois and Raymund, 1963).

    Short-term studies

    Rat

          Groups of 30 rats (15 of each sex) were fed 0, 30, 100, 300,
    1000, 3000, and 10 000 ppm of dichlofluanid in their diets for four
    months. Food consumption, growth, haematology, urinalysis, mortality,
    and gross and microscopic pathology were closely comparable in the 30
    to 3000 ppm experimental groups and the control groups. The same was
    the case with final average body and organ weights except in the 3000
    ppm group where the male rats displayed reduction of heart weights and
    the female rats an increase in liver weights. In the rats fed 20 000
    ppm there was a deterioration of general condition, decreased food
    intake, smaller weight gains and higher mortality than the control
    group. The male rats showed a decrease in heart weights, while in both
    sexes there was a decrease in the weight of adrenal glands and an
    increase in liver weights. Histopathological changes were found in the
    liver (vacuolization, inflation in size and shrunken nuclei of some
    cells), kidneys (increased protein precipitation in the proximal
    tubuli) and the spleen (reduction of the lymphatic tissue) (Anon.,
    1964).

    Dog

          Groups of four dogs (two of each sex) were fed 0, 500, 1500, and
    4500 ppm of dichlofluanid in their diets for four months. Behaviour,
    food consumption, body-weight changes, mortality, function tests of
    the liver and kidney, urinalysis, gross pathology and final average
    body weights and average organ weights were closely comparable in the
    500 ppm and the control group. The same was the case with the male
    dogs of the 1500 ppm group while in the females changes in
    liver-function tests and decrease in body weight pointed at an 
    impaired liver-function. Three of the four dogs fed 4500 ppm of
    dichlofluanid died. Before death these dogs and also the surviving
    animal displayed signs of impaired liver and kidney function (Lorke
    and Löser, 1966).

          Groups of 4 male and 4 female dogs were fed 0, 100, 300, 1000 and
    3000 ppm dichlofluanid (97.7% pure) in their diet for 2 years.
    Behaviour, body weights, food consumption, mortality, blood and urine
    analyses, liver and kidney function tests and absolute and relative
    organ weights for the groups fed 100, 300 and 1000 ppm did not differ
    from the values of these parameters in the controls. In the groups fed
    3000 ppm increased mortality, reduced body weight and food consumption
    and decreased absolute and relative testis weight were found.
    Microscopic examination of the organs revealed an increase in
    interstitial tissue of the testis and vacuolation and degeneration of
    the adrenal cortex of the 3000 ppm group as the only dose-dependent
    pathological changes (Löser, 1969b; Mawdesley-Thomas et al., 1971).

    Long-term studies

    Rat

          Groups of 80 rats (40 of each sex) were fed 0 (two groups), 150,
    500, 1500 and 4500 ppm of dichlofluanid in their diets for two years.
    Behaviour, food consumption and mortality of the test groups did not
    differ from the parameters of the control groups. The same was the
    case with haematology (at 4 and 24 months) and liver function and
    urine examination (at 24 months). Gross pathology of animals which
    died during the experiment, and of sacrificed test rats at the end of
    the study, did not reveal any changes that might be caused by
    dichlofluanid, but no histological examination of the organs is
    recorded. Female rats in the 4500 ppm group showed decreased weight
    gain and relative kidney weights, while both sexes showed elevated
    relative liver weights, but normal enzyme function tests (Löser,
    1968).

          Histological examinations were made on tissues of five male and
    five female rats at the end of the study and on animals that died
    during the experiment. No significant pathological changes were found
    at dietary levels including the 4 500 ppm level. A separate study of
    all tumours found at autopsy was performed, and there was no
    indication that dichlofluanid produced an increased tumour incidence
    (Mawdesley-Thomas, 1969).

    Comments

          In a 2-year study in dogs a 1 000 ppm level of dichlofluanid in
    the diet appears to have no untoward effect, although no data are
    given on the individual animals.

          A long term study in rats indicated 1 500 ppm in the feed as the
    level without apparent toxic effect. There was no indication of
    teratogenic effect when female rats were administered a dose as high
    as 300 mg/kg bw. Reproduction parameters were not affected in a
    3-generation feeding test with levels up to 1500 ppm dichlofluanid.
    However, there is a lack of information on absorption, distribution,
    excretion and pharmacokinetics on the dichloro-fluoromethylthio moiety
    which may produce some carbon-fluoride compounds resistant to
    cleavage. The Meeting allocated a temporary acceptable daily intake
    for man based on the no-effect level in the dog, the more sensitive
    species.

    TOXICOLOGICAL EVALUATION

    Level causing no toxicological effect

          Rat: 1 500 ppm in diet equivalent to 75 mg/kg bw.
          Dog: 1 000 ppm, in diet equivalent to 25 mg/kg bw.

    

    ESTIMATE OF TEMPORARY ACCEPTABLE DAILY INTAKE FOR MAN

          0 - 0.3 mg/kg bw

    RESIDUES IN FOOD AND THEIR EVALUATION

    USE PATTERN

    Pre-harvest treatments

          Uses (Table 3) are additional to those listed in FAO/WHO, 1970.
    They are all recommendations of the Federal Republic of Germany (FRG).

    TABLE 3.  Pre-harvest treatments with dichlofluanid recommended
              in the Federal Republic of Germany
                                                                            
                                  Number of           Rate of application
    Crop                          applications        (kg a.i./ha)
                                                                            
    Pome fruits                   4 - 11              1.5

    Stone fruits

       plums                      4                   1.5 - 2.0

       cherries                   3 - 8               2.0

       peaches                    3                   1.5

    Soft fruits

       strawberries               1 - 6               2.5 - 3.0

       red and black
       currants                   4 - 7               2.0 - 2.4

       raspberries                1 -4                2.0 - 2.4

    Grapes                        2 - 10              1.9 - 2.5

    Vegetables

       cucumbers                  1 - 4               1.5 - 2.0

       tomatoes                   1 - 3               1.5 - 2.0

       lettuce (field grown
       and under glass)           2 - 4               0.6 - 1.0

    Beans                         1 - 3               1.0

    Hops                          4 - 17              2.0
                                                                            



    
    TABLE 4.  Residues of dichlofluanid (I) and DSMA (II) resulting from supervised trials.

                                                                                                                                               

    Crop                  Application                 Residue, mg/kg, after interval (days)
    (Country)             Rate
    Reference1            kg/ha    No.    Compound    0-3          4-7          8-14         15-21        22-28        29-35        36-42
                                                                                                                                               

    Apples (FRG)          1.1-     4-     I            0.7-3.4      0.2-2.8     <0.1-2.2     <0.1         <0.1-0.4                  <0.1
                          1.25     11     II           0.3-1.1     <0.1-1.1     <0.1-1.1     <0.1-0.4     <0.1                      <0.1

    Beans green incl.     1.0-     1-     I                                      0-1-0.3      0.1-0.2     <0.1-0.2                   0.1
    pods (FRG; UK)        3.4      3      II                                     0.1-0.2      0.1-0.2     <0.1                       0.1

    Cherries (morello)    1.1      6-8    I            0.6-2.5     <0.1-0.4     <0.1         <0.1
    (FRG)                                 II           0.2-1.2     <0.1-0.2     <0.1         <0.1

    Cucumbers (outdoor);  1.5      3      I           <0.1-0.2                  <0.1-0.2     <0.1
    (FRG)                                 II           0.2-0.5                  <0.1-0.3     <0.1

    Cucumbers (under      1.5-     1-     I            0.3-3.6     0.2-0.3      <0.1         <0.1
    glass); (FRG;         6.3      3      II           0.1-0.5     <0.1         <0.1         <0.1
    Netherland

    Currants, black       2.5/?    2-7    I            2.4-48       2.0-16       2.0-6.3      0.9-5.1      0.5-1.4      0.2
    (Denmark2/UK)                         II           0.5-3.2      1.1-1.5      0.5-1.0      0.4-0.7      0.2-0.3      0.3

    Currants, red (FRG;   0.2-     3-5    I            5-39         3-23         2-46         1-40         0.3-3.2
    Netherland3)          2.5             II           1.9-6.9      1.1-3.0      0.4-16       0.5-3.0      0.2-0.4

    Gooseberries (UK)              5      I                                      0.3
                                          II                                     0.2

    Wheat, barley
    (France)              1.3      1-2    I                                                                                         <0.1

    Grapes (FRG4)         0.6-     2-     I            0.2-48       0.2-33       0.7-10.6    <0.1-21      <0.1-16       1.3-11      <0.1-12
                          3.6      10     II          <0.1-0.8     <0.1-3.8     <0.1-3.6     <0.1-2.7     <0.1-2.6      0.3-3.4     <0.1-3.8

    TABLE 4.  (Cont'd.)

                                                                                                                                               

    Crop                  Application                 Residue, mg/kg, after interval (days)
    (Country)             Rate
    Reference1            kg/ha    No.    Compound    0-3          4-7          8-14         15-21        22-28        29-35        36-42
                                                                                                                                               

    Hops (FRG)            2.0      9-     I            69           6-140        5            0.3-0.7
                                   17     II           14           3-11

    Lettuce (outdoor)     0.6      1-4    I            7.8-30       0.2-2.7      0.1-0.2     <0.1-0.3     <0.1                      <0.1
                                          II           2.1-5.8      0.2-2.3     <0.1         <0.1-0.3      0.1-0.2                  <0.1

    Lettuce
    (under glass)         0.6-     1-6    I            4.7-55       0.3-8.3     <0.1-1.9     <0.1-0.3     <0.1-5.6                  0.6-2.2
    (FRG; UK)             3.4             II           3.0-13       0.2-2.8     <0.1-2.5     <0.1         <0.1-1.6

    Onions (UK)           1.1      5      I                                                                                         (100 days)
                                                                                                                                    <0.1

    Peaches (FRG)         1.1      3      I            2.2-2.9      0-7-1.2      0.3-1.6
                                          II           0.5-0.8      0.2-1.4      0.5-0.9

    Raspberries (FRG;     2.0-     1-4    I            21-32        6-14        <0.1-10.5    <0.1-4.7      1.7
    Netherland            2.3             II           1.0-6.2      0.7-6.4      0.6-4.0      0.8-1.0      0.9

    Strawberries
    (outdoor)             0.8-     1-4    I            0.3-27       0.1-5.0     <0.1-6.3     <0.1-2.4     <0.1-1.6     <0.1-1.7      0.7-1.7
    (FRG: Netherland      3.8             II           0.3-8.8      0.5-1.8     <0.1-3.7      0.2-2.4      0.3-1.9     <0.1-0.3      0.1
    N.Z.6; Switzerland;
    UK)

    (Australia7)          1.4-     3      I + II                    1.2-15.5     0.4-8.4     <0.1-3.2     <0.1-0.6      0.6         <0.1
                          2.1             expr. as I

    Strawberries
    (under glass)         1.2-     3-6    I            4-9          0.5-3.5      0.4-7.1      2.2-4.1      1.7-3.5      1.5-4.7     <0.1-3.6
    (Netherland           9.5
                          (total)

    TABLE 4.  (Cont'd.)

                                                                                                                                               

    Crop                  Application                 Residue, mg/kg, after interval (days)
    (Country)             Rate
    Reference1            kg/ha    No.    Compound    0-3          4-7          8-14         15-21        22-28        29-35        36-42
                                                                                                                                               

    Tomatoes (FRG)                 1-3    I           <0.1         <0.1-0.2
                                          II          <0.1-0.2     <0.1-0.9
                                                                                                                                               

    1 Bayer AG (1964-1973) unless otherwise stated
    2 Government Plant Pathology Institute, Lyngby; National Pood Institute, Gladsaxe, April 20, 1970
    3 Keuringsdienst van Waren Amsterdam (1967a, 1969a)
    4 Lemperle et al. (1969, 1970); Bayer (1964-1973)
    5 Keuringsdienst van Waren Amsterdam (1965)
    6 Brewerton and Gibbs (1968)
    7 Tate at al. (1973)
    8 Keuringsdienst van Waren Amsterdam (1969b)
    


    RESIDUES RESULTING FROM SUPERVISED TRIALS

          Table 4 shows the residues of dichlofluanid and its metabolite
    dimethylphenylsulphamide (DMSA) found in crops after recommended
    applications in supervised trials. The analyses were carried out by
    Farbenfabriken Bayer AG and various institutes.

    FATE OF RESIDUES

    In storage and processing

          Additional information on the fate of residues during processing,
    including wine making, and on the effect of residues on fermentation,
    has been received since the previous evaluation.

          Analyses of commercial deep-frozen strawberries from two
    factories (3 samples from each) showed residues of 0.26 - 0.36 and
    0.06 - 0.12 mg/kg dichlofluanid. In canned strawberries residues were
    <0.02 mg/kg dichlofluanid and DMSA could not be found (Grevenstuk,
    1971).

          Strawberries were treated with dichlofluanid at flowering and
    analysed with and without caps (Government Plant Pathology Institute,
    Lyngby, Denmark, 1969). Most of the dichlofluanid residue was in the
    caps (Table 5).

        TABLE 5.  Residues of dichlofluanid and DMSA in strawberries with and
              without caps (Denmark)

                                                                                                

    Rate of                       Waiting                    Residues (mg/kg)
    application       No. of      Period        With cap                Without cap
    (kg.a.i./ha)   applications   (days)     Dichlofluanid   DMSA       Dichlofluanid    DMSA
                                                                                                

    1.9                  3          24           1.6         0.4             0.2         -

    (treated                        28           1.3         0.3             -           -
    at
    flowering)                      35           1.7         -               0.1         0.1

                                    42           0.7         -               0.1         0.1
                                                                                                
    
          Apples containing residues of about 0.8 mg/kg dichlofluanid plus
    0.2 mg/kg DMSA were cold pressed. The juice contained approx.
    0.2 mg/kg dichlofluanid and approx. 0.7 mg/kg DMSA. After heating the
    juice for 30 min. at 90°C the residues were 0.1 and 0.7 mg/kg
    respectively. Processing the apples to stewed fruit, baby food or
    jelly resulted in residues of approx. 0.05 mg/kg dichlofluanid and 0.3
    - 0.7 mg/kg DMSA (Maier-Bode, 1972). Residues of 2 mg/kg dichlofluanid
    plus 0.7 mg/kg DMSA on apples when stored at -18°C were stable for at
    least 6 months (Bayer, 1973).

          Several series of experiments were carried out to determine the
    effect of wine-making processes on residues in grapes, and the effect
    of dichlofluanid residues on fermentation.

          At residues of 1.5 - 10.8 mg/kg dichlofluanid and 0.02 - 0.1
    mg/kg DMSA on grapes the corresponding residues (mg/kg) in the marc
    were <0.05 - 8.0 and 0.1 - 1.4; in must (not clarified) <0.1 - 6.9
    and 0.1 - 1.3; in clarified must <0.05 - 0.2 and 0.3 - 0.8 and in
    wine <0.05 and 0.1 - 4.0 (Lemperle et al., 1970). There is no direct
    correlation between the residues on grapes and those in the processed
    products.

          In a further study residues up to 48 mg/kg dichlofluanid and 1.6
    mg/kg DMSA on grapes resulted in corresponding residues (mg/kg) up to
    6.9 and 4.5 mg/kg in must (not clarified), 0.4 and 2.3 mg/kg in
    clarified must and <0.05 and 4.0 mg/kg in wine (Lemperle et al.,
    1973).

          In other experiments (Lemperle and Kerner, 1969; Lemperle et al.,
    1970), grapes were treated two or three times at recommended
    concentrations. Residues of dichlofluanid and DMSA were determined in
    grapes harvested at intervals from 32 to 63 days after the last
    application, and in the wine made from them. Residues in the grapes
    (19 samples) ranged from 1.5 to 12 mg/kg dichlofluanid and 0.02 to 0.8
    mg/kg DMSA. In the wine, dichlofluanid could not be detected in any of
    the samples (limit of detection 0.02 - 0.05 mg/kg) and DMSA residues
    ranged from 0.1 to 4.0 mg/kg.

          Bayer (1967-1973) report trials in FRG in which grapes were
    treated at various rates with dichlofluanid. The results (Table 6)
    show residues of dichlofluanid from 0.02 - 5.4 mg/kg on the grapes
    which have been eliminated during processing to wine. DMSA residues
    are again carried through into the wine.

    TABLE 6.  Residues of dichlofluanid (I) and DMSA (II) on grapes
              and in wine produced therefrom

                                                                            

        Application     Days after             Residues, mg/kg*
       Rate             last              Grapes                Grapes
    kg a.i./ha    No.   treatment      I          II         I          II
                                                                            

    -             7     28             -          -          n.d.       -

    0.6           4     56             5.4        1.35       n.d.       -

    1.5 - 2.0     2     57             0.9        0.15       n.d.       -

    2.0 - 2.5     4     47             0.7        0.25       n.d.       0.2

    2.0 - 2.5     5     35             0.4        0.6        n.d.       0.25

    2.0 - 2.5     6     22             1.6        0.95       n.d.       0.4

    3.0 - 5.0     3     34             3.9        0.2        n.d.       n.d.

    2.0 - 4.0     4     59             -          -          n.d.       2.4

    3.6           2     60             -          -          n.d.       n.d.

    1.8           5     71             1.0        -          0.03       -

    0.8           8     54             0.02       -          n.d.       -
                                                                            

    * Limit of detection:   dichlofluanid 0.02 mg/kg;
                            DMSA 0.1 mg/kg.


          Fermentation experiments were conducted to determine the
    concentration of dichlofluanid required to inhibit yeast growth. At 1
    mg/l in must the fermentation was retarded and at > 1.5 mg/l it was
    inhibited. The inhibitory effect of dichlofluanid on yeast was
    abolished by cysteine, beta-mercapto-ethylamine and other
    SH-compounds. Dichlofluanid can therefore be regarded as an
    "SH-blocker." The inhibitory effect decreases with increasing pH. The
    activity of most enzymes depends on free SH-groups which are part of
    cysteine. Therefore the enzymes alcohol-dehydrogenase and
    glycerinaldehyde-3-phosphate-dehydrogenase which contain essential
    SH-groups were strongly inhibited in vitro and in vivo while maleic
    dehydrogenase was not affected. The fluorodichloromethylthio
    (-S-CCl2F)-part of the dichlofluanid molecule was shown to be the
    inhibiting group (Dittrich and Issinger, 1969).

          Retardation of the fermentation process can be overcome by
    pre-clarification of the grape juice (must) and addition of about 1%
    of a fermenting pure yeast strain preparation, or in the case of red
    grape juice, by the addition of 2.5% pure yeast while still in contact
    with the skins. In general the fermentation process seems to be less
    influenced by dichlofluanid than by folpet. No differences between
    wines originating from dichlofluanid-treated and non-treated vines
    could be observed either analytically or by tasting (Lemperle et al.,
    1970, 1973).

    RESIDUES IN FOOD IN COMMERCE OR AT CONSUMPTION

          Residues in 52 samples of red currants examined in the
    Netherlands were <0.1 mg/kg (Keuringsdienst van Waren, 1966). Of
    276 samples of strawberries, 256 contained <0.1 mg/kg
    dichlofluanid, 17 contained 0.1 - 1 mg/kg and 3 contained 1 mg/kg
    (Keuringsdienst van Waren, 1966).

          Results of food inspection reported in 1973 (Netherlands) are
    shown in Table 7.

    TABLE 7.  Residues of dichlofluanid found during food inspection.
              Netherlands, 1973

                                                                            

                                  Number of samples in range
    Range,                                          red
    mg/kg         blackberries    raspberries     currants     strawberries
                                                                            

     0 - 0.1           15              1             23           257

    >0.1-0.2           2                             7            111

    >0.2-0.5           7               3             11           215

    >0.5-1             5               3             8            168

    >1  - 2            4               2             5            134

    >2  - 5*           2               4             2            58

    >5  - 10           3                                          3

    >10                                                           1 (10.3
                                                                      mg/kg)

    Total              38              13            56           947
                                                                            

    * The tolerance was 5 mg/kg in all cases.

    METHODS OF RESIDUE ANALYSIS

          A multiresidue GLC method, with electron-capture detection
    (Becker, 1972/74), has been adopted by Deutsche Forschungsgemeinschaft
    (DFG, 1974) for the determination of dichlofluanid. The method in
    principle is based on the GLC method of Vogeler and Niessen (1967)
    already mentioned in FAO/WHO, 1970. For stewed apples, pears, plums,
    cherries, peaches, grapes, cabbage, carrots, green beans and lettuce
    the recovery at the 0.1 mg/kg level is > 80%. For leeks, parsley and
    celery the recovery is below 70%. The method is suitable for
    regulatory purposes for dichlofluanid residues in a number of crops,
    but further evaluation is desirable before it can be recommended
    without reservation. It does not determine DMSA.

          A similar analytical method has been elaborated, in which
    isopropanol-benzene extracts of the crop are cleaned-up with
    Nuchar-Attaclay. The method has been checked for many green
    vegetables, tomatoes and fruit. Interferences have sometimes been
    found for leek, onion, cabbage and carrots (Rijks Instituut voor de
    Volksgesondheid, Bilthoven, 1972).

    NATIONAL TOLERANCES REPORTED TO THE MEETING

          National tolerances and safety intervals reported to the Meeting
    are given in Table 8.

    TABLE 8.  National Tolerances for dichlofluanid reported
              to the Meeting

                                                                             

                                                 Safety
                                                 interval          Tolerance,
    Count          Crop                          (days)              mg/kg
                                                                             

    Australia      General                       14

    Austria        General                       14

    Belgium        Fruit, vegetables                               5.0
                   (excl. potatoes)

                   Strawberries (outdoor),
                   raspberries, mulberries        7

                   Strawberries
                   (under glass)                 14

                   Currants                      21

    Bulgaria       General                       14

    TABLE 8.  (Cont'd.)
                                                                             

                                                 Safety
                                                 interval          Tolerance,
    Count          Crop                          (days)              mg/kg
                                                                             

    Denmark        Fruit, including
                   strawberries                  14

    Finland        General                       14

                   Strawberries                                    (   4.0

                   Other fruit                   (proposed)        (   1.0

    France         General                        7

    Germany        Small fruit (berry            dichlofluanid     (  15.0
     (FRG)         fruit excluding               + DMSA            (
                   strawberries),                calculated as     (
                   grapes                        dichlofluanid     (
                                                                   (
                   Strawberries,                                   (
                   lettuce,                                        (  10.0
                                                                   (
                   all other fruit                                 (   5.0
                                                                   (
                   Beans, cucumbers,                               (
                   tomatoes                                        (   3.0
                                                                   (
                   Onions                                          (   1.0

                   Lettuce (field
                   grown)                        21

                   Lettuce (under
                   glass)                        28

                   Tomatoes (field
                   grown and under
                   glass)                         3

                   Cucumbers (field
                   grown and under
                   glass)                         3

                   Pome-fruit                     7

                   Stone-and small
                   fruit (excluding
                   strawberries)                 14

    TABLE 8.  (Cont'd.)
                                                                             

                                                 Safety
                                                 interval          Tolerance,
    Count          Crop                          (days)              mg/kg
                                                                             

    (Germany)      Strawberries                  10

                   Grapes                        35

    Italy          Grapes                        40

                   All other crops               20

    Japan          Cucumbers, tomatoes,
                   strawberries (under
                   glass)                         1

    Netherlands    General                                         5.0

                   Tomatoes, peppers,
                   cucumbers, melons              3

                   Strawberries (outdoor),
                   blackberries,
                   raspberries                    7

                   Strawberries                  14
                   (underglass)
                   Currants                      21

                   Kohlrabi                      14

    New Zealand    Fruit and vegetables                            5.0

                   General                       14

    Norway         General                        7

    Poland         Vegetables, fruit             14

                   Field grown crops
                   (excluding, potatoes,
                   beets)                        14

                   Grapes                        42

    Portugal       Strawberries                  14

    TABLE 8.  (Cont'd.)
                                                                             

                                                 Safety
                                                 interval          Tolerance,
    Count          Crop                          (days)              mg/kg
                                                                             

    South Africa   General                                         5.0

                   Table grapes                  28 - 42

                   Wine grapes,
                   peaches, apricots,
                   plums and prunes              14

    Spain          Pome fruit, grapes
                   strawberries,
                   onions, vegetables            20

    Sweden         General                        7

    Switzerland    Grapes                        21                1.0

                   Apples, pears                 21                0.5

                   Strawberries                  21                7.0

    United         Apples, blackberries,
    Kingdom        red and white currants,
                   black currants, grapes
                   grown outdoors,
                   loganberries, autumn-sown
                   salad onions, outdoor
                   lettuce, leaf brassicas,
                   seedlings under glass,
                   lettuce under glass,
                   treated with dust
                   formulation                   21

                   Strawberries, raspberries     14

                   Tomatoes under
                   glass, direct
                   spray on flowering
                   trusses                        3

    Yugoslavia     Strawberries                  14                2.0
                                                                   + 2.0 DMSA
                                                                             

    APPRAISAL

          The 1969 Joint Meeting, after reviewing the information on
    dichlofluanid, concluded that further work was necessary, before
    tolerances could be recommended. Some of the required information has
    been supplied.

          Data on the composition of technical dichlofluanid, including its
    impurities have been presented. Dimethyl-N-phenylsulphamide (DMSA) is
    known to be a metabolite, but detailed information on the pathway of
    dichlofluanid degradation, especially on the fate of the
    fluorine-containing moiety in plants, is not yet available. Studies
    are known to be in progress, and results will presumably be available
    to the 1976 Joint Meeting.

          The biological activity of dichlofluanid and similar fungicides
    (e.g. captan, folpet) is presumed to be localized in the
    trihalogeno-methylthio part of the molecule (Kühle et al., 1964). The
    inhibitory effect of dichlofluanid on yeast is antagonized by
    SH-compounds. The enzymes alcohol dehydrogenase and
    glycerinaldehyde-3-phosphate dehydrogenase and the co-factor co-enzyme
    A which contain essential SH-groups are strongly inhibited in vitro
    and/or in vivo (Lemperle et al., 1973). Dichlofluanid can therefore be
    regarded as an "SH-blocker."

          Processing of apples containing 0.8 mg/kg dichlofluanid plus 0.2
    mg/kg DMSA to stewed fruit, baby food or jelly resulted in residues of
    0.05 mg/kg dichlofluanid and 0.3 - 0.7 mg/kg DMSA. Cold pressed juice
    contained 0.2 mg/kg and heated juice 0.1 mg/kg dichlofluanid plus 0.7
    mg/kg DMSA.

          Wine originating from grapes with residues up to 48 mg/kg
    dichlofluanid contained up to 4 mg/kg DMSA and always less than 0.05
    mg/kg dichlofluanid. During crushing and fermentation much of the
    residue seems to be absorbed by the solid particles of the mash.

          In addition to the crops mentioned in FAO/WHO, 1970 the
    pre-harvest application of dichlofluanid has been extended to
    kohlrabi, leaf brassicas, melons, paprika, blackberries, loganberries,
    mulberries, raspberries, gooseberries, wheat and barley. In many cases
    dichlofluanid can be used as a substitute for other fungicides, e.g.
    dithiocarbamates and thiuram disulfides.

          Numerous data on rates and frequencies of application,
    pre-harvest intervals and the resulting residues of dichlofluanid and
    the degradation product DMSA have been supplied from Australia,
    Denmark, France, Germany, the Netherlands, Switzerland and the United
    Kingdom. The residue data, especially for currants, raspberries,
    strawberries and grapes, still show inconsistencies however. More data
    from supervised trials and on raw agricultural products moving in
    commerce are desirable for re-evaluating residue levels. Nevertheless
    the data on maximum residues so far obtained from supervised trials
    could be accommodated by the temporary tolerances recommended below,
    which seem to be in accordance with the toxicologically permissible
    level.

          The tolerances should refer only to the parent compound. This
    would substantially facilitate the determination of residues, because
    a gas-chromatographic multiresidue method could be used. This method
    is suitable for regulatory purposes in a range of crops, but further
    evaluation is desirable. DMSA can only be determined by a colorimetric
    method which is less suitable for regulatory purposes. The
    determination of DMSA is not considered necessary because available
    data indicate that excessive residues of this compound would be
    accompanied by excessive residues of dichlofluanid.

    RECOMMENDATIONS

          The following temporary tolerances are recommended. They refer to
    dichlofluanid only.

    TEMPORARY TOLERANCES

                                                       dichlofluanid
                                                          (mg/kg)

    Currants (red, black and white), grapes,
    raspberries                                              15

    Lettuce, strawberries                                    10

    Apples, pears, cucumbers, peaches                         5

    Beans (green, including pods), cherries,
    tomatoes                                                  2

    FURTHER WORK OR INFORMATION

    REQUIRED (by 1977)

    1.    Studies on absorption, distribution in various organs, and
          excretion of dichlofluanid in the rat.

    2.    Pharmacokinetics of the dichlorofluoromethylthio moiety.

    DESIRABLE

    1.    Metabolism studies on dichlofluanid.

    2.    Results from current studies on the pathway of degradation,
          especially the fate of the fluorine-containing moiety of the
          molecule, in and on plants. These data are expected to be
          available during 1975.

    3.    Further residue data from supervised trials, to resolve certain
          inconsistencies in the existing data or to provide new
          information, on blackberries, gooseberries, loganberries,
          mulberries, raspberries, currants, hops, kohlrabi, leaf
          brassicas, melons, onions, paprika and eventually wheat and
          barley.

    4.    Further residue data for raw agricultural products moving in
          commerce.

    5.    Further evaluation of the analytical method of Becker for
          regulatory purposes.

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    See Also:
       Toxicological Abbreviations
       Dichlofluanid (FAO/PL:1969/M/17/1)
       Dichlofluanid (Pesticide residues in food: 1977 evaluations)
       Dichlofluanid (Pesticide residues in food: 1979 evaluations)
       Dichlofluanid (Pesticide residues in food: 1981 evaluations)
       Dichlofluanid (Pesticide residues in food: 1982 evaluations)
       Dichlofluanid (Pesticide residues in food: 1983 evaluations)