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Toxicological information

Carcinogenicity

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Description of key information

Studies conducted with glycerides, fatty acids, tocopherols and squalene, indicated that these substances are not carcinogenic. Moreover, predictions for ‘Oils, vegetable, deodorizer distillates’ (based on the main constituents) from the Danish QSAR database and Toxtree (v.1.6) model did not reveal any structural alerts for carcinogenicity. Most of these substances have a long history of safe use in nutritional, cosmetic and/or industrial applications. Some of the constituents (e.g. certain fatty acids and tocopherols) are listed as GRAS and/or demonstrate antitumourigenic activity under certain conditions (e.g. tocopherols and squalene; see Section 5.8 of the CSR).

Key value for chemical safety assessment

Carcinogenicity: via oral route

Link to relevant study records

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Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
A study was conducted to investigate the carcinogenic effects of the constituent sodium oleate following administration of 2.5% and 5% doses in drinking water to a group of 50 male and female F344 rats for 108 weeks.
GLP compliance:
not specified
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Co. Japan Inc. Atsugi
- Age at study initiation: 7 weeks
- Housing: In wire cages ( 2 males or females/cage)
- Diet (e.g. ad libitum): Yes, basal CRF-1 diet
- Water (e.g. ad libitum): Yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1°C
- Humidity (%): 55 ± 5%
Route of administration:
oral: drinking water
Vehicle:
other: Distilled water
Details on exposure:
Groups of 50 male and 50 female F344 rats, initially 7 wk old, were administered 'fatty acids and fatty acid salts' (sodium oleate) for 108 wk at concentrations of 2.5 and 5.0% in the drinking-water. Control rats were given distilled water only. Both diet and distilled water, with or without oleate, were available ad libitum.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
None
Duration of treatment / exposure:
108 wks
Frequency of treatment:
Daily
Post exposure period:
No
Remarks:
Doses / Concentrations:
2.5 and 5%
Basis:
nominal in water
No. of animals per sex per dose:
50 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels for the test were chosen on the basis of a preliminary subchronic (13-wk) study with six dose levels in the 0-10% range.
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: every 2 or 4 wk

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: twice weekly

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At week 108
- How many animals: 10 animals each/sex/dose group

CLINICAL CHEMISTRY: Yes

URINALYSIS: Yes
- Time schedule for collection of urine: At week 108

Sacrifice and pathology:
At autopsy included examination of tumors and other leisons in the brain, spinal cord, salivary, pituitary and thyroid (including parathyroid) glands, thymus, lungs including the trachea), heart, oral cavity and tongue, oesophagus, stomach, duodenum, jejunum, ileum, caecum, colon, rectum, liver, pancreas, spleen, adrenal glands, lymph nodes, skin, subcutis, preputial/clitoral gland, ear duct, eyes and Harderian glands, musculature, sternum, femur and nasal cavity.

Turnout masses, and all organs and tissues were fixed in buffered neutral 10% formalin and sectioned. The sections were routinely stained with haematoxylin and eosin, other specific stains being used when necessary.
Other examinations:
None
Statistics:
Student's t test used for haemotology and serum biochemistry data analysis
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
effects observed, treatment-related
Details on results:
CLINICAL SIGNS AND MORTALITY: Numbers of surviving animals at week 75 (and at week 108 in parenthesis) were:
For male 49 (42) in the 5% group, 49 (37) in the 2.5% group and 48 (30) in the control group, and for females, 47 (38) and 50 (36) in the 5 and 2.5% groups, respectively, and 48 (29) in the control group.

WATER CONSUMPTION: Sodium oleate slightly depressed water consumption in the females, but not in the males

CLINICAL CHEMISTRY:
- A slight deacrease in serum bilirubin was the only statistically significant finding in the serum analyses in the males
- A decrease in lactic dehydrogenase activity and an increase in blood urea nitrogen seen in the females receiving 2.5% oleate was not apparent in the higher dosage group.
- No significant differences between treated and control rats of either sex in the values for glutamic-oxalacetic and glutamic-pyruvic transaminases, alkaline phosphatase, zinc sulphate turbidity test, thymol turbidity test, 7-glutamyl transpeptidase, triglycerides, cholesterol, total proteins, albumin/globulin ratio or creatinine

BODY WEIGHT AND WEIGHT GAIN: No significant difference between control and treated animals in the growth curve. Sodium oleate slightly reduced the body-weight gain in the males at the highest dose, but not in females.

HAEMATOLOGY: No significant differences between the controls and treated rats in the numbers of erythrocytes or leucocytes, in the haemoglobin content or in the haematocrit readings

URINALYSIS: No significant differences between the controls and treated rats; although urinary protein was detected in two of ten males given 2.5% oleate, and ketones were demonstrated in the urine from two of ten females given 5% oleate and from one of the ten male control rats.
- No urine samples gave positive results for glucose or urobilinogen and the pH was 6 or 7 in all treated and control rats

ORGAN WEIGHTS: At 5% oleate, the mean weights of the liver of males and of the heart, pancreas and adrenals of females were significantly lower than those of the respective controls, while the weight of the thymus in the females was higher.

HISTOPATHOLOGY: NEOPLASTIC (if applicable): For the males, tumors were found in 96% of the 5%-oleate group, in 100% of the 2.5% group and in 98% of the controls, while the corresponding figures for the females were 56, 72 and 56%. These high proportions of tumor-bearing rats were due to the high incidences of tumors of the testis, thyroid, pituitary gland, pancreas and uterus. The incidences of turnouts of the pituitary gland (in males), thyroid gland (in females), adrenal gland (in males), pancreas (in males) and uterus were slightly higher in treated rats than in the controls, but the differences were not statistically significant except in the case of the pancreatic tumors.
Key result
Dose descriptor:
NOAEL
Effect level:
5 other: % in diet
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no treatment-related effects observed on recorded parameters
Key result
Critical effects observed:
no

None

Conclusions:
Under the test conditions, 2.5 and 5 % of the 'fatty acids and fatty acid salts' (sodium oleate) administered in drinking water to F344-rats of either sex, for 108-weeks, have no carcinogenic potential.
Executive summary:

A study was conducted to investigate whether the constituent fatty acids and fatty acid salts' (sodium oleate), in drinking water, had carcinogenic impact on the F344 rats. Groups of 50 male and female rats were given sodium oleate for 108 -wks at the concentration of 2.5 and 5% in drinking water. Parameters monitered were survival, body weights, sodium oleate and water consumption, clinical chemistry, blood and urine analysis, terminal body weights, organ weights and tumour incidences. Under the conditions of the study, 'fatty acids and fatty acid salts' (sodium oleate) did not show any carcinogenic potential in F344-rats (Hiasa, 1984).

Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
A study was conducted to investigate whether the constituent palm oil, as a dietary fat, has an impact on mammary carcinogenesis in female rats induced by DMBA. Groups of 20 female rats were given one single dose DMBA and after three days were fed semisynthetic diets containing 20% soybean oil, corn oil, crude palm oil, deodorized palm oil or metabisulfite-treated palm oil, respectively, for a duration of 5 months. At autopsy, blood was collected from the tumor-bearing rats. The tumors were examined and lipid extractions were made for analysis of fatty acid profile, as well as tocopherols, tocotrienols and carotenes content.
GLP compliance:
not specified
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
The rats were obtained from the Animal Breeding Unit, University of Malaya at 45 d of age. They were allocated to groups of 20 each, put on laboratory chow diet and housed in an experimental room with 12 h of light and 12 h of darkness. Diets were formulated according to Newberne. Water and food were given ad libitum.
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
At 50 d of age, all rats were given a single dose of 5 mg of DMBA in 0.25 mL of CO intragastrically. All rats were continued on laboratory chow diet for another 3 d after DMBA administration. Thereafter, they were fed semisynthetic diets containing 20% by weight of soybean oil, corn oil, deodorized palm oil, palm oil and metabisulfite-treated palm oil.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
None
Duration of treatment / exposure:
6 months
Frequency of treatment:
Daily
Post exposure period:
No
Remarks:
Doses / Concentrations:
20% by weight
Basis:
nominal in diet
No. of animals per sex per dose:
20 animals per sex per dose
Control animals:
no
Details on study design:
No data
Positive control:
No
Observations and examinations performed and frequency:
The rats were weighed weekly. The appearance of mammary tumors was recorded. Before autopsy, blood was taken from ether-anesthetized tumor-bearing rats.
Sacrifice and pathology:
At autopsy, the presence of mammary tumors in each rat was carefully examined and suspected tissues were preserved in 10% formalin. Solid mammary tumors were excised, frozen and kept at -20°C.
Other examinations:
Lipid Extraction.
Fatty Acid Analysis
Analyses of tocopherols, tocotrienols and carotenes
Statistics:
No data
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Haematological findings:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: neoplastic:
effects observed, treatment-related
Details on results:
During the course of the experiment, rats fed different dietary fats had similar rate of growth. Rats fed 20% corn oil or soybean oil have higher tumor incidence than rats fed on palm oil diets. However, differences of mean tumor latency periods among the groups were not statistically significant. At autopsy, rats fed on high corn oil or soybean oil diets had significantly more tumors than rats fed on the three palm oil diets.
Relevance of carcinogenic effects / potential:
Under the conditions of the study, the test substance had a non-promoting effect on chemically-induced mammary carcinogeneis in rat.
Key result
Dose descriptor:
NOAEL
Effect level:
20 other: % in diet (i.e. ca. 10,000 mg/kg bw/day)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: non-promoting effect on chemically induced mammary carcinogenesis in female rats
Key result
Critical effects observed:
no

None

Conclusions:
Under the test conditions, diet containing high amount of the substance do not promote chemically-induced mammary tumorigenesis in female rats.
Executive summary:

A study was conducted to investigate whether the constituent ‘glycerides, C16 -18 and C18 -unsatd.’ (as palm oil), as a dietary fat, had an impact on mammary carcinogenesis in female rats induced by DMBA. Groups of 20 female rats were given one single dose DMBA and after three days were fed semisynthetic diets containing 20% of the substance, soybean oil, corn oil, deodorized palm oil or metabisulfite-treated palm oil, respectively, for a duration of 5 months. At autopsy, blood was collected from the tumor-bearing rats. The tumors were examined and lipid extractions were made for analysis of fatty acid profile, as well as tocopherols, tocotrienols and carotenes content. Under the conditions of the study, the substance was shown to have a non-promoting effect on chemically induced mammary carcinogenesis in female rats (Sundram, 1989).

Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
21 d old Sprague Dawley rats were fed ad libitum on diets containing 5% (normal fat) corn oil, 20% (high fat) corn oil, 20% palm oil, 20% beef tallow or 20% lard. These diets were formulated so that all nutrients except fat and dextrose were equivalent on a caloric basis. Rats in the various dietary treatments (32/group) were orally gavaged 7.5 mg 12-dimethylbenz(a)anthracene (DMBA) at 52 d of age. One week later, the rats were shifted to a 5% corn oil control diet for the rest of the experiment. Animals were placed on a normal diet to determine the early effects of the various high fat diets on subsequent mammary tumourigenesis.
GLP compliance:
not specified
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
21 d old rats were purchased from Charles River Breeding Laboratories (Wimington, MA). They were immediately divided into different treatment groups and were allowed to feed ad libitum on their diet respectively. The diets were formulated according to the method of Newberne P. M. et al. 1978 (Control of diets in laboratory animal experimentation. Inst. Lab. Anim. Resour. News, 21: A1-A12) so that all nutrients except fat and dextrose were equivalent on caloric basis. All animals were housed in suspended metal cages in a temperature-regulated room (22 ± 0.5°C) and light controlled (12 h light, 12 h dark in experiment 1, and 14 h light, 10 h dark in experiment 2), and were allowed free access to drinking water.
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
Group 1: 5% (normal fat) corn oil
Group 2: 20% (high fat) corn oil
Group 3: 20% palm oil
Groupv4: 20% beef tallow
Group 5: 20% lard

Experiment 1: The various treatment groups were given p.o. 7.5 mg DMBA at 52 days of age. One week following DMBA administration all rats were switched to the 5% corn oil control diet for the remainder of the experiment.

Experiment 2: 21 day old rats were fed the various diets in the same manner as in experiment 1.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
None
Duration of treatment / exposure:
Approximately 6 months
Frequency of treatment:
Daily
Post exposure period:
No
Remarks:
Doses / Concentrations:
20%
Basis:
nominal in diet
No. of animals per sex per dose:
32
Control animals:
yes
Details on study design:
According to the author, high dietary fat intake significantly enhances mammary tumorigenesis in rats. The mechanism(s) by which high fat diet bring about this stimulation is not fully understood. However, evidence suggest that dietary fat influences tumor growth during the promotional phase of mammary carcinogenesis. The purpose of this investigation was to determine whether diets high in animal or vegetable fat affect mammary tumorigenesis when fed to rats only prior to and during the initiation phase of carcinogenesis.

Dietary treatments were selected as representative saturated and/or unsaturated animal and vegetable fats, thereby providing the opportunity to examine their comparative effects on mammary tumorigenesis and endocrine functions in rats.
Positive control:
No
Observations and examinations performed and frequency:
Tumor measurements and body weight were recorded weekly.
Sacrifice and pathology:
15 - 30% of the animals in each treatment group had to be sacrificed before termination of the experiment, when they became moribund from increased tumor burden.

19 weeks after DMBA administration, all remaining rats were killed.
Other examinations:
No
Statistics:
No data
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Haematological findings:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: neoplastic:
effects observed, treatment-related
Details on results:
Mammary tumor development in rats fed 20% corn oil or palm oil diets was similar to that of controls (5% corn oil).
Key result
Dose descriptor:
NOAEL
Effect level:
20 other: % in diet (i.e. ca. 10,000 mg/kg bw/day)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: non-promoting effect on chemically induced mammary carcinogenesis in female rats
Key result
Critical effects observed:
no

None

Conclusions:
Under the study conditions, diets high in vegetable fat containing either the saturated substance or polyunsaturated (corn oil) fatty acids have no effects on subsequent mammary tumorigenesis.
Executive summary:

A study was conducted to determine the effects of diets high in the constituent ‘glycerides, C16 -18 and C18 -unsatd.’ (as 20% palm oil or vegetable fat) on mammary tomourigenesis when fed to rats prior to or during the initial phase of carcinogenesis. 21 d old Sprague Dawley rats were fed ad libitum on diets containing 20% of the substance, 5% (normal fat) corn oil, 20% (high fat) corn oil, 20% beef tallow or 20% lard. Rats in the various dietary treatments (32/group) orally gavaged 7.5 mg 12 -dimethylbenz(a)anthracene (DMBA) at 52 d of age. One week later, they were shifted to a 5% corn oil control diet for the rest of the experiment. Animals were placed on a normal diet to determine the early effects of the various high fat diets on subsequent mammary tumourigenesis. Under the study conditions, that diets high in vegetable fat containing either the saturated substance or polyunsaturated (corn oil) fatty acids have no effects on subsequent mammary tumorigenesis (Sylvester, 1986).

Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
The effects of dietary administration coconut oil at a dose equivalent to 54% of calories in the diet was investigated in male and female Wistar rats in a chronic toxicity study (104 wks).
GLP compliance:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Central Breeding Station TNO-Zeist
- Age at study initiation: Newly weaned SPF-Wistar rats
- Weight at study initiation: 41.7 g
- Diet (e.g. ad libitum): Ad libitum

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
- Mixing appropriate amounts with (Type of food): Normal diet containing 6 calorie (cal)% soybean oil as essential fat source and 54 cal% of experimental fat (i.e. coconut oil)
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Not applicable
Duration of treatment / exposure:
104 wks
Frequency of treatment:
Daily, ad libitum in diet
Post exposure period:
Not applicable
Remarks:
Doses / Concentrations:
54 cal%
Basis:
nominal in diet
No. of animals per sex per dose:
39-40 per sex per dose
Control animals:
other: yes, 60 cal% soybean oil
Details on study design:
No details reported
Positive control:
No
Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS/MORTALITY: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Regularly

FOOD CONSUMPTION (if feeding study): Yes
- Time schedule for examinations: Twice, after 30 and 104 wk during 14 and 4 d

Other: GROWTH RATE: Yes
- Number of animals: 10 males/females per group
- Time schedule for exmainations: First 100 wks

Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
- Heart, liver, kidneys, adrenals and any tissue which showed abnormally gross appearance were examined histologically. Tissue sections were stained with hematoxylin-azophloxin and Masson's-Goldner trichrome.
Other examinations:
None
Statistics:
Yes, but details not provided
Clinical signs:
no effects observed
Mortality:
no mortality observed
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY: The differences in life span and in the time after which 50% of the animals died (50% mortality) between the two groups were not significant (i.e. 50% mortality was reached in test group by 803 M/886 F days and in control group (soybean oil): 849 M/883 F; Mean life span: Test group - 798 M/819 F; Control group - 819 M/864 F).

GROWTH RATE: No differences in growth were observed

FOOD CONSUMPTION (g/animal/day): Food intake of both sexes after 30 and 104 wks were higher (i.e. 30 wks: 14.3 M/10.1 F; 104 wks: 13.6 M/11.9 F) compared to control group (30 wks: 12.7 M/9.4 F; 104 wks: 12.1 M/9.9 F). But this increase in food intake did not reflect in differences in growth


HISTOPATHOLOGY: NON-NEOPLASTIC: Overall the frequencies of liver and kidney diseases were not significantly different in the groups of the two sexes separately or together (Incidences of nephrosis in kidney was the highest observed non-neoplastic change, but these effects were not significant; test group: 18 M/13 F; control group: 14 M/6 F). Other non-neoplastic changes of kidney (pyelonephritis, necrocalcinosis), cortical hemorrhage of adrenals, atrophy of testes in males, hyperplasia of parathyroids (males), necrocalcinosis and fibrosis of heart, edema, lipidosis of lungs were sporadic.

HISTOPATHOLOGY: NEOPLASTIC (if applicable): The differences in total number of tumour bearers (in %, mixed population out of n = 40) between the dietary groups were not significant (test group: 30% M/38% F; control group (39% M/49% F). Incidences of adenoma of pitutary was highest but not significant with respect to control group (i.e. test group: 6 M/11 F; 10 M/0 F). Other neoplastic tumors of adrenals, uterus, mamry gland, lymphoreticular tissue were sporadic.

Relevance of carcinogenic effects / potential:
Dietary administration of coconut oil has low carcinogenic potential.
Key result
Dose descriptor:
NOAEL
Effect level:
54 other: cal%
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects, including neoplastic and non-neoplastic changes in organs
Key result
Critical effects observed:
no

None

Conclusions:
Under the test conditions, dietary administration of 54 cal % of the substance to wistar rats of either sex, for entire life span, has low carcinogenic potential.
Executive summary:

A study was conducted to determine the long-term carcinogenic effects of the constituent 'glycerides, C8-18 and C18-unsatd.' (as coconut oil) in a life-span experiment in rats. 54 calorie (cal) % of the substance was fed through diet to 39-40 male and female Wistar rats per group during their entire life span (104 wks). No significant differences in the mean life span were found between groups. Considering the total population (males and females), the number of tumour bearers did not differ significantly. Parathyroid hyperplasia and necrocalcinosis in the heart were found mainly in males with slightly higher incidence in the coconut oil group in comparison with the control (soybean oil). Other pathological changes were randomly distributed among the different groups. However, the nature and incidence of these histopathological changes including tumours could not be related to the type of dietary fat or fatty acids used. Hence, under the test conditions, dietary administration of 54 cal% of the substance to wistar rats of either sex, for entire life span, has low carcinogenic potential (Vles and Gottenbus, 1972).

Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Reason / purpose for cross-reference:
reference to other study
Qualifier:
no guideline followed
Principles of method if other than guideline:
This study was designed to determine whether the constituent tocopherol would reduce the incidence of pulmonary tumorigenesis in the more susceptible A/J mice to an incidence similar to that of ddY mice. Five animals per group were killed after 28 weeks to determine the effect of tocopherol on oxidative stress on the pulmonary nuclei.
GLP compliance:
not specified
Species:
mouse
Strain:
other: A/J
Sex:
male
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
40 wks
Frequency of treatment:
Daily
Post exposure period:
Not applicable
Remarks:
Doses / Concentrations:
25.5 (control), 550 mg/kg bw/day
Basis:
nominal in diet
No. of animals per sex per dose:
15 males/dose
Control animals:
other: Yes, control diet, containing 25.5 mg/kg tocopherol was fed to groups of 15 A/J and ddY male mice
Relevance of carcinogenic effects / potential:
No carcinogenic effects were observed in terms of incidence of spontaneous pulmonary tumorigenesis in A/J mice.
Key result
Dose descriptor:
NOEL
Effect level:
550 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: 53% lower incidence of pulmonary neoplasms and 60% less pulmonary neoplasm multiplicity compared to A/J controls (incidence of pulmonary neoplasms in A/J test animals was similar to that of ddY controls).
Key result
Critical effects observed:
no

Details of results:

- No significant difference was observed in body weights between A/J mice of the test and control groups.

- At 28 weeks, the reduced nicotinamide adenine dinucleotide diphosphate (NADPH)-driven active oxygen generation, the nuclear thiobarbituric acid reactive substances (TBARS), and DNA single strand breaks were statistically significantly greater in A/J control animals than in ddYcontrol animals; dosing of A/J mice with tocopherol decreased the amount of TBARS and DNA single strand breaks compared to A/J controls.

- Nuclear alpha-tocopherol concentrations in A/J controls were significantly decreased compared to ddY controls; tocopherol increased the concentrations.

- A/J test animals had a 53% lower incidence of pulmonary neoplasms and 60% less pulmonary neoplasm multiplicity as compared to A/J controls; the incidence of pulmonary neoplasms in A/J test animals was similar to that of ddY controls.

- The plasma alpha-tocopherol concentration in A/J controls was 45% lower than that of ddY controls, and the plasma alpha-tocopherol concentration in tumor-bearing A/J controls was significantly decreased compared to that in nontumor-bearing A/J controls. Plasma alpha-tocopherol concentrations of test A/J animals was increased 140% compared to A/J controls.

Conclusions:
Under the test conditions, the substance was found to reduce incidence of spontaneous pulmonary tumorigenesis in A/J mice.
Executive summary:

A study was conducted to determine whether the constituent alpha-tocopherol would reduce the incidence of pulmonary tumorigenesis in the more susceptible A/J mice to an incidence similar to that of ddY mice A/J mice. A group of 15 male A/J mice were fed chow containing 550 mg/kg bw of the substance and groups of 15 A/J and ddY male mice were fed control diet (which contained 25.5 mg/kg bw of the substance) for 40 weeks. Five animals per group were killed after 28 weeks to determine the effects of the substance-induced oxidative stress on the pulmonary nuclei. The remaining 10 animals per group were killed at the termination of dosing. No significant difference was observed in body weights between A/J mice of the test and control groups. At 28 weeks, the reduced nicotinamide adenine dinucleotide diphosphate (NADPH)-driven active oxygen generation, the nuclear thiobarbituric acid reactive substances (TBARS), and DNA single strand breaks were statistically significantly greater in A/J control animals than in ddYcontrol animals. Dosing of A/J mice with the substance decreased the amount of TBARS and DNA single strand breaks compared to A/J controls. Nuclear test substance concentrations in A/J controls were significantly decreased compared to ddY controls. A/J test animals had a 53% lower incidence of pulmonary neoplasms and 60% less pulmonary neoplasm multiplicity as compared to A/J controls; the incidence of pulmonary neoplasms in A/J test animals was similar to that of ddY controls. The plasma concentration of the substance in A/J controls was 45% lower than that of ddY controls, and that in tumor-bearing A/J controls was significantly decreased compared to that in nontumor-bearing A/J controls. Plasma concentrations of the substance in test A/J animals was increased 140% compared to A/J controls. Hence, under the test conditions, the substance was found to reduce incidence of spontaneous pulmonary tumorigenesis in A/J mice (Fiume, 2002).

Endpoint:
carcinogenicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Carcinogenic effects of tocopheryl acetate were evaluated in a chronic repeated dose toxicity study in rats.
Species:
rat
Strain:
other: Charles River CD rats
Sex:
male/female
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
104 weeks (2 years)
Frequency of treatment:
Daily
Post exposure period:
Not applicable
Remarks:
Doses / Concentrations:
0, 500, 1000, or 2000 mg/kg bw/day
Basis:
nominal in diet
No. of animals per sex per dose:
60 animals/sex/dose
Control animals:
yes, plain diet
Relevance of carcinogenic effects / potential:
No significant difference in tumour incidence between control and treated animals could be attributed to alpha-tocopheryl acetate.
Key result
Dose descriptor:
NOAEL
Effect level:
2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: tumour incidence did not reveal any neoplastic effects of treatment
Key result
Critical effects observed:
no

- In both sexes, there were indications of an inverse relationship between dosage and incidence of mammary fibro-adenomas, but this effect was statistically significant only in females.

- No significant difference in tumour incidence between control and treated animals could be attributed to alpha-tocopheryl acetate.

- The distribution of pituitary adenomas displayed no related trend.

- Small incidences of liver cell tumours and of tumours of the biliary epithelium were scattered across the groups without relation to dosage or to treatment.

Conclusions:
Under the test conditions, the NOAEL of the substance for carcinogenic effects was 2,000 mg/kg bw/day, based on the lack of significant difference in tumour incidence between control and treated animals.
Executive summary:

The carcinogenic effects of the constituent dl-alpha-tocopheryl acetate were investigated in a chronic repeated dose toxicity study. Groups of 60 male/female Charles River CD rats, were fed a diet supplemented with the substance at dose levels of 500, 1000, or 2000 mg/kg bw/day for 104 weeks. The tumor incidences and group distribution of tumors along with systemic toxicity were monitored at intervals. Systemic effects have been reported in section 7.5.1 under “UM, Total tocopherol (dl-alpha-tocopheryl acetate) (200-412-2), 104 weeks, Repeated dose toxicity: oral (rat), KL2, JECFA, 1987" study. In both sexes, there were indications of an inverse relationship between dosage and incidence of mammary fibro-adenomas, but this effect was statistically significant only in females. Negative neoplastic response was interpretated from the group distribution of tumours. The number of neoplasms was also similar for all groups after 52 and 104 weeks of treatment. The distribution of pituitary adenomas displayed no related trend. Small incidences of liver cell tumours and of tumours of the biliary epithelium were scattered across the groups without relation to dosage or to treatment. Under the test conditions, the NOAEL of the substance for carcinogenic effects was 2,000 mg/kg bw/day, based on the lack of significant difference in tumour incidence between control and treated animals (JECFA, 1987).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Carcinogenicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Carcinogenicity: via dermal route

Link to relevant study records
Reference
Endpoint:
carcinogenicity: dermal
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Reason / purpose for cross-reference:
reference to other study
Qualifier:
no guideline followed
Principles of method if other than guideline:
The tumorogenic and co-carcinogenic (tumor promoting) effects of squalene were evaluated in two dermal application (skin painting) studies.
GLP compliance:
not specified
Species:
mouse
Strain:
other: C57B1 and C57BR mice
Sex:
not specified
Route of administration:
dermal
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
14 wks
Frequency of treatment:
three times weekly
Post exposure period:
Not applicable
Remarks:
Doses / Concentrations:
Undiluted squalene (i.e. 100% solution)
Basis:
nominal conc.
No. of animals per sex per dose:
Not reported
Control animals:
not specified
Key result
Dose descriptor:
NOEL
Effect level:
100 other: %
Based on:
test mat.
Sex:
not specified
Basis for effect level:
other: Absence of skin tumours
Key result
Dose descriptor:
conc. level:
Effect level:
100 other: %
Based on:
test mat.
Sex:
male
Basis for effect level:
other: plays a protective role against the tumour inducing property of 0.3% 3-methylcholanthrene (3-MCA) in mouse skin
Key result
Critical effects observed:
no

In the second skin-painting study (co-carcinogen study), each form (i.e. fresh and aged) of the squalene was determined to be inactive as a co-carcinogen. Further, when 0.3% 3-MCA in Squalene was “aged” by being left to stand for four weeks in the open air at 37°C, 3-MCA lost its carcinogenic effect on mouse skin.

Conclusions:
Under the test conditions, the undiluted substance was found to be non-tumourigenic to C57B1 and C57BR mice skin and play protective role against carcinogenic hydrocarbon, 0.3% 3-methylcholanthrene (3-MCA).
Executive summary:

The tumourigenic and co-carcinogenic effects of the constituent squalene were evaluated in two dermal application (skin painting) studies. For the carcinogenic study, undiluted forms of both freshly purified and “aged” squalene (compound that had been exposed to open air at 37°C for four weeks) were painted three times weekly for 14 weeks on the backs of C57B1 and C57BR mice. No skin tumors developed, and the substance was assessed to be non-tumorogenic to skin. Similar procedure as above was followed for the co-carcinogenic/tumour promoting study. Squalene in combination with 0.3% 3-methylcholanthrene (3-MCA), was used as the tumour inducer. Each form (i.e. fresh and aged) of the substance was determined to be inactive as a co-carcinogen. When 0.3% 3 -MCA in the substance was “aged” by being left to stand for four weeks in the open air at 37°C, 3-MCA lost its carcinogenic effect on mouse skin. Under the test conditions, the substance was not only found to be non-tumourogenic to skin, it was also assessed to play a protective role against hydrocarbon carcinogens (CIR, 1982).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Justification for classification or non-classification

Considering the evidence presented above, it is concluded that ‘Oils, vegetable, deodorizer distillates’ do not have carcinogenic potential and therefore does not warrant any classification according to CLP Regulation EC/1272/2008.

Additional information

There is no published information on the carcinogenicity of the test substance ‘Oils, vegetable, deodorizer distillates’. However, data is available for some of its constituents (see also table above):

·        Glycerides with chain lengths between C8 -18, including C18 -unsatd. were tested for carcinogenicity in rat (lifetime) at a dose representing 54% of the calories in diet (Vles and Gottenbus, 1972). There were no significant differences in lifespan, growth or food consumption between rats exposed to coconut oil, butter fat or soybean oil (hydrogenated or non-hydrogenated). Pathological changes were randomly distributed across the groups and considering the total population, the number of tumour bearing animals was comparable between groups. Additionally, other glycerides with unbranched fatty acids of overall longer chain lengths in the range of C16 -18, including C18 -unsatd. revealed non-promoting effect on chemically-induced mammary carcinogenesis in rats (Sylvester et al. 1986; Sundram et al. 1989). Furthermore, the‘glycerides’ (in the form of triglycerides of olive oil, corn oil, sunflower oil, etc.) are also well known to be frequently used as vehicles in toxicity studies following international testing guidelines (e.g. OECD) for the evaluation of repeated dose toxicity, carcinogenicity or reproductive/developmental toxicity of chemical substances, without any apparent adverse effects (CIR, 1997 and 2001b). They are also found to inhibit tumor promotion induced by standard promoters.

·        Numerous carcinogenicity studies have been conducted on fatty acids and their salts (in particular oleic acid), a number of which are already assessed in CIR (1987) and HERA (2002) reports. Although differences compared to controls were observed in individual trials, these are generally reported to be within normal background variation. Several fatty acids (stearic acid, oleic acid and sodium palmitate) are Generally Recognised as Safe (GRAS). Also, fatty acids as a group are permitted as direct food additives (HERA, 2002). Overall, fatty acids are not expected to be carcinogenic.

·        Tocopheryl acetate did not reveal any neoplastic effect when tested in a 2 -year carcinogenicity study in rats. Studies conducted in mice and rats suggested that tocopherols may, in some cases, have antitumourigenic properties. It is to be noted that tocopherols have often been used as a negative control in tumour promotion studies (Fiume, 2002).

·        No specific information could be found for sterols and sterol esters. However, given their long history of safe use in nutritional and cosmetic applications, no carcinogenic activity is expected.

·        The carcinogenicity of squalene was evaluated in two dermal application (skin painting) studies in mice, as summarised in CIR (1982). Results were not consistent between studies; some of the studies indicated potential effects whereas others showed a protective role against carcinogens. Overall, based on its history of safe use in nutritional and cosmetic applications, squalene is not expected to be carcinogenic under normal and foreseeable use conditions.

Moreover, predictions for ‘oils, vegetable, deodorizer distillates’(based on the main constituents) from the Danish QSAR database and Toxtree (v.1.6) model did not reveal any structural alerts for carcinogenicity.