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. ß-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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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
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