Soaps, stocks, vegetable-oil, acidulated

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Studies conducted on different constituents of the substance (glycerides, fatty acids, tocopherols and squalene) indicated that these substances are not reproductive toxicant.

Link to relevant study records

Reference

Reference 1

Endpoint:

screening for reproductive / developmental toxicity

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

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

A 90 d oral repeated dose study was conducted in F344/N rat to evaluate the sub-chronic toxicity of castor oil in which reproductive parameters were also screened. Rats (10/sex/group) were treated for 13 weeks with 0, 0.62, 1.25, 2.5, 5.0 or 10% castor oil mixed in diet. Apart from the standard sub-chronic toxicity examinations, sperm count, motility and morphology were evaluated at necropsy and vaginal cytology during the week preceding necropsy. Male and female gonadal weights were also determined with gross pathology and histopathology of reproductive organs at termination.

GLP compliance:

yes

Remarks:

FDA Good Laboratory Practices Regulations (21 CFR 58)

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Simonsen Laboratories, Gilroy, CA
- Age at study initiation: 6 wk
- Fasting period before study: No
- Housing: 5 per cage in polycarbonate cages lined with heat-treated hardwood chips and covered with polyester filter sheets. The cages were stored on stainless steel racks equipped with an automatic watering system
- Diet: NIH 07; available ad libitum
- Water: Ad libitum
- Acclimation period: 14 d
- Other: Feed hoppers in the animal cages were changed twice weekly


ENVIRONMENTAL CONDITIONS
- Temperature: 68-76°F
- Humidity: 42-72%
- Air changes: 10/h
- Photoperiod: 12 h dark/12 h light

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Method of mixing: Formulated diets were prepared by blending the appropriate amount of castor oil with a small quantity of feed to prepare a premix. The premix then was layered between the required amounts of feed in a twin-shell blender and blended for 15 min to achieve a uniform mix.
- Storage temperature of food: Stored for no longer than 3 weeks at 5°C
- Stability under test conditions: 0.5% dose level is stable for at least 21 d when stored in the dark at 5°C and for 3 d when stored open to air and light in a rodent cage.

Details on mating procedure:

No mating performed

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Determined by HPLC at the study and analytical chemistry laboratories in duplicates. The results of the analyses for all dose mixtures given to the animals ranged from 97% to 106% of the target concentrations.

Duration of treatment / exposure:

13 wk or 90 d

Frequency of treatment:

Daily

Details on study schedule:

None

Remarks:

Doses / Concentrations:
0, 0.62, 1.25, 2.5, 5.0 or 10% in diet
Basis:
actual ingested

No. of animals per sex per dose:

10 rats per sex per dose

Control animals:

yes, plain diet

Details on study design:

None

Positive control:

Not applicable

Parental animals: Observations and examinations:

All routine examinations were performed including body weight, food consumption, hematology and clinical chemistry. For details see section 7.5.1: Repeated dose toxicity-oral; Endpoint study record: Castor oil (232-293-8), Irwin, 1992.

Oestrous cyclicity (parental animals):

Yes (at 12 wk)

Sperm parameters (parental animals):

Testis weight, epididymis weight, sperm count, motility and morphology were evaluated at necropsy (termination of study)

Litter observations:

Not examined

Postmortem examinations (parental animals):

Following reproductive organs were examined grossly and histologically apart from routine examinations:
Epididymis/seminal vesicles/prostate/testes or ovaries/uterus

- Complete histopathology examinations were conducted on all rats from the control and 10% dose groups

Postmortem examinations (offspring):

Not examined

Statistics:

Statistically analyzed within each sex by one-way Analysis of Variance tests, followed by Dunnett's t-test for pair-wise comparisons (p < 0.05)

Reproductive indices:

No data

Offspring viability indices:

Not calculated

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

not examined

No significant changes were noted for male reproductive endpoints, including sperm count and motility, and no changes were observed in the length of female estrous cycles. No significant changes were noted in male and female gonadal organs at necropsy except there was a slight decrease in epididymal weight (6-7%) which occurred in the middle- and high-dose groups, but this was not dose-related.

Key result

Dose descriptor:

NOAEL

Effect level:

> 10 other: % in diet (i.e. ca. 5,800 mg/kg bw/d) (male/female)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant changes for male reproductive endpoints, including sperm count and motility, and no changes in length of female estrous cycles

Key result

Critical effects observed:

no

Not applicable

Key result

Dose descriptor:

NOAEL

Remarks on result:

not measured/tested

Reproductive effects observed:

not specified

None

Conclusions:

Under the conditions of this study, the NOAEL of castor oil for reproductive toxicity screening in rats was determined to be 10% in diet (i.e. 5800 mg/kg bw/day based on actual feed consumption and body weight data).

Executive summary:

A 90 d oral repeated dose study was conducted in F344/N rats to evaluate the sub-chronic toxicity of the constituent castor oil in which reproductive parameters were also screened. Rats (10/sex/group) were exposed for 13 weeks to 0, 0.62, 1.25, 2.5, 5.0 or 10% castor oil mixed in diet. Apart from the standard sub-chronic toxicity examinations, sperm count, motility and morphology were evaluated at necropsy and vaginal cytology during the week preceding necropsy. Male and female gonadal weights were also determined with gross pathology and histopathology of reproductive organs at termination. No significant changes were noted in a screening for male reproductive endpoints, including sperm count and motility, and no changes were observed in the length of female estrous cycles. No significant changes were noted in male and female gonadal organs at necropsy except there was a slight decrease in epididymal weight (6-7%) which occurred in the middle- and high-dose groups, but this was not dose-related. Under the conditions of this study, the NOAEL of castor oil for reproductive toxicity screening in rats was determined to be 10% in diet (i.e. 5800 mg/kg bw/day based on actual feed consumption and body weight data) (Irwin, 1992).

Reference 2

Endpoint:

screening for reproductive / developmental toxicity

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

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

A 13 wks (90 d) oral combined repeated dose and reproduction/developmental screening study was conducted in Sprague Dawley rat to evaluate the reproductive parameters. After 10 weeks of treatment with 15% crude palm oil, 10 males and 20 females (15 - 16 weeks of age) were mated for 18 d. The females were then allowed to produce young. Maternal bodyweight and reproductive parameters (e.g. % implantation, % surviving young, % embryo loss, ...) were recorded. At 5 weeks of age, the young were sacrificed. One male and 2 females from each litter was sacrificed. Liver and kidneys were weighed. For 5 males and 5 females selected randomly, these organs were examined microscopically.

GLP compliance:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Own breeding
- Age at study initiation: ca. 6 weeks
- Weight at study initiation: 150 g
- Housing: 5 per sex per cage
- Diet (e.g. ad libitum): 40% wheat, 20% maize, 12% fish meal, 7% blood powder, 15% oil and 6% vitamin/minerals complement
- Water (e.g. ad libitum): yes

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): every 4 - 5 d
- Storage temperature of food: 4°C

Details on mating procedure:

After 10 weeks of treatment, 10 males and 20 females (15 - 16 weeks of age) were mated for 18 d. The females were then allowed to produce young.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Duration of treatment / exposure:

13 wks or 90 d

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
15%
Basis:
nominal in diet

No. of animals per sex per dose:

10 males, 20 females for the reproduction screening

Details on study design:

No data

Positive control:

No data

Parental animals: Observations and examinations:

REPRODUCTION SCREENING:
Maternal bodyweight and reproductive parameters (e.g. % implantation, % surviving young, % embryo loss etc) were recorded. At 5 weeks of age, the young were sacrificed. One male and 2 females from each litter was sacrificed. Liver and kidneys were weighed. For 5 males and 5 females selected randomly, these organs were examined microscopically.

Postmortem examinations (offspring):

Microscopic pathology of liver and kidney of young rats were performed from the reproduction screening at 35 d of age.

Statistics:

No data

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Reproductive performance:

no effects observed

During the reproductive screening study, no effects on maternal bodyweight evolution, reproductive parameters, pup liver and kidney weights, and pup liver and kidney histopathology

Key result

Dose descriptor:

NOAEL

Effect level:

> 15 other: % in diet (i.e., equivalent to 17,000 - 7,000 mg/kg bw/day as the bodyweight of animals increased regularly over the course of the study)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant toxicity on maternal rats or pups

Key result

Critical effects observed:

no

During the reproductive screening study, no effects were observed on maternal body weight, reproductive parameters, pup liver and kidney weights, and pup liver and kidney histopathology.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

15 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no toxicologically significant effects

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

None

Conclusions:

Under the conditions of this study, the NOAEL of the substance can be considered to be 15% in diet, as no significant toxicity were observed on maternal animals and pups.

Executive summary:

A combined repeated and reproductive screening study was conducted to determine the effect of the constituent ‘glycerides, C16-18 and C18-unsatd.’ (as crude palm oil) on rats when administered for 13 weeks at 15% in diet. Results were compared to those obtained with heated palm oil, crude/heated soy oil, crude/heated peanut oil, or crude/heated sunflower oil at the same concentration. Clinical signs and bodyweight were recorded throughout the study. After 13 weeks, hematology, clinical chemistry and urinalysis parameters were assessed, as well as gross and microscopic pathology. After 10 weeks of treatment, 10 males and 20 females were mated for 18 d. Maternal bodyweight and reproductive parameters were recorded. At 5 weeks of age, the young were sacrificed. Liver and kidneys weights were recorded and these organs were examined microscopically. The substance did not show any adverse effects on male and female rats compared to other crude or heated vegetable oils when administered for 13 weeks at 15% in diet. Furthermore, no signs of toxicity were observed on maternal rats or pups in the follow-up reproductive screening trial. Under the conditions of this study, the NOAEL of the substance can be considered to be 15% in diet as no significant toxicity were observed on maternal rats and pups (Coquet, 1977).

Reference 3

Endpoint:

multi-generation reproductive toxicity

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:

Multigeneration breeding studies in rats were conducted with the constituent red palm oil according to the protocol used in earlier studies on unconventional oils (Rukmini, 1988 and Rukmini, 1990) according to the guidelines of the FDA/WHO/DGHS safety evaluation protocol (FDA, 1970).

Groups of 24 (12 male and 12 female) inbred weanling albino rats were given a 20% protein diet, adequate in all nutrients and containing 10% of either groundnut oil (GNO; controls), red palm oil (RPO), refined, bleached, deodorized palm olein oil (RBDPO), or hydrogenated vegetable oil (HVPO) with 30% mahua oil.

Bodyweight and food intake were recorded weekly for 15 weeks (100 - 120 days).

Propagation of the three generations was conducted as follows: F0 rats were mated twice to produce F1a and F1B litters. F1b rats were mated twice to produce F2a and F2b litters, and two matings of the F2b rats produced the F3a and F3b litters. The parameters recorded included fertility index or conception rate, sex ratio, mean weaning weight, pre-weaning mortality, number of days from introduction of mating, behaviour of pups and adults.

GLP compliance:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

The rats were kept in galvanized aluminium cages at a constant temperature of 25°C, with 50-60% relative humidity and a 12 h light/dark cycle. The rats were given a 20% protein diet adequate in all nutrients. Food and water were given ad libitum.

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

Group 1 (control): 10% groundnut oil
Group 2: 10% red palm oil
Group 3: 10% bleached and deodorized palm olein
Group 4: 10% hydrogenated oil containing 30% of mahua oil

The substances were mixed in the diet.

Details on mating procedure:

12 males and 12 females in each group were mated after 100-120 d. The mated animals were allowed to be together for a period of 18-20 d. Mating behaviour was checked in the morning by looking for the capulatory plug in the bottom tray to confirm for pregnancy. After further confirmation by palpation and periodic checking for the increase in body weight in females, the mated animals were separated and housed in individual plastic cages.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

None

Duration of treatment / exposure:

3 generations

Frequency of treatment:

Daily

Details on study schedule:

The F0 rats were mated twice to produce the F1a and F1b litters. The F1b rats were mated twice to produce the F2a and F2b litters, and two matings of the F2b rats produced the F3a and F3b litters.

Once the female delivered, the litters were counted, sexed and weighted and return to the mother where they staid for the lactation period of 21 days. After the 21 days they were taken from the mothers, the number of female and males were recorded and their weight. Six male and six female pups of the F1a generation were killed after weaning. Gross pathology and organ weights were recorded. The mothers got to rest for 1 week and then again housed with the same males. This second mating produced F1b pups. After weaning the F1b pups were fed for 90 days, allowed to mature and bred to produce a second generation (F2b rats).

Remarks:

Doses / Concentrations:
10%
Basis:
nominal in diet

No. of animals per sex per dose:

12

Control animals:

yes

Details on study design:

The toxic or non-toxic responses from the reproduction studies were expressed in terms of indices that considered all stages from conception to weaning (as developed by Mirone L, Panzarella FP and Cerecodo LR, 1948. A new method of reporting data on reproduction and lactation in the mouse. Science,108:139).

Positive control:

None

Parental animals: Observations and examinations:

Bodyweight and food consumption, recorded weekly

Oestrous cyclicity (parental animals):

No data

Sperm parameters (parental animals):

No data

Litter observations:

The sex ratio, mean weaning weight, pre-weaning mortality and behaviour such as righting reflex (tested on Day 2 after birth), negative geotaxis (tested on Day 5 after birth), cliff drop aversion (tested on Day 5 after birth) and auditory startle response (tested on Day 12 after birth).

Postmortem examinations (parental animals):

The organs of all adults of each generation were weighed and the relative body weight were calculated.

Postmortem examinations (offspring):

Gross pathology and organ weight.

Statistics:

Analyses of variance was used to test the difference between groups for parametric data and the Chi-square test was used for non-parametric data.

Reproductive indices:

Fertility Index (FI)
Number of days from introduction for mating (IFM)
Behaviour of adults

Offspring viability indices:

Sex ratio
Mean weaning weight
Preweaning mortality
Behaviour of pups

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Reproductive performance:

effects observed, treatment-related

No significant differences were found in body weight gain between groups in any of the three generations. The food consumption and weight gain of rats fed crude palm oil was similar to that of the controls and they also showed an adequate growth and development. In the first generation 100% of matings resulted in pregnancies in both the first and the second mating. In the second generation 91.6% of dams conceived in both matings, and in the third generation 100% and 91.6% conceived in the first and second matings, respectively.

No abnormal behavioural or reflexological changes in any of the animals.

No significant changes attributable red palm oil were evident in the relative weights of the liver, heart, spleen, lungs, kidney, testes/ovaries or total body weight in the F0 generation rats.

Gross pathology indicates no adverse abnormalities in the organs.

Key result

Dose descriptor:

NOAEL

Effect level:

10 other: % in diet

Sex:

male/female

Basis for effect level:

other: no significant adverse effect were observed on any of the reproductive or toxicological parameters

Key result

Critical effects observed:

no

Key result

Dose descriptor:

NOAEL

Effect level:

10 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no toxicologically significant effects

Key result

Critical effects observed:

no

Clinical signs:

effects observed, treatment-related

Mortality / viability:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Mean litter size, birth and weaning weights were comparable and were not found to be significantly different in all the three generations of all groups.

The sex ratio at birth and weaning did not indicate any difference in the relative fitness of each sex with maturity. Pre-weaning mortality was higher in the first generation animals of both matings in all four groups, except for the second mating of the groups, which showed comparatively lower percentages. In the second mating of the second generation, the group showed a comparatively high percentage, and in the second mating of the third generation, percentages were higher for all groups. These mortalities are not likely to be treatment-related because the controls also had high pre-weaning mortality rates. Cannibalism was observed in some animals and the mortality of pups may be mainly due to this factor or to maternal negligence, rather than to the presence of any toxic substances that may have been excreted in milk or consumed by the pups.

There was no abnormality or unusual delay in conception or delivery in the groups.

No abnormal behavioural or reflexological changes in any of the animals.

No significant changes attributable to red palm oil were evident in the relative weights of the liver, heart, spleen, lungs, kidney, testes/ovaries or total body weight in the F1b or F2b generation rats.

Gross pathology indicates no adverse abnormalities in the organs.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

10 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no toxicologically significant effects

Key result

Critical effects observed:

no

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

10 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no toxicologically significant effects

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

None

Conclusions:

In this multigeneration study no adverse effects were observed on the reproductive or toxicological parameters studied. According to the author, these results indicate that the substance can be safely used as an edible oil.

Executive summary:

A three-generation study was conducted with groups of 12 male and 12 female rats fed 10% of either groundnut oil (controls), the constituent ‘glycerides, C16 -18 and C18 -unsatd.’ (as red palm oil), refined, bleached and deodorized palm olein or hydrogenated vegetable oil containing mahua oil through diet. Reproduction parameters including percentage conception, birth, weight, litter size, weanling weight, sex ratio at birth and weaning, preweaning mortality and number of days from introduction to mating, were recorded. Behavioural and reflexological tests were conducted on preweaning animals. No significant differences were recorded in any of the reproductive or toxicological parameters observed between the treated and control groups. According to the authors, these results indicate that the substance can be safely used as an edible oil (Manorama, 1993).

Reference 4

Endpoint:

two-generation reproductive toxicity

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 two generation reproductive toxicity study was conducted in Mongolian gerbils to assess the effects of dietary exposure to 8.75% w/w of the constituent palm kernel oil (PO). 5 couples of adult gebils with their sucklings (generation 1) were assigned to various diteary groups: palm kernel oil (8.75% w/w), sunflower seed oil (8.75% w/w) or cholesterol (0.2% w/w). The ensuing generations were put on the same diets as their parents with the exception of the cholesterol-containing diets which was reduced to 0.05%. After 4 months, 4 adults in each group were terminated and histopathological changes in liver and spleen were observed along with liver and serum cholesterol levels. In the second generation parameters such as frequency of litters, number of pups, mean litter size, mean weight and postnatal mortality were also observed.

GLP compliance:

not specified

Species:

other: gerbil

Strain:

other: Mongolian

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Adult gerbils and their sucklings (F1) were used at the initiation
- Housing: Housed in couples in standard laboratory cages with a wire mesh lid with food hopper. The bedding consisted of wood shavings.
- Diet (e.g. ad libitum): Diets manufactured by Hope Farms BV (Woerden, The Netherlands) were used. The test diet contained increased protein (26%), biotin, choline, inositol and vitamin E (50%) and cellulose (1%).
- Water (e.g. ad libitum): Water acidified with HCI (pH 3), ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature: 20°C
- Humidity: 50%
- Photoperiod: 10 h light/14 h dark

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

Not reported

Details on mating procedure:

No data

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Duration of treatment / exposure:

2 generations (specific period not mentioned)

Frequency of treatment:

Daily

Details on study schedule:

Not reported

Remarks:

Doses / Concentrations:
8.75% w/w in basic diet
Basis:
nominal conc.

No. of animals per sex per dose:

5 couples per group

Control animals:

yes, plain diet

Details on study design:

No data

Positive control:

Not applicable

Parental animals: Observations and examinations:

Not examined

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

Not examined

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in [F2] offspring: Frequency of litters, number of pups, mean litter size, mean weight at 6 months of age and postnatal mortality

Postmortem examinations (parental animals):

Not examined

Postmortem examinations (offspring):

SACRIFICE
- Four adult animals out of each dietary group of the F1 generation were sacrificed after 4 months.


HISTOPATHOLOGY / ORGAN WEIGHTS
- Tissues/organs examined: Liver and spleen
- Brief description on slide preparation: After fixation and paraffin embedding of parts of the liver and spleen, 4-µm thick sections were cut for analysis, followed by staining with hemaloxylin, eosin and Gomori's reticulin.


CLINICAL CHEMISTRY
- Total serum cholesterol and liver cholesterol content were also determined along with histopathological parameters in F1 generation after sacrifice

Statistics:

The Student-t test for unpaired samples was used to compare the variables of the different groups. For large differences in variance between the samples, the Wilcoxon test for unpaired samples was used. p < 0.05 was considered significant.

Reproductive indices:

Not reported

Offspring viability indices:

Not reported

No data

Key result

Dose descriptor:

NOAEL

Effect level:

8.75 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant treatment-related effect

Key result

Critical effects observed:

no

Key result

Dose descriptor:

NOAEL

Effect level:

8.75 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant treatment-related effect

Key result

Critical effects observed:

no

Clinical signs:

not examined

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

no effects observed

BODY WEIGHT (OFFSPRING): No significant difference compared to controls was observed (F1 and F2)


HISTOPATHOLOGY (F1): The livers and spleens of the animals on diets basal diet and palm kernel oil diet had a normal histology.


CLINICAL CHEMISTRY (F1):
- Serum cholesterol (mmol/L): Slight increase in PO fed group (3.9±0.5) in comparison with control (2.8±0.3).
- Liver cholesterol content: No significant difference was observed.


FREQUENCY OF LITTERS (F2): No significant effects were observed.


NUMBER OF PUPS (F2): No significant difference was observed (PO: 116 and basal diet (control): 146).


MEAN LITTER SIZE (F2): No significant difference was observed (PO: 4.7±1.7 and basal diet (control): 4.9±1.6).


POSTNATAL MORTALITY (F2): No significant mortality was observed in comparision to basal diet group ( PO: 4 % and basal diet (control): 3 %)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

8.75 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant treatment-related effect

Key result

Critical effects observed:

no

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

> 8.75 other: % w/w (i.e. ca. 4,375 mg/kg bw/day)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect on frequency of litters, mean litter size, total no. of newborns and no suckling death

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

None

Conclusions:

The test substance, exhibited no significant reproductive toxicity at 8.57% concentration level in a two generation study.

Executive summary:

A two generation reproductive toxicity study was conducted in Mongolian gerbils to assess the effect of the constituent ‘glycerides, C8-18 and C18-unsatd.’ (as palm kernel oil). Adult Mongolian gerbils (highly resistant to atherosclerosis) and their sucklings (first generation) were randomly assigned to two groups of 5 couples each (group 1: basal diet; group 2: 8.75% test substance in diet). Four months after starting the diets, four adult animals of each dietary group of the first generation were sacrificed and histopathological changes in liver and spleen were observed along with liver and serum cholesterol levels. In the second generation parameters such as frequency of litters, number of pups, mean litter size, mean weight, postnatal mortality were also observed. Both in the first and second generations, no significant differences of body weight were observed compared to controls. The liver and spleen of the animals of the first generation also had a normal histology. However, a slightly elevated serum cholesterol level was observed in the test substance fed group. No significant effect on frequency of litters, mean litter size, total no. of newborns, and no suckling death were found in the second generation of the test substance fed group compared to the basal dietary group. Hence, under the test conditions, the substance did not exhibit any significant reproductive toxicity at 8.57% in diet (Temmerman, 1988).

Reference 5

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Secondary literature source with limited details but used in the Integrated Laboratory Systems (ILS) assessment report

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Adult male albino Wistar rats exposed to β-sitosterol at 0.5 or 5 mg/kg bw/day (0.001 or 0.012 mmol/kg bw/day) for 16, 32 and 48 d respectively with a withdrawl period of 30 d were evaluated for fertility and reproductive parameters.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

No data

Route of administration:

subcutaneous

Vehicle:

not specified

Details on exposure:

No data

Details on mating procedure:

Not applicable

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

Low and high dose groups: Administered for 16, 32 and 48 d respectively.
Withdrawal group: Administered for 16, 32 and 48 d and then treatment was withdrawn for 30 d.

Frequency of treatment:

Daily

Details on study schedule:

Not applicable

Remarks:

Doses / Concentrations:
0.5 or 5 mg/kg bw/day (0.001 or 0.012 mmol/kg/day),
Basis:
nominal conc.

No. of animals per sex per dose:

10/dose

Details on study design:

No data

Positive control:

No data

Parental animals: Observations and examinations:

No data

Oestrous cyclicity (parental animals):

Not applicable

Sperm parameters (parental animals):

Not applicable

Litter observations:

Not applicable

Postmortem examinations (parental animals):

No data

Postmortem examinations (offspring):

Not applicable

Statistics:

No data

Reproductive indices:

Fertility, sperm concentrations and testicular weight were determined.

Offspring viability indices:

Not applicable

Clinical signs:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

not specified

Reproductive function: sperm measures:

effects observed, treatment-related

Reproductive performance:

effects observed, treatment-related

REPRODUCTIVE FUNCTION: SPERM MEASURES:
At 0.5 mg/kg bw/d β –sitosterol (0.00121 mmol/kg bw/d) there was significant decreased sperm concentrations after 48 d of treatment.
At 5 mg/kg bw/day (0.0121 mmol/kg bw/day) β –sitosterol, sperm concentrations were reduced after 16, 32, and 48 d of exposure.


REPRODUCTIVE PERFORMANCE
Fertility: At 5 mg/kg/day (0.0121 mmol/kg/day) β –sitosterol, fertility was reduced after 42 and 48 d of exposure.

Testicular weight:
At 0.5 mg/kg/day β –sitosterol ( 0.00121 mmol/kg/day) testicular weight was decreased after 32 and 48 d of treatment.
At 5 mg/kg testicular weight decreased in a time-dependent manner.

Withdrawal from treatment for 30 d did not restore sperm count or testicular weight.

Key result

Dose descriptor:

LOAEL

Effect level:

0.5 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: At 0.5 mg/kg bw/day there was a significant decrease in sperm concentration and testicular weight. At 5 mg/kg bw/day decreased fertility was observed in addition to other effects seen at low dose.

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

0.5 mg/kg bw/day (nominal)

System:

male reproductive system

Organ:

testes

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

not specified

Key result

Dose descriptor:

NOAEL

Remarks on result:

not measured/tested

Reproductive effects observed:

not specified

None

Conclusions:

Under the conditions of this study, the LOAEL of the substance in male rats was determined to be 0.5  mg/kg bw/day, based on decreased sperm concentration and testicular weight at lowest tested dose.

Executive summary:

A reproductive toxicity study was conducted through the sub-cutaneous route in male albino Wistar rats to assess the effect of the constituent β-sitosterol on reproductive performance in males. Adult male albino Wistar rats exposed to the substance at 0.5 or 5 mg/kg bw/day (0.001 or 0.012 mmol/kg bw/day) for 16, 32 and 48 d respectively (with a withdrawl period of 30 d) were evaluated for fertility and reproductive parameters. At 0.5 mg/kg bw/day, there was significantly decreased sperm concentrations and decreased testicular weight. At 5 mg/kg bw/day, fertility was reduced in addition to effects observed at lower dose. Under the conditions of this study, the LOAEL of the substance in male rats was determined to be 0.5  mg/kg bw/day, based on decreased sperm concentration and testicular weight at lowest tested dose (Tice, 1997).

Reference 6

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Secondary literature source with limited details, but used in the Australia New Zealand Food Authority (ANZFA) assessment of phytosterol esters, and followed GLP

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

A two generation reproductive toxicity study was conducted in Wistar rats to assess the dietary effects of 0, 1, 2 and 5% of the constituent plant sterol; equivalent to 0, 500–1300 mg/kg bw/day (low dose), 1000-2600 mg/kg bw/day (mid-dose) and 2800-4400 mg/kg bw/day (high dose)). Treatments began in F0 animals 10 weeks before mating and continued until weaning (3 weeks post parturition) when F0 males were sacrificed. F0 females were maintained for another 3 weeks and then sacrificed. During this entire period, various parameters were examined. Selected pups of F1 generation were treated with plant sterols in diet at the same levels as their F0 parents for 10 weeks pre-mating. The treatment, observation and sacrifice schedule followed that of the F0 generation. F1 and F2 litters were examined at 1, 4, 7, 14 and 21 d (weaning).

GLP compliance:

yes

Remarks:

Lab name not reported in the ANZFA risk analysis report

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld

Route of administration:

oral: feed

Vehicle:

not specified

Details on exposure:

Test substance intake was monitored:
At pre-mating and gestation sterol doses in males and females of F0 and F1 generation were in the ranges:
500-1300 mg/kg bw/day (low dose)
1000-2600 mg/kg bw/day (mid-dose)
2800-4400 mg/kg bw/day (high dose)

At 2 week lactation in females there was a slight increase in the sterol levels:
1700-1800 mg/kg bw/day (low dose)
3400-3500 mg/kg bw/day (mid-dose)
8500-9100 mg/kg bw/day (high dose).

Details on mating procedure:

No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

2 generations (Approx. 35 weeks)

Frequency of treatment:

Daily

Details on study schedule:

- F1 parental animals not mated until 10 wk after selected from the F1 litters.
- Selection of parents from F1 generation when pups were 6 wk of age.

Remarks:

Doses / Concentrations:
1, 2 and 5 % (made up from 1.6, 3.2 and 8.0% test material; equivalent to 500 – 1300 mg/kg bw/day (low dose), 1000 -2600 mg/kg bw/day (mid-dose) and 2800-4400 mg/kg bw/day (high dose)
Basis:
nominal in diet

No. of animals per sex per dose:

28 rats/sex/dose

Control animals:

yes, concurrent no treatment

Details on study design:

No data

Positive control:

Not applicable

Parental animals: Observations and examinations:

DAILY CLINICAL OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly (and for mated females on Days 0, 7, 14 and 21 of gestation, and on Days 1, 7, 14 and 21 post partum)

FOOD CONSUMPTION AND COMPOUND INTAKE: Weekly (not during mating)
Food effeciency: Pre-mating food efficiency (weeks 0-5 and 6-10 pre-mating, Days 0-21 gestation and Days 1-14 postpartum)

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

OTHER: Test substance intake

Oestrous cyclicity (parental animals):

In parent females, 3 weeks post weaning vaginal smears were performed to establish oestrus cycle length.

Sperm parameters (parental animals):

No data

Litter observations:

PARAMETERS EXAMINED
F1 and F2 litters were examined at 1, 4, 7, 14 and 21 d (weaning).
- Litter evaluation: On Days 4, 7, 14 and 21 post partum litters were evaluated for size, sex, still and live births, malformed offspring.
- Pup weight: On Days 1, 4, 7, 14 and 21 post partum

GROSS EXAMINATION OF DEAD PUPS:
- Weanling necropsy and histology of abnormal tissues (stillborns and intercurrent deaths)

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving F0 males were sacrificed 3 weeks post partum
- Female animals: All surviving F0 females were sacrificed 6 weeks post partum

GROSS NECROPSY & HISTOPATHOLOGY
- Necropsy and histology of F0 and F1 parental animals

Postmortem examinations (offspring):

SACRIFICE
- F1 offspring not selected as parental animals: 10 male and 10 female were subject to necropsy and tissue analysis.

Statistics:

No data

Reproductive indices:

Sexual maturation, fertility and reproductive performance were also monitored.

Offspring viability indices:

No data

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

BODY WEIGHT
Males: Mean body weights of F0 and F1 males at all dose groups was consistently lower than in controls. Differences in body weights were up to 6% in F0 and 8.5% in F1 animals. It was statistical significant at some time points in the highest dose group.
These differences were reflected in slight, and sometimes statistically significant differences between high dose males and controls in body weight change and food consumption and efficiency.

Females: In F0 females, there was slight (statistically insignificant) increase in body weight.

FOOD CONSUMPTION (PARENTAL ANIMALS)
Food consumption and efficiency: Slight and at some point statistically significant differences between high dose males and controls.

FERTILITY AND REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): Fertility and reproductive performance parameters were not significantly altered by sterol
treatment in either generation.

GROSS AND HISTOPATHOLOGYPATHOLOGY (PARENTAL ANIMALS): There were no consistent or dose related effects on organ weights or histopathology in either generation.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 2 800 - < 4 400 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Fertility, reproductive performance and litter data were not significantly altered by sterol treatment in either generation. There were no consistent or dose related effects on organ weights or histopathology in either generation.

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

There were also no dose-related changes in litter data.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 2 800 - < 4 400 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects

Key result

Critical effects observed:

no

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

>= 2 800 - < 4 400 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

None

Conclusions:

Under the conditions of this study, the NOAEL of the substance in rats was determined to be 5% (equivalent to 2,800-4,400 mg/kg bw/day in males and females) in diet based on lack of effects at the highest tested dose.

Executive summary:

A two generation reproductive toxicity study was conducted in Wistar rats to assess the dietary effects of the constituent plant sterols. 28 Wistar rats/sex/dose were administered dietary concentrations of 0, 1, 2 and 5% plant sterol; equivalent to 0, 500–1,300 mg/kg bw/day (low dose), 1,000-2,600 mg/kg bw/day (mid-dose) and 2,800-4,400 mg/kg bw/day (high dose) of the substance . Treatments began in F0 animals 10 weeks before mating and continued until weaning (3 weeks post parturition) when F0 males were sacrificed. F0 females were maintained for another 3 weeks and then sacrificed. During this entire period, various parameters were examined. Selected pups of F1 generation were treated with the substance in diet at the same levels as their F0 parents for 10 weeks pre-mating. The treatment, observation and sacrifice schedule followed that of the F0 generation. F1 and F2 litters were examined at 1, 4, 7, 14 and 21 d (weaning). Both in the first and second generations, no dose-related clinical observations were recorded. Fertility and reproductive performance parameters were not significantly altered by the substance treatment in either generation. There were also no dose related changes in litter data. There were no consistent or dose related effects on organ weights or histopathology in either generation. Slight significant differences of body weight were observed in males compared to controls. These differences were reflected in slight, and sometimes statistically significant differences between high dose males and controls in body weight change and food consumption and efficiency. Under the conditions of this study, the NOAEL of the substance in rats can be concluded as 5% (equivalent to 2,800-4,400 mg/kg bw/day in males and females) in diet based on lack of effects at the highest tested dose (ANZFA, 2001).

Reference 7

Endpoint:

screening for reproductive / developmental toxicity

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:

A reproductive toxicity study was conducted in Wistar rats to assess the effect of the constituent alpha-tocopherol. Fertility parameters such as pregnancy rates, vaiability of foetus, number of resorptions and developmental parameters such as effects on ossification and fetal weights were observed.

GLP compliance:

not specified

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on mating procedure:

Not reported

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

20 d prior to mating and during gestation

Frequency of treatment:

Daily

Details on study schedule:

Not reported

Remarks:

Doses / Concentrations:
0.75 (control), 7.5, or 75 mg/day alpha-tocopherol (equivalent to 2.14, 21.4 and 214.2 mg/kg bw/day, assuming the average rat body weight as 0.35 kg)
Basis:
nominal in diet

No. of animals per sex per dose:

10 male and female Wistar rats/dose

Control animals:

yes

Key result

Dose descriptor:

LOAEL

Effect level:

21 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: Decreased pregnancy rates (50% and 90% at 21 and 214 mg/kg bw/day respectively), few resorptions and mild cases of ossification were observed at both tested doses.

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

21 mg/kg bw/day (nominal)

System:

female reproductive system

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

not specified

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

214 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects observed

Key result

Reproductive effects observed:

yes

Lowest effective dose / conc.:

21 mg/kg bw/day (nominal)

Treatment related:

yes

Relation to other toxic effects:

not specified

Dose response relationship:

yes

Relevant for humans:

not specified

Details of results:

- The pregnancy rate for the animals of the 21 and 214 mg/kg bw/day groups were 90% and 50%, respectively.

- The number of viable fetuses was slightly less in the high dose group compared to controls.

- A few resorptions were observed for both test groups.

- Fetal weight was similar for all groups.

- Very mild cases of ossification retardation were observed. No abnormal fetuses were reported.

Conclusions:

Under the test conditions, the LOAEL of alpha- tocopherol for reproductive toxicity in rats was determined to be 21 mg/kg bw/day.

Executive summary:

A reproductive toxicity study was conducted in Wistar rats to assess the effect of the constituent alpha-tocopherol in rats. Groups of 10 male and female Wistar rats were fed a diet containing 0.75 (control), 7.5, or 75 mg/day of the substance (i.e. equivalent to 2.14, 21.4 and 214.2 mg/kg bw/day, assuming the average rat body weight as 0.35 kg) 20 d prior to mating and during gestation. Gravid animals were killed on Day 20 of gestation. All of the animals given control feed were gravid, but the pregnancy rate for the animals of the 21and 214 mg/kg bw/day groups were 90% and 50%, respectively. The number of viable fetuses was slightly less in the 214 mg/day group compared to controls. A few resorptions were observed for both test groups. Fetal weight was similar for all groups. Very mild cases of ossification retardation were observed. No abnormal fetuses were reported. Under the test conditions, the LOAEL of alpha- tocopherol for reproductive toxicity in rats was determined to be 21 mg/kg bw/day (Fiume, 2002).

Reference 8

Endpoint:

extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)

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

Principles of method if other than guideline:

The effects on reproductive indices were investigated on dietary exposure to 0, 0.002, 0.2, & 2% (i.e., approx. 1.02, 103 and 1028 mg/kg bw/day) d-alpha-tocopherol (polyethylene glycol) 1000 succinate (TPGS).

GLP compliance:

not specified

Justification for study design:

- The animals were mated on Day 112 of treatment to produce the F1a generation and on Day 175 to produce the F1b generation. The F0 animals were maintained on their respective diets for 265 d of treatment, then sacrificed and examined histopathologically.

Species:

rat

Strain:

other: Charles River CD

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not reported in the JECFA evaluation report

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

Not reported in the JECFA evaluation report

Details on mating procedure:

- The animals were mated on Day 112 of treatment to produce the F1a generation and on Day 175 to produce the F1b generation. The F0 animals were maintained on their respective diets for 265 d of treatment, then sacrificed and examined histopathologically.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not applicable

Duration of treatment / exposure:

265 d

Frequency of treatment:

Daily

Details on study schedule:

The animals were mated on Day 112 of treatment to produce the F1a generation and on Day 175 to produce the F1b generation. The F0 animals were maintained on their respective diets for 265 d of treatment, then sacrificed and examined histopathologically.

Remarks:

Doses / Concentrations:
0, 0.002, 0.2, & 2% (i.e., approx. 1.02, 103 and 1028 mg/kg bw/day, assuming 0.35 kg as average weight of adult rats and 18 g as daily food consumption)
Basis:
nominal in diet

No. of animals per sex per dose:

15/sex/dose

Control animals:

yes, plain diet

Details on study design:

Not reported in the JECFA evaluation report

Positive control:

Not reported in the JECFA evaluation report

Parental animals: Observations and examinations:

Clinical chemical and haematological parameters were observed at regular intervals

Oestrous cyclicity (parental animals):

Not reported in the JECFA evaluation report

Sperm parameters (parental animals):

Not reported in the JECFA evaluation report

Litter observations:

Not reported in the JECFA evaluation report

Postmortem examinations (parental animals):

The F0 animals were maintained on their respective diets to 265 d of treatment, then sacrificed and examined histopathologically.

Postmortem examinations (offspring):

Not reported in the JECFA evaluation report

Statistics:

Not reported in the JECFA evaluation report

Reproductive indices:

Reproductive indices such as mean gestation period, litter size, sex ratio, and mortality of pups or parents were observed

Offspring viability indices:

Not reported

Clinical signs:

no effects observed

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Reproductive indices (mean gestation period, litter size, sex ratio, and mortality of pups or parents) were unaffected by treatment.

Key result

Dose descriptor:

NOAEL

Effect level:

> 1 028 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects on reproductive indices, clinical chemical and haematological parameters

Key result

Critical effects observed:

no

Clinical signs:

not specified

Mortality / viability:

no mortality observed

Body weight and weight changes:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Not reported in the JECFA evaluation report

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 028 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects

Reproductive effects observed:

not specified

None

Conclusions:

Under the test conditions, the NOAEL of the substance was determined to be 2% (1028 mg/kg bw/day), based on lack of reproductive toxicity at the highest tested dose.

Executive summary:

The dietary effects on reproductive indices were investigated on exposure to d-alpha-tocopherol (polyethylene glycol) 1000 succinate (TPGS). Groups of 15 Charles River CD rats of each sex were fed diets containing the substance at concentrations of 0, 0.002, 0.2, or 2% (i.e. approx. 1.02, 102.8, 1028.4 mg/kg bw/day) for 265 d.The animals were mated on Day 112 of treatment to produce the F1a generation and on Day 175 to produce the F1b generation. The F0 animals were maintained on their respective diets to 265 d of treatment, then sacrificed and examined histopathologically. Reproductive indices (mean gestation period, litter size, sex ratio, and mortality of pups or parents) were unaffected by treatment. Clinical chemical and haematological parameters were normal in the F0 generation 10 d before terminal sacrifice. Under the test conditions, the NOAEL of the substance was determined to be 2% (1028 mg/kg bw/day), based on lack of reproductive toxicity at the highest tested dose (JECFA, 1987).

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

241 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no adverse effect observed

Additional information

There is no published information on the reproductive toxicity of ‘Soaps, stocks, vegetable oil, acidulated’. However, data is available for some of its constituents (see also table above):

·        Dietary exposure to 8.75% (i.e., approximately 4,375 mg/kg bw/day) glycerides with chain lengths of C8 - 18, including C18 -unsatd. was assessed in a two generation reproductive toxicity study in gerbils (Temmermans et al., 1988). No significant effect on frequency of litters, mean litter size, total number of newborns or death in weanling stage were found in the second generation. This is in line with findings from studies conducted with other glycerides having long chain lengths such as C16 -18, including C18 -unsatd. and C18 -unsatd. hydroxy (Irwin, 1992; Manorama et al., 1993; Coquet et al., 1977). Across all studies for glycerides, tested doses ranged from 8.75 to 15% in diet. No significant toxicity was seen at any of the doses tested. The highest oral NOAEL was 15% in diet which is equivalent to 7,000 to 17,000 mg/kg bw/day, from a 13 -week combined repeated dose and reproduction/developmental screening (feeding) study.

·        A number of studies have been conducted on individual fatty acids, as summarised in HERA (2002). At 15% in diet (ca. 7,500 mg/kg bw/day) for 10 - 16 weeks, oleic acid (C18) did not affect fertility in male rats but appeared to interfere with parturition and mammary gland development in females. The information is however insufficient to be able to verify and conclude on these results. A three-generation study with an oil containing 76% capric acid (C10) administered via diet did not produce any reproductive effects (Hendrich et al., 1993).

·        Tocopherols have been tested for reproductive toxicity in a series of studies (Tomassi and Silano, 1986 and Fiume, 2002). No adverse effects were observed.

·       Sterols: A 2 -generation reproduction study in rats fed up to 5% sterols in diet (ca. 4,400 mg/kg bw/day) for 10 weeks prior to mating then throughout gestation, lactation and weaning revealed no significant effects on F0 or F1 generations (ANZFA, 2001). Further, there were couple of studies with beta-sitosterol which revealed adverse effects on estrous cycle and male sperms (Tice, 1999) but these effects have not been reviewed further as the exposure route was not considered relevant under the foreseeable use conditions. Also, the beta-sitosterols are present at very low concentrations compared to other constituents without adverse effects.

·        No specific information could be found for squalene. However, given its long history of safe use in nutritional and cosmetic applications, reproductive toxicity is not expected.

Taken together, the above evidence suggests that ‘Soaps, stocks, vegetable oil, acidulated’ is not expected to be a reproductive toxicant.

Effects on developmental toxicity

Description of key information

Studies conducted on glycerides, fatty acids, tocopherols and squalene, indicate that these substances are not developmental toxicant.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Developmental toxicity/teratogenicity potential was observed in a two generation reproductive toxicity study after dietary administration of the constituent 15% partially hydrogenated soybean oil in Sprague Dawley rats.

Groups of 25 pairs of two generations of male and female rats were fed diets containing 15% of fresh partially hydrogenated soybean oil. F0 generation was exposed to the test material from weaning and F1 generation from conception. The first two litters of each generation were
permitted to be born naturally. During the third pregnancy of each generation, one-half of the females were sacrificed on Day 13 of gestation and inspected for early embryonic death. The remaining females were sacrificed on Day 21 of gestation, and the fetuses were examined for either skeletal or soft tissue abnormalities.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Diet: Semipurified rat diet, ad libitum

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): Weekly
- Mixing appropriate amounts with (Type of food): Semipurified rat diet
- Storage temperature of food: Under refrigeration

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not applicable

Details on mating procedure:

- Proof of pregnancy: Cornified cells and spermatozoa in vaginal smear; referred to as Day 0 of pregnancy

Duration of treatment / exposure:

F0 generation: From weaning and F1 generation: From conception

Frequency of treatment:

Daily ad libitum in diet

Duration of test:

Up to Day 13/21 of gestation or up to weaning of offsprings (see further details on study design for explanation)

Remarks:

Doses / Concentrations:
15%
Basis:
nominal in diet

No. of animals per sex per dose:

25 animals/sex

Control animals:

no

Details on study design:

The first two litters of F0 and F1 generations (i.e. F1a, F1b, F2a and F2b) were permitted to be born naturally. All F1a, F2a and F2b litters were discarded at weaning. F1b rats were allowed to grow for further mating. During the third pregnancy of each generation (i.e. F1c and F2c), one-half of the females were sacrificed on Day 13 of gestation and inspected for early embryonic death. The remaining females were sacrificed on Day 21 of gestation, and the fetuses were examined for either skeletal or soft tissue abnormalities.

Maternal examinations:

BODY WEIGHT: Weekly during the first 8 wks (after weaning)


FOOD CONSUMPTION: Weekly during the first 8 wks (after weaning)


POST-MORTEM EXAMINATIONS: Yes
- Organs examined: Heart, lung, stomach, liver, kidney, adrenals, small and large intestine, spleen, gonads, bladder, pancreas and mesenteric lymph nodes

Ovaries and uterine content:

The uterine content was examined after termination: Yes
Examinations included:
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes

Fetal examinations:

- Soft tissue examinations: Yes
- Skeletal examinations: Yes

Statistics:

The data were analyzed statistically by the Analysis of Variance and Chi-square method

Indices:

None

Historical control data:

No data

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
No evidence of maternal toxicity in both F0 and F1 generation

Key result

Dose descriptor:

NOAEL

Effect level:

15 other: % in diet

Basis for effect level:

other: no treatment-related effects

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No dead fetuses or any evidence of teratogenic effects

Key result

Dose descriptor:

NOAEL

Effect level:

15 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects on development of the fetuses

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

Table 1. Effect of partially hydrogenated soybean oil on development of rat fetus

 

Generation	F1c	F2c
No. of fetuses examined for soft-tissue defects	65	75
Fetuses with soft-tissue defects	0	0
No. of fetuses examined for skeletal defects	30	37
Fetuses with skeletal defects	0	1

Conclusions:

Under the conditions of this study, the NOAEL for maternal and developmental toxicity was considered to be 15% of the substance in diet.

Executive summary:

A study was conducted to evaluate the developmental toxicity/teratogenicity potential of the constituent 15% ‘glycerides, C8 -18 and C18 -unsatd.’ (as partially hydrogenated soybean oil) in Sprague Dawley rats. Groups of 25 pairs of two generations of male and female rats were fed diets containing 15% of the substance. The F0 generation was exposed to the substance from weaning and the F1 generation from conception. The first two litters of each generation were allowed to be born naturally. During the third pregnancy of each generation, one-half of the females were sacrificed on Day 13 of gestation and inspected for early embryonic death. The remaining females were sacrificed on Day 21 of gestation and fetuses were examined for either skeletal or soft tissue abnormalities. No evidence of any maternal or developmental toxicity (including teratogenicity) was observed in any of the generations. Under the conditions of this study, the NOAEL for maternal and developmental toxicity was considered to be 15% of the substance in diet (Nolen, 1972).

Reference 2

Endpoint:

developmental toxicity

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

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

A two generation reproductive toxicity study was conducted in gerbils to assess the effects of dietary exposure to 8.75% w/w palm kernel oil (PO).
Adult Mongolian gerbils (highly resistant to atherosclerosis) and their sucklings (first generation) were randomly assigned to two groups of 5 couples each (group 1: basal diet; group 2: 8.75% w/w PO in diet). In the second generation parameters such as frequency of litters, number of pups, mean litter size, mean weight, postnatal mortality were observed.

GLP compliance:

not specified

Limit test:

no

Species:

other: gerbil

Strain:

other: Mongolian

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Adult gerbils and their sucklings (F1) were used at the initiation
- Housing: Housed in couples in standard laboratory cages with a wire mesh lid with food hopper. The bedding consisted of wood shavings.
- Diet (e.g. ad libitum): Diets manufactured by Hope Farms BV (Woerden, The Netherlands) were used. The test diet contained increased protein (26%), biotin, choline, inositol and vitamin E (50%) and, cellulose (1%).
- Water (e.g. ad libitum): Water acidified with HCI (pH 3), ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature: 20°C
- Humidity: 50%
- Photoperiod: 10 h light/14 h dark

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

No data

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Details on mating procedure:

No data

Duration of treatment / exposure:

2 generations (specific period not mentioned)

Frequency of treatment:

Daily

Duration of test:

Up to 6 months of F2 generation

Remarks:

Doses / Concentrations:
8.75% w/w
Basis:
nominal in diet

No. of animals per sex per dose:

5 couples per group

Control animals:

yes, plain diet

Details on study design:

No data

Maternal examinations:

No data

Ovaries and uterine content:

Not examined

Fetal examinations:

The following parameters were examined in [F2] offspring: Frequency of litters, number of pups, mean litter size, mean weight at 6 months of age and postnatal mortality

Statistics:

The Student-t test for unpaired samples was used to compare the variables of the different groups. For large differences in variance between the samples, the Wilcoxon test for unpaired samples was used. p < 0.05 was considered significant.

Indices:

Litter frequency: calculated by dividing the total number of weeks during which the couples of gerbils within one dietary group were able to breed by the number of litters

Historical control data:

No data

Details on maternal toxic effects:

Maternal toxic effects:no data

Details on maternal toxic effects:
Not applicable

Key result

Dose descriptor:

NOAEC

Effect level:

> 8.75 other: % w/w

Based on:

test mat.

Basis for effect level:

other: no treatment-related effects

Key result

Abnormalities:

no effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:not examined

Details on embryotoxic / teratogenic effects:
Not applicable

Key result

Dose descriptor:

NOAEL

Effect level:

8.75 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects on development

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

OFFSPRING PARAMETERS:

 

BODY WEIGHT (OFFSPRING): No significant difference compared to controls was observed (F1 and F2)

FREQUENCY OF LITTERS (F2): No significant effects were observed.


NUMBER OF PUPS (F2): No significant difference was observed (PO: 116 and basal diet (control): 146).


MEAN LITTER SIZE (F2): No significant difference was observed (PO: 4.7±1.7 and basal diet (control): 4.9±1.6).


POSTNATAL MORTALITY (F2): No significant mortality was observed in comparison to basal diet group ( PO: 4% and basal diet (control): 3%)

Conclusions:

Based on the test conditions, the NOAEL for developmental effects in rats was determined to be 8.57% w/w.

Executive summary:

A two generation reproductive toxicity study was conducted in Mongolian gerbils to assess the effect of the constituent ‘glycerides, C8 -18 and C18 -unsatd.’ (as palm kernel oil). Adult Mongolian gerbils (highly resistant to atherosclerosis) and their sucklings (first generation) were randomly assigned to two groups of 5 couples each (group 1: basal diet; group 2: 8.75% w/w of the substance in diet). In the second generation parameters such as frequency of litters, number of pups, mean litter size, mean weight, postnatal mortality were observed. No significant effect on frequency of litters, mean litter size, total no. of newborns, mean body weight and no postnatal mortality were found in the second generation of the substance-treated group compared to the basal dietary group. Based on the test conditions, the NOAEL for developmental effects in rats was determined to be 8.57% w/w (Temmerman, 1988).

Reference 3

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Secondary literature source with limited details, but used in the Australia New Zealand Food Authority (ANZFA) assessment of phytosterol esters, and followed GLP

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Principles of method if other than guideline:

A two generation reproductive toxicity study was conducted in Wistar rats to assess the dietary effects of 0, 1, 2 and 5% plant sterol (made up from 0, 1.6, 3.2 and 8.0%; equivalent to 0, 500–1300 mg/kg bw/day (low dose), 1000-2600 mg/kg bw/day (mid-dose) and 2800-4400 mg/kg bw/day (high dose). Treatments began in F0 animals 10 weeks before mating and continued until weaning (3 weeks post parturition) when F0 males were sacrificed. F0 females were maintained for another 3 weeks and then sacrificed. During this entire period, various parameters were examined. Selected pups of F1 generation were treated with plant sterols in diet at the same levels as their F0 parents for 10 weeks pre-mating. The F1 and F2 litters were examined at weaning days 1, 4, 7, 14 and 21 for various parameters including developmental toxicity.

GLP compliance:

yes

Remarks:

Lab name not reported in the ANZFA risk analysis report

Limit test:

no

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld

Route of administration:

oral: feed

Vehicle:

not specified

Details on exposure:

Test substance intake was monitored:
At pre-mating and gestation sterol doses in males and females of F0 and F1 generation were in the ranges:
500-1,300 mg/kg bw/day (low dose)
1,000-2,600 mg/kg bw/day (mid-dose)
2,800-4,400 mg/kg bw/day (high dose)

At 2 week lactation in females there was a slight increase in the sterol levels:
1,700-1,800 mg/kg bw/day (low dose)
3,400-3,500 mg/kg bw/day (mid-dose)
8,500-9,100 mg/kg bw/day (high dose).

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Details on mating procedure:

No data

Duration of treatment / exposure:

2 generations

Frequency of treatment:

Daily

Duration of test:

Approx. 35 weeks

Remarks:

Doses / Concentrations:
1, 2 and 5 % (made up from 1.6, 3.2 and 8.0% test material; equivalent to 500 – 1,300 mg/kg bw/day (low dose), 1,000 -2,600 mg/kg bw/day (mid-dose) and 2,800-4,400 mg/kg bw/day (high dose)
Basis:
nominal in diet

No. of animals per sex per dose:

28 rats/sex/dose

Control animals:

yes, concurrent no treatment

Details on study design:

No data

Maternal examinations:

DAILY CLINICAL OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly (and for mated females on Days 0, 7, 14 and 21 of gestation and on Days 1, 7, 14 and 21 post partum)

FOOD CONSUMPTION AND COMPOUND INTAKE: Weekly (not during mating)
Food effeciency: Pre-mating food efficiency (weeks 0-5 and 6-10 pre-mating, Days 0-21 gestation and 1-14 postpartum)

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

OTHER: Test substance intake

Ovaries and uterine content:

No data

Fetal examinations:

PARAMETERS EXAMINED
F1 and F2 litters were examined at weaning days 1, 4, 7, 14 and 21
- Litter evaluation: On Day 4, 7, 14 and 21 post partum litters were evaluated for size, sex, still and live births, malformed offspring.
- Pup weight: On Day 1, 4, 7, 14 and 21 post partum

GROSS EXAMINATION OF DEAD PUPS:
- Weanling necropsy and histology of abnormal tissues (stillborns and intercurrent deaths)

Statistics:

No data

Indices:

No data

Historical control data:

No data

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
In F0 females, there was slight (statistically insignificant) increase in body weight.

Key result

Dose descriptor:

NOEL

Effect level:

>= 2 800 - < 4 400 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no treatment-related effects

Key result

Abnormalities:

no effects observed

Key result

Dose descriptor:

NOAEL

Effect level:

>= 2 800 - < 4 400 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects on development of F0 and F1 fetuses

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

None

Conclusions:

Under the conditions of this study, the NOEL of the substance for developmental toxicity in rats was determined to be 5% (equivalent to 2,800-4,400 mg/kg bw/day in males and females) in diet, based on lack of any morphological or pathological findings in pups of F0 and F1 generations.

Executive summary:

A two-generation reproductive toxicity study was conducted in Wistar rats to assess the effect of the constituent plant sterols and was also used to assess any developmental abnormality in pups. In the study, 28 Wistar rats/sex/dose were administered dietary concentrations of 0, 1, 2 and 5% plant sterol (made up from 0, 1.6, 3.2 and 8.0%; equivalent to 0, 500–1,300 mg/kg bw/day (low dose), 1,000-2,600 mg/kg bw/day (mid-dose) and 2,800 -4,400 mg/kg bw/day (high dose) of the substance . Treatments began in F0 animals 10 weeks before mating and continued until weaning (3 weeks post parturition) when F0 males were sacrificed. F0 females were maintained for another 3 weeks and then sacrificed. During this entire period, various parameters were examined. Selected pups of F1 generation were treated with the substance in diet at the same levels as their F0 parents for 10 weeks pre-mating. The treatment, observation and sacrifice schedule followed that of the F0 generation. The F1 and F2 litters were examined at weaning days 1, 4, 7, 14 and 21 for various parameters including developmental toxicity. Both in the first and second generations, no dose-related significant morphological or pathological findings were observed in pups of F0 and F1 generation with the substance at 5% of diet.  Under the conditions of this study, the NOEL of the substance for developmental toxicity in rats was determined to be 5.0% (equivalent to 2800 -4400 mg/kg bw/day in males and females) in diet, based on lack of any morphological or pathological findings in pups of F0 and F1 generations (ANZFA, 2001).

Reference 4

Endpoint:

developmental toxicity

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

Principles of method if other than guideline:

A developmental toxicity study was conducted in Wistar rats to assess the effect of the constituent alpha-tocopherol acetate.

GLP compliance:

not specified

Species:

rat

Strain:

Wistar

Route of administration:

oral: gavage

Vehicle:

corn oil

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

Not reported

Duration of treatment / exposure:

GD 6-15

Frequency of treatment:

Daily

Duration of test:

GD 6-20

Remarks:

Doses / Concentrations:
16, 74.3, 345 or 1,600 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

21, 23, 21, and 22 gravid albino Wistar rats for 16, 74.3, 345 or 1,600 mg/kg bw/day

Control animals:

yes, sham-exposed

Key result

Dose descriptor:

NOAEL

Effect level:

1 600 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: no treatment-related effects

Key result

Abnormalities:

no effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Key result

Dose descriptor:

NOAEL

Effect level:

1 600 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects on fetal development

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

None

Conclusions:

Under the test conditions, the NOAEL of the substance for developmental toxicity was determined to be 1,600 mg/kg bw/day.

Executive summary:

A study was conducted to evaluate the developmental toxicity potential of the constituent alpha-tocopherol acetate in Wistar rats. Groups of 21, 23, 21, and 22 gravid albino Wistar rats were dosed orally at 16, 74.3, 345, or 1,600 mg/kg bw/day of the substance in corn oil during Days 6 to 15 of gestation. Body weights were recorded on Days 0, 6, 11, 15, and 20 of gestation. The animals were killed on Day 20 of gestation and the fetuses were examined. The administration of up to 1600 mg/kg bw/day of the substance to pregnant rats had no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls. Under the test conditions, the NOAEL of the substance for developmental toxicity was determined to be 1,600 mg/kg bw/day (Fiume, 2002).

Reference 5

Endpoint:

developmental toxicity

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 teratogenic effects of the constituent d-alpha-tocopherol (polyethylene glycol) 1000 succinate (TPGS) were investigated upon dietary exposure to 0, 0.002, 0.2 and 2% (i.e. 1.02, 103 and 1,028 mg/kg bw/day, assuming 0.35 kg as average weight of adult rats and 18 g as daily food consumption) on Days 6 to 16 of gestation. On Day 20 of gestation, the dams were sacrificed, the uteri excised, and the number of implantation sites (live fetuses, dead fetuses, or resorption sites) were counted. All the fetuses were examined for gross anomalies; half were examined for soft-tissue abnormalities (Wilson technique) and half were examined for skeletal defects.

GLP compliance:

not specified

Species:

rat

Strain:

other: Charles River CD

Details on test animals or test system and environmental conditions:

Not reported in the JECFA evaluation report

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

Not reported in the JECFA evaluation report

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not applicable

Details on mating procedure:

Not reported in the JECFA evaluation report

Duration of treatment / exposure:

GD 6-16

Frequency of treatment:

Daily

Duration of test:

up to GD 20

Remarks:

Doses / Concentrations:
0, 0.002, 0.2, and 2% (i.e., approx. 1, 103 and 1,028 mg/kg bw/day)
Basis:
nominal in diet

No. of animals per sex per dose:

15 pregnant females/dose

Details on study design:

Not reported in the JECFA evaluation report

Maternal examinations:

Not reported in the JECFA evaluation report

Ovaries and uterine content:

On Day 20 of gestation, the dams were sacrificed, the uteri excised, and the number of implantation sites (live fetuses, dead fetuses, or resorption sites) were counted.

Fetal examinations:

All the fetuses were examined for gross anomalies; half were examined for soft-tissue abnormalities (Wilson technique) and half were examined for skeletal defects (stained using alizarin red).

Statistics:

Not reported in the JECFA evaluation report

Indices:

Not reported in the JECFA evaluation report

Historical control data:

Not reported in the JECFA evaluation report

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Not reported in the JECFA evaluation report

Key result

Dose descriptor:

NOEL

Effect level:

> 1 028 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no treatment-related effects

Key result

Abnormalities:

no effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
No differences were observed between controls and any of the treatment groups with respect to the parameters studied.

Key result

Dose descriptor:

NOAEL

Effect level:

1 028 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

None

Conclusions:

Under the conditions of the study, the NOEL of the substance for teratogenic and fetotoxic effects was determined to be >1,028 mg/kg bw/day, based on lack of developmental effects at the highest tested dose.

Executive summary:

The teratogenic/fetotoxic effects on dietary exposure to d-alpha-tocopherol (polyethylene glycol) 1000 succinate (TPGS) was determined in Charles River CD rats. Groups of 15 pregnant Charles River CD rats were administered the substance through diet at concentrations of 0, 0.002, 0.2, or 2% (i.e. approx. 1.02, 103 and 1,028 mg/kg bw/day) on Days 6 to 16 of gestation. On Day 20 of gestation, the dams were sacrificed, the uteri excised, and the number of implantation sites (live fetuses, dead fetuses, or resorption sites) were counted. All the fetuses were examined for gross anomalies; half of the fetuses were examined for soft-tissue abnormalities and remaining half were examined for skeletal defects. No differences were observed between controls and any of the treatment groups with respect to the parameters studied. Under the conditions of the study, the NOEL of the substance for teratogenic and fetotoxic effects was determined to be >1,028 mg/kg bw/day, based on lack of effects at the highest tested dose (JECFA, 1987).

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

241 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no adverse effect observed

Additional information

There is no published information on the developmental toxicity of ‘Soaps, stocks, vegetable oil, acidulated’. However, data is available for some of its constituents (see also table above):

·        Dietary exposure to 8.75% in diet (ca.4,375 mg/kg bw/day) glycerides with chain lengths of C8 - 18, including C18 -unsatd. was assessed in a two generation reproductive toxicity study in gerbils (Temmermans et al., 1988). No significant effect on frequency of litters, mean litter size, total no. of newborns and death during weanling stage were observed in the second generation. These results are in line with additional evidence of absence of developmental effects obtained from another two-generation study conducted in rats exposed to glycerides of chain length ranging from C16 -18, including C18 -unsatd (Nolen, 1972). Across all studies, tested doses ranged from 8.75 to 15% in diet. No significant toxicity was seen at any of the tested doses. The highest oral NOAEL could be considered to be 15% in diet, which is equivalent to an estimated 7,500 mg/kg bw/day.

·        No specific information could be found for fatty acids. 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 show any developmental toxicity.

·        Developmental toxicity studies conducted in animals with tocopherols revealed negative results (Fiume, 2002).

·        No developmental studies are available for sterols and sterols esters, although the detailed clinical examination of F1 and F2 litters from the 2-generation rat feeding study presented in Section 5.9.1. suggested slight potential for birth defects (ANZFA, 2001).

·        No specific information could be found for squalene. However, given its long history of safe use in nutritional and cosmetic applications, developmental toxicity is not expected.

Taken together, the above evidence suggests that ‘Soaps, stocks, vegetable oil, acidulated’ is not expected to be a developmental toxicant.

Justification for classification or non-classification

The above presented evidence suggests that ‘Soaps, stocks, vegetable oil, acidulated’ is not expected to be a reproductive or a developmental toxicant and therefore warrant any classification according to CLP Regulation EC/1272/2008.