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    INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY

    WORLD HEALTH ORGANIZATION



    TOXICOLOGICAL EVALUATION OF SOME
    FOOD COLOURS, EMULSIFIERS, STABILIZERS,
    ANTI-CAKING AGENTS AND CERTAIN
    OTHER SUBSTANCES



    FAO Nutrition Meetings Report Series 
    No. 46A WHO/FOOD ADD/70.36




    The content of this document is the result of the deliberations of the
    Joint FAO/WHO Expert Committee on Food Additives which met in Rome,
    27 May - 4 June 19691





    Food and Agriculture Organization of the United Nations

    World Health Organization



                   
    1 Thirteenth report of the Joint FAO/WHO Expert Committee on Food
    Additives, FAO Nutrition Meetings Report Series, in press;
    Wld Hlth Org. techn.  Rep. Ser., in press.


    SUCROSE ESTERS OF FATTY ACIDS

    Biological Data

    All substances tested contained dimethyl formamide.

    SUCROSE MONOPALMITATE

    Biochemical aspects

    The ester linkage is only hydrolysed with great difficulty by
    pancreatic or intestinal enzymes in vitro (Berry & Turner, 1960. The
    presence of sucrose monopalmitate had no effect on the absorption of
    Ca45 from aqueous CaCl2 solution when given intragastrically to six
    dogs. Estimation of blood activity up to six hours later showed only a
    delayed peak in the activity/time curve compared with plain aqueous
    CaCl2. Repetition in five dogs with induced impaired fat absorption
    secondary to ligation of the pancreatic duct showed higher Ca45
    absorption post-operatively from the control solution and test
    solution containing sucrose monopalmitate. Giving intramuscular 
    300 000 USP units Vit. D decreased the blood levels in both cases but
    less so for the solution containing sucrose monopalmitate (Tudisco,
    1961).

    Acute Toxicity

    Intragastic administration of 0.1 g to rats had no effect on the
    osmotic fragility of erythrocytes but intravenous administration to
    mice of 0.5 g/kg body weight produced considerable haemolysis (Tudisco
    R., 1965).

    Short-term studies

    Rat. In another experiment groups of ten male or female rats were
    given 0, 1 per cent., 2 per cent., 3 per cent., 5 per cent., 10 per
    cent. and 25 per cent. sucrose monopalmitate for 100 days. At the 2
    per cent. and 3 per cent. levels (female rats used only) there was
    marginal reduction in body weight gain of the test groups. At the 5
    per cent. level there were several deaths preceded by diarrhoea,
    weight gain was reduced but food efficiency high compared with
    controls.  The 10 per cent. and 25 per cent. groups did not survive
    the first week and had white semi-soft faeces. Histopathology showed
    nothing of note (Tudisco, 1961).

    Dog. Twelve male and 12 female young beagles were divided into four
    groups of three male and three female animals and fed a diet
    containing 0, 0.3 per cent., 1.0 per cent. and 3.0 per cent. sucrose
    monopalmitate for two years. Appearance, behaviour, appetite,
    elimination, body weight gains, urinalysis, organ weights,
    histopathology and organ function studies revealed no gross
    abnormalities or differences between test and control groups related
    to the compound (Hazelton Lab., 1966a).

    Long-term studies

    Rat. Groups of 25 male and 25 female albino rats were fed dietary
    levels of 0.3 per cent., 1 per cent. and 3 per cent. sucrose
    monopalmitate for two years. The control groups consisted of 50 males
    and 50 females. There was one death in the first six months, a male in
    the 3 per cent. test group, apparently caused by bilateral
    pyelonephritis.  Periodic records of body weight and food consumption
    disclosed a suggestion of lowered body weight gains in the 3 per cent.
    test groups. The first half-year, growth rate and the 78-week mean
    body weights of the 3 per cent. group females were significantly
    lowered; food consumption in the 3 per cent. group males was
    significantly lower at 26 weeks and slightly below that of controls in
    the first year; and the food efficiency of the 3 per cent. group
    females was slightly lowered in the first quarter year. No other
    differences were noted between the groups on the basis of observation
    of general appearance and behaviour, mortality and tumour incidence,
    haematology and urinalysis of pooled samples at 1, 3, 6, 12, 18 and 24
    months, and sacrifice with necropsy, organ weight measurement and
    histological examination of tissues on five males and five females
    from each group at three months and of all survivors at 24 months
    (Hazelton Lab. 1966b).

    A concurrent reproduction study over three generations was carried out
    on groups of eight male and 16 female rats over 22 months using
    sucrose monopalmitate at 0 or 1 per cent. of the diet. The parent
    generation (P) was kept for the whole study. Mean body weight gain,
    survival and food consumption of the P general showed no significant
    differences between controls and test group. The P generation was
    mated twice to give F1A and F1B filial generations. Of the F1A
    generation ten male and ten female pups were sacrificed for autopsy
    and the rest discarded. No significant findings were detected. The
    fertility index of the F1B test groups was significantly lower than
    that for the control groups due to adverse environment and unrelated
    to the test substance. Gestation, live birth and lactation indices
    were comparable between test and control groups. The F1B litter had
    slightly more frequent small weak pups with blue marks on the body but
    all survivors later developed normally. Sixteen F1B females and
    eight F1B males were mated twice to produce F2A and F2B filial,
    generations, all other F1B animals were discarded including the 24
    F1B animals used for mating.  The F2A and F2B generation
    developed normally both in control and the test group. Growth of the
    males in the test group was slightly higher and for the females
    slightly lower otherwise control and test group allowed no significant
    abnormalities. The F2A litter and all animals of the F2B litter
    except 16 females and eight males were discarded, the latter being
    mated twice to produce the F3A, and F3B filial generations. Five
    males and five females of these 24 F2B animals were sacrificed.
    Gross and histopathology of all major organs revealed no significant
    abnormalities. The F3A and F3B litters showed no difference from
    previous generations or between the test group and controls.

    Mean litter size, physical appearance and growth of litter were
    comparable among test and control groups for each generation and among
    the three filial generations.  The autopsies and histological
    examinations were normal (Hazelton Lab., 1965).

    SUCROSE MONOSTEARATE

    Biochemical aspects

    Surviving intestine has been shown capable of hydrolysing the
    glucosidic linkage of sucrose monostearate to glucose and fructose at
    one-quarter of the rate of sucrose hydrolysis. ▀-amylase and gluco- or
    fructo-invertase were unable to hydrolyse sucrose monostearate. Liver
    homogenate, but not intestinal mucosa homogenates, could oxidise
    sucrose monostearate. Only pancreatic juice was able to hydrolyse the
    ester linkage to a moderate degree (Berry & Turner, 1960).

    Short-term studies

    None available.

    Long-term studies

    None available.

    SUCROSE MONO OLEATE

    Biochemical aspects

    Rat liver homogenates hydrolysed the ester bond almost completely in
    60 minutes without attacking the glucosidic linkage, alpha-amylase,
    lipase and dog pancreatin are similarly effective. Invertases only
    released 3-6 per cent. of the existing glucosidic links. Intestinal
    mucosa and liver homogenates were able to oxidise 0.1 per cent. of the
    substrate (Berry & Turner, 1960).

    Acute toxicity

    No data available.

    Short-term studies

    Rat. Groups (unstated number) of rats received 0, 2 per cent., 5 per
    cent., 10 per cent. and 20 per cent. of ester in their diet for six
    months. Only at the 20 per cent. level (0.09 per cent.
    dimethylformamide) was there retardation of growth compared with
    controls. Soft faeces or mild diarrhoea occurred initially with 10 per
    cent. and 20 per cent. test diets but animals later became tolerant.
    On return to normal diet there was rapid regaining of full body weight
    (Oshima & Kajiwara, 1960).

    Long-term studies

    None available.

    SUCROSE MONOLINOLENATE

    Biochemical aspects

    Five rats with either their thoracic duct cannulated or having a bile
    and lymph fistula were given intragastrically an aqueous suspension of
    sucrose ester or linolenic acid. Linolenic acid increased in the lymph
    within 24 hours. No linolenic acid was found in the bile of animals
    given the ester during 48 hours and it appeared only slowly after 72
    hours in controls given linolenic acid. Rats with both bile and
    thoracic duct fistula showed a rise in the lymph content of linolenic
    acid within 24 hours and none in the bile even after 72 hours. Thus
    lymphatic absorption of the linolenic acid moiety was demonstrated.
    The percentage linolenic acid absorbed was the same whether the
    sucroester or the free acid was administered (Tudisco & Turner, 1963).

    Acute toxicity

    None available.

    Short-term studies

    None, available.

    Long-term studies

    None available.

    PALM OIL SUCROSE ESTERS

    Biochemical aspects

    When aqueous suspensions of Palm oil sucrose esters were given
    intragastrically to rats after pyloric ligation there was no evidence
    of digestive hydrolysis up to eight hours. In vitro use of
    intestinal juice on a substrate containing 1 per cent. palm oil
    sucrose esters for six hours produced only negligible hydrolysis of
    the ester and glucosidic linkage. A metabolic balance study on 35 male
    and female rats using food containing 5 per cent. palm oil sucrose
    esters and determining the amounts of ester given, the residue in
    stomach, gut, excreta and left over in food showed some 75 per cent.
    as unaccounted and, therefore, metabolised. Skeletal development as a
    measure of calcium absorption was determined by measuring growth of
    rat tails in two groups of ten rats receiving daily 1 ml of 50 per
    cent. palm oil sucrose esters suspension for ten weeks. Tail growth
    was more rapid in the test group during the first four weeks but later
    was not significantly different from controls (Balea et al., 1966).

    Acute toxicity

                                                                     

    Animal    Route         LD50            Reference
                        mg/kg body weight
                                                                     

    Rat       oral        > 30 000          Balea, 1963
                                                                     

    Mice receiving intravenous doses of 1-2 g/kg body weight showed no
    haemolysis (Tudisco, 1965).

    Short-term studies

    Rat. Groups of eight rats received 5 per cent. or 10 per cent. palm
    oil sucrose ester. in their diet for 150 days without showing any
    deleterious effect on body weight gain (Tudisco & Chiancone, 1965).
    Three groups of ten rats were fed diets containing 10 g/kg body weight
    lard with either 0, 50 ml/kg body weight 10 per cent. palm oil sucrose
    esters and 10 ml/kg body weight 50 per cent. palm oil sucrose esters
    for six weeks. No gastro-intestinal disturbances were seen. Test
    groups showed a slightly greater body weight gain and slightly larger
    fat deposits in their carcass compared with controls (Balea et al.,
    1966). When the compound was fed to groups of eight male and eight
    female rats at 0, 3 per cent., 5 per cent. and 10 per cent. levels of
    their diet for 100 days no difference was noted between tests and
    controls regarding general condition. Test groups showed a slightly
    greater weight increase. Haematology, organ weights and gross autopsy
    were comparable in tests and controls, while histopathology revealed
    no abnormalities in the test group (Balea, 1963). In a similar
    experiment with 8-10 female rats per group, these received 0, 5 per
    cent. and 10 per cent. the compound in their diet for 100 days. No
    abnormal ties related to the test substance were detected in general
    appearance, food consumption, body weight gain, food efficiency,
    haematology, gross autopsy findings, histology (thyroid, liver,
    adrenal) and biochemistry (plasma and liver cholesterol,
    phospholipids, total lipids) only the liver lipids of test animals
    were significantly raised (Tudisco, 1963a).

    Two groups of ten rats received 0 or 2 per cent. palm oil sucrose
    esters in their diet for three-and-one-half months. No deleterious
    effect was noted on mating, reproduction, litter number and litter
    size. Pups gained weight satisfactorily during weaning with better
    performance in the test group. Lactation was unaffected and no
    external foetal abnormalities were seen (Fernando, 1964).

    Long-term studies

    Rat. Groups of 15 male or female rats were fed diets containing 0 or
    0.5 per cent. of the compound for 14 months. No tumours were seen at
    the end of 13 months nor any growth abnormalities in male rats, Female
    rats showed a temporary lowering of growth rate during the eighth to
    tenth week. Three animals were examined histologically and no evidence
    of abnormality attributable to palm oil sucrose esters was found.
    Haematologic findings and blood proteins were comparable in test and
    control groups (Chiancone at el., 1963).

    Groups of 12 male rats were fed 0 or 10 per cent. of the ester as only
    source of lipids in their diet for 25-28 months. No abnormalities were
    found with respect to body weight, general health, mortality,
    haematology, plasma proteins and food efficiency. Tumour incidence was
    similar in test groups and controls. Visceral abnormalities were
    equally distributed (Tudisco & Chiancone, 1965).

    A two-generation study was carried out on 15 males and 15 female rats
    given 0 and 5 g/kg body weight palm oil sucrose esters containing 68
    ppm of dimethyl formamide daily. The parent generation (P) was
    observed for two years. Body weight gain, external appearance and
    incidence of tumours were identical to controls. Three male animals
    died from intercurrent respiratory disease, three female animals
    either died or were sacrificed, the remaining 12 animals survived 24
    months. No haematological abnormalities were detected and blood
    proteins had a normal electrophoretic pattern. At autopsy no
    significant lesions ware found in the test group which were not also
    found in the controls. Histology was similarly comparable for tests
    and controls. Five male and five female rats (P generation) were mated
    after nine months' treatment to produce 23 F1 generation animals.
    The F1 generation was observed for 21 Months. Body weight gain was
    similar to controls and no spontaneous deaths or illnesses occurred.
    Five male and five female survivors were autopsied after 21 months. No
    evidence of malignant tumours or other significant abnormalities was
    found which differed from the controls. Five male and five female F1
    animals were mated after nine months to produce 30 F2 generation
    animals. The F2 generation was observed for 14 months. All litter
    mates gained weight identical to controls. Five male and five female
    F2 animals were autopsied after 14 months. No malignant tumours or
    other pathological conditions were found which did not also occur in
    the controls. No other significant abnormalities were seen. Seven from
    50 animals examined showed hepatic hyper-function. No evidence of any
    adverse effects on fertility and foetal development was seen
    (Mosinger, 1964b).

    LARD AND TALLOW SUCROSE ESTERS

    Biochemical aspects

    Each of four dogs aged 5-6 years were given intragastrically I131
    -labelled triolein in olive oil, oil/water emulsion and oil/water
    emulsion containing lard sucrose esters and the plasma activity

    determined as a measure of absorption. Each animal served as its own
    control, the administrations being separated by seven-day intervals.
    With olive oil peak absorption occurred after six hours with a fall
    between six and eight hours. The oil/water emulsion gave inconsistent
    patterns while oil/water emulsion with lard sucrose esters had a peak
    absorption of four hours and less abrupt fall during 4-8 hours. Lard
    Sucrose esters did not appear to interfere with intestinal fat
    absorption (Tudisco, 1961).

    Calcium absorption using Ca45Cl2 was tested in eight dogs using
    aqueous CaCl2 solution. CaCl2 in oil/water emulsion and CACl2 in
    oil/water emulsion with lard sucrose ester. All doses were
    administered intragastrically and blood activity determined
    subsequently. Each dog acted as its own control. The presence of lard
    sucrose ester had no consistent effect on CA45 absorption (Tudisco,
    1961).

    Each of four dogs aged one year received simultaneous I131-labelled
    triolein and Ca45Cl2 either in oil/water emulsion and oil/water
    emulsion with lard sucrose esters. All doses were given
    intragastrically. Lard sucrose esters bad no effect on the pattern of
    absorption. Experiments were repeated in young and old dogs with
    ligated pancreatic duct and administering intragastrically
    I131-labelled triolein in oil/water emulsions or oil/water emulsions
    with lard sucrose ester. Pre- and post-operative blood samples showed
    that no significant effect was exercised by the ester (Tudisco, 1961).

    As much as 100 g fat as lard sucrose ester may be administered to dogs
    and human subjects without producing plasma turbidity or an increase
    in excreted faecal fat (Berry & Turner, 1960). Humans have been
    maintained for short periods on this material as the sole source of
    dietary lipid (Tudisco, 1965).

    Rats tolerated doses of 5 g/kg body weight lard sucrose ester every
    two hours up to a total of 15 g; 10 g/kg body weight every two hours
    up to a total of 30 g produced diarrhoea and death from
    overdistension. Single doses of 3.8 g/kg body weight in dogs had no
    effect. Rabbits tolerated 2.0 g/kg body weight single doses without
    deleterious effects (Tudisco & Chiancone, 1965). Intragastric
    administration of 1 g to rats or 20 per cent. of the ester in their
    diet produced no osmotic fragility of erythrocytes. Rabbits given
    3-6 g intragastrically also showed no osmotic fragility rise.
    Intravenous administration of 1-2 g/kg body weight to mice had no
    haemolytic effect (Tudisco, I965).

    Single oral doses of lard sucrose ester (48 per cent. sucrose ester,
    12 per cent. triglycerides, 21 per cent. diglycerides, 19 per cent.
    monoglycerides) were administered to four adult dogs aged 4-6 years as
    follows: 56 g lard sucrose ester. after a long interval 40 g refined
    lard, after a long interval 56 g lard sucrose ester and 40 g refined
    lard. Thus each animal served as its own control. No gastro-intestinal
    disturbances, toxic symptoms or abnormal findings were discovered when

    blood sugar, total lipid, glyceride, cholesterol, phospholipid and
    percentage fatty acid composition were estimated. Plasma turbidity was
    lowest in the dog receiving the ester alone (Tudisco, 1963b).

    Short-term studies

    Rat. Groups of 12 animals received 0, 5 per cent., 10 per cent. and
    25 per cent, lard sucrose ester daily in their diet for 15 days
    without adverse effects except that the test group at the 25 per cent.
    level showed reduced body weight gain (Tudisco & Chiancone, 1965).

    In another experiment groups of 12 rats each received for 200 days 5
    per cent., 10 per cent. and 25 per cent. lard sucrose ester in their
    diet while controls received 3.6 per cent., 7.7 per cent. and 18 per
    cent. lard. Body weight gain was reduced at the 10 per cent. and
    especially the 25 per cent. levels. Food consumption and food
    efficiency were reduced at both the 10 per cent. and 25 per cent.
    level. No abnormal findings were seen in haematology, and clinical
    tests (blood glucose, plasma lipids, plasma proteins, liver lipids).
    There were no undue gastro-intestinal symptoms. Only test animals at
    the 25 per cent. level were autopsied; there were no significant
    abnormalities in organ weights or histology of kidney, spleen, thyroid
    and adrenals. The liver showed frequent "steatosis" (Lepetit Lab.,
    (1961)). When male rats were given a diet containing 25 per cent. lard
    sucrose ester or 18 per cent. lard and 7 per cent. sucrose for 200
    days there was no adverse effect noticeable in body weight gain, food
    consumption, survival. Clinical findings which could not be ascribed
    to high fat diet were absent. Only the total liver fat of tests and
    controls was raised while the blood cholesterol of controls was also
    higher. Autopsy revealed no significant organ changes (Tudisco, 1967).

    Long-term studies

    Rat. Groups of 9-11 male rats were fed 3.6 per cent. lard, 5 per
    cent. and 10 per cent. lard sucrose ester in their diet as only source
    of lipids for 24-28 months. No abnormalities were found with respect
    to body weight, general health, mortality or haematology. Tumour
    incidence was similar in test groups and controls. Visceral
    abnormalities were equally distributed (Chiancone, et al., 1963,
    Tudisco & Chiancone, 1965).

    A two-generation study was carried out starting with 15 male and 15
    female rats receiving 0 and 5 g/kg body weight lard sucrose ester
    containing 52 ppm of dimethyl formamide for two years. The parent
    generation was observed for two years. Four females died or were
    sacrificed for reasons unrelated to the test substance while eleven
    animals survived. Three males died from respiratory disease. After 24
    months autopsies showed no malignant tumours or other conditions to be
    present which did not also occur to a similar extent in controls.
    Haematological findings were normal and electrophoresis or plasma
    proteins showed nothing unusual. Five males and five females were
    mated after nine months to produce 35 F1 generation pups. All F1,
    animals gained weight satisfactorily without spontaneous deaths and

    were kept for 21 months. Five males and five females were examined
    after 21 months without showing any malignant or other tumours not
    also present equally in the controls. No abnormal findings were seen.
    Five males and five females of the F1 generation were mated after
    nine months to produce 32 F2 generation animals. The F2 generation
    survived for 14 months without showing any abnormalities as regards
    body weight gain or mortality. Five male and five female F1 animals
    were autopsied at 14 months without showing any unusual tumours or
    benign lesion or other condition not occurring equally frequently in
    the controls. No adverse effects were seen on fertility or foetal
    development (Mosinger, 1964a).

    Groups of 15 male or female rats were fed diets containing 0 or 0.5
    per cent. tallow sucrose ester for 14 months. No tumours were seen at
    the end of 13 months nor any growth abnormalities in male rats. Female
    rats showed a temporary lowering of growth rate during the eighth to
    tenth week. Three animals were examined histopathologically and no
    evidence of abnormality attributable to the ester was found.
    Haematology and blood protein investigation showed comparable results
    in tests and controls. No animal died. (Oshima & Kajiwara 1960)

    SUPPLEMENTARY INFORMATION ON DIMETHYL FORMAMIDE

    Biological Data

    Biochemical aspects

    Dimethyl Formamide is an industrial solvent. Following intravenous
    injection of 6 g into cats some 9-16 mg appear in the urine, 3-5
    mg/100 ml in the blood, 6-9 mg in the stomach and 0.04-1 per cent. in
    the expired air over two hours. Rabbits excrete 10-15 per cent. of the
    dose in their urine (Massmann, 1956).

    Pharmacological action in animals is similar in several species.
    Intraperitoneal injection causes a hyperglycaemic phase of 24 hours
    (Massmann, 1956). Intravenous injection in rats and cats produces
    shortlived hypotension followed by brief hypertension. Dogs show very
    small effects. Dogs respond by strong duodenal contractions and
    faster, shallower, breathing. Bile flow in dogs and diuresis in rats
    were not affected (Auclaire & Hameau, 1964),

    Acute toxicity
                                                                                             

    Animal       Route                     LD50                Reference
                                      mg/kg body weight
                                                                                         

    mouse        oral                 5900, 3750               Kutzsche, 1965;
                                      6580                     Stasenteva, 1961;
                                                               Auclair & Hameau, 1964
    "            intraperitoneal      1722, 2870, 3900         Davis & Jenner, 1959;
                                      4324                     Massmann,
                                                               Kutzsche, 1965;
                                                               Auclair & Hameau, 1964
    "            intravenous          2500                     Kutzsche, I965;
                                                               Auclair & Hameau, 1964
    rat          oral                 3480, 4000, 7000         Tudisco, 1965;
                                                               Massmann, 1956;
                                                               Stasorkova, 1961;
                                                               Smyth & Carpenter, 1948
    "            subcutaneous         3570, 3500               Massmann, 1956;
                                                               Stasenkova, 1961
    "            intraperitoneal      3800, 1260               Heath, 1962;
                                                               Massmann, 1956
    guinea-pig   intravenous          1030                     Kutzsche, 1965
    rabbit       intraperitoneal      approx. 1000             Massmann, 1956
    "            intravenous          1800                     Kutzsche, 1965
    cat          intraporitoneal      400                      Massmann, 1956
    dog          intravenous          470                      Kutzsche, 1965
                                                                                         
    
    Many animals especially cats and rabbits, showed consistently severe
    fatty degeneration and necrosis of the liver with associated weight
    loss and anorexia (Martelli, 1960; Massmann, 1956). On the other hand,
    groups of ten rats received 0.28 mg/kg body weight daily subcutaneous
    for one month and 0.47 mg/kg body weight daily orally. There were no
    differences from controls as regards growth, haematological indices or
    gross autopsy (Auclaire & Hameau, 1964). Oral administration of 1.84
    mg/kg daily for 10 days caused the death of 75 per cent. of animals
    with evidence of gastric and pulmonary haemorrhage (Auclaire & Hameau,
    1964).

    Single oral, intraperitoneal, subcutaneous, intramuscular and intra
    venous doses of 5.3 g/kg or single applications of 3.5 g/kg caused
    liver necroses in some mice but intravenous doses of one and 2.5 g/kg
    in rats had no effect (Kutzsche, 1965).

    Single toxic doses in mice produced depression and hind leg paralysis.
    In dilute form (20 per cent. in water) dimethyl formamide had about
    one quarter the toxicity of the undiluted material in mice (Davis &
    Jenner, 1959). Hofmann (1960) observed no effect on the liver by the
    bromsulphthalein test or histolologic examination in rats, rabbits and
    cats following intraperitoneal injection of 1.5 ml/kg.

    Dimethyl formamide is absorbed through the skin in man and in rabbits
    with some animal deaths in rats occurring from exposure to 75 per
    cent. dimethyl formamide (Massmann, 1956). Rabbits died with liver
    damage after skin application (Kutzsche, 1965). Dimethyl formamide
    irritates human and rabbit skin and rabbit conjunctiva (Kutzsche,
    1965). Inhalation studies on rat, mouse, dog, guinea-pig and rabbit
    revealed variable effects (Clayton et al., 1963; Hofmann, 1960) but
    other experiments in rats and cats showed cumulative effects such as
    liver necroses and haemorrhagic enlargement of the kidneys in rats
    with fatty degeneration of the liver in cats over a range of 100-450
    ppm in air (Massmann, 1956). In chick embryo tests it has a
    comparatively low toxicity (McLaughlin et al., 1964).

    Special studies

    No teratogenic or cytotoxic activity was shown in mice after oral,
    intraperitoneal or percutaneous application (Oettel & Frohberg, 1964).
    Dimethyl formamide increases biological activity as a synergist, acts
    bacteriostatically and as antitrichomonas (Kutzche, 1965).

    Short-term studies

    Mouse. When mice were fed 50 mg dimethyl formamide/kg body weight
    for 2.5-4 months no adverse effects were noted (Zamyslova and
    Smirnova, 1960). Ten mice received 0.55 g/kg of a 30 per cent.
    solution six days a week for six weeks, Two animals died, all
    survivors had liver necroses and fatty infiltration (Kutzsche, 1965).

    Rat. Three groups of ten male rats were given 0, 0.003 per cent.,
    and 0.03 per cent. dimethyl formamide in their diet for 90 days.
    Growth was comparable with controls: at 0.03 per cent. slight
    congestion of kidneys was noted. At 0.003 per cent. and 0.03 per cent.
    a few cases of mild hepatic steatosis were seen (Tudisco, 1965).

    In another study rats were given 0.02 per cent., 0.1 per cent. and 0.5
    per cent. dimethyl formamide in their diet for 90 days. At the 0.1 per
    cent. level and higher there was weight loss, reduced food intake and
    liver enlargement. At the 0.5 per cent. level anaemia, leucocytosis,
    liver lesions and hypercholesterolaemia were seen (Sherman and Read,
    1944).

    The oral administration to rats of 1.88 g/kg of the undiluted dimethyl
    formamide daily for 10 days killed 75 per cent. of the animals with
    marked gastric and pulmonary haemorrhage. In groups of 10 rats
    receiving 0.28 g/kg subcutaneous or 0.47 g/kg orally daily for
    1 month, there were no differences from controls in growth,
    haematological indices or gross autopsy findings (Auclaire & Hameau,
    1964).

    Rabbit. Of five animals receiving 45 g/kg orally for four weeks, two
    had liver necroses yet normal liver function tests (Kutzsche, 1965).

    Inhalation of 300 ppm for 6 hours a day for 10 months caused no
    clinically demonstrable liver or kidney damage (Hofmann, 1960).

    Cat. Inhalation of 300 ppm for 6 hours a day for 10 months caused no
    clinically demonstrable liver or kidney damage. Inhalation of 0.1 vol.
    per cent. for 10 weeks also produced no clinical or histological
    evidence of liver injury (Hofmann, 1960).

    Dog. Two dogs received orally 25 mg/kg five times per week for ten
    weeks and then 50 mg/kg body weight for two weeks. Only BP changes
    were seen but no other pathological changes (Kutzsche, I965).

    Long-term studies

    None available.

    Comments on Dimethyl Formamide

    The metabolism of this compound is not known. The no-effect level
    obtained in 90-day tests in rats is 0.02 per cent. At highest levels
    it produces carbons liver lesions apparently similar to those caused
    by certain chlorinated hydrocarbons or nitrosamines. Adequate
    long-term studies in several species are required because of its
    cumulative activity with particular emphasis on the nature of the
    liver pathology.

    Comments on Sucrose Esters of Fatty Acids

    It seems reasonable, in evaluating these compounds, to consider mainly
    their metabolic fate provided that it is similar to their metabolic
    behaviour in man. However, the available metabolic studies are not
    adequate to reveal the pathways of individual products. The long-term
    feeding studies in rats on some representative compounds are adequate.
    A two-year study has been reported on sucrose monopalmitate. The
    presence of dimethyl formamide at levels up to 50 ppm has not
    introduced an adverse effect in the long-term toxicity tests on
    sucrose esters. However, most of the short-term and all long-term
    tests have been performed in the rat only.

    EVALUATION

    Level causing no toxicological effect in the rat

    One per cent. (= 10 000 ppm) in the diet equivalent to 500 mg/kg body
    weight/day.

    Estimate of acceptable daily intake for man

                                            mg/kg body weight

    Temporary acceptance                         0-2.51

    Further work required by June 1974

    Metabolic studies on representative individual sucrose esters.
    Two-year studies on another sucrose ester, in a non-rodent mammalian
    species.

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    Berry, J. F. & Turner, A. D. (1960) J. Amer. Oil Chem. Soc., 37, 302

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    Hazelton Laboratories (1966a) Report to Sucro-Chemical Division dated
    25 March 1966


                   

    1 This applies to material the dimethyl formamide content of which
    does not exceed 50 ppm.

    Hazelton Laboratories (1966b) Report to Sucro-Chemical Division dated
    20 April 1966

    Heath, D. F. (1962) Biochem. J., 85, 72

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    See Also:
       Toxicological Abbreviations
       SUCROSE ESTERS OF FATTY ACIDS (JECFA Evaluation)