Linseed oil, ester with pentaerythritol

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

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

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Principles of method if other than guideline:

A 13 week (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 days. 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 of each litter was autopsied. Liver and kidneys were weighed. For 5 males and 5 females selected randomly, these organs were examined microscopically.

GLP compliance:

no

Limit test:

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 days
- 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 days. 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:

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:

See section 7.5.1 for details on 13 WEEK FEEDING STUDY

REPRODUCTION SCREENING:
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 of each litter was autopsied. 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 days 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

Histopathological findings: non-neoplastic:

not examined

Other effects:

not examined

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 equivalent to 17 - 7 g/kg bw/day

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:

not specified

Clinical signs:

not specified

Key result

Critical effects observed:

not specified

Clinical signs:

not specified

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

Generation:

F1

Effect level:

> 15 other: % in diet equivalent to 7000 - 17000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects on maternal and pups

Key result

Critical effects observed:

not specified

Clinical signs:

not specified

Key result

Critical effects observed:

not specified

Key result

Reproductive effects observed:

not specified

None

Conclusions:

Under the study conditions, the NOAEL was considered to be 15% in diet, equivalent to 7000 - 17000 mg/kg bw/day, as no significant toxicity were observed on maternal rats and pups.

Executive summary:

A combined repeated and reproductive screening study was conducted to determine the effect of glycerides, C16-18 and C18-unsatd. (in the form of 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 days. 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 test substance did not show any adverse effects on male and female rats compared to other crude or heated vegetable oils. Furthermore, no signs of toxicity were observed on maternal rats or pups in the follow-up reproductive screening trial. Under the study conditions, the NOAEL was considered to be 15% in diet, equivalent to 7000 - 17000 mg/kg bw/day, as no significant toxicity were observed on maternal rats and pups (Coquet, 1977).

Reference 2

Endpoint:

three-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Principles of method if other than guideline:

Multigeneration breeding studies in rats were conducted according to the protocol used in earlier studies on unconventional oils (Rukmini, 1988 and 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

Limit test:

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 palmoilein
Group 4: 10% hydrogenated oil containing 30% of mahua oil

The test substances were mixed in the diet.

Details on mating procedure:

12 males and 12 females in each group were mated after 100-120 days. The mated animals were allowed to be together for a period of 18-20 days. 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 F. P. and Cerecodo L. R., 1948 A new method of reporting data on reproduction and lactation in the mouse. Scince 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, preweaning 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 weighted and the relative body weight were calculated.

Postmortem examinations (offspring):

Gross pathology and organ weight.

Statistics:

By using analyses of variance 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

Histopathological findings: non-neoplastic:

not examined

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

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: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects observed

Remarks on result:

not determinable

Remarks:

no NOAEL identified Generation: all generations

Clinical signs:

no effects observed

Reproductive function: oestrous cycle:

no effects observed

Key result

Dose descriptor:

NOAEL

Effect level:

10 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects observed

Key result

Critical effects observed:

not specified

Clinical signs:

effects observed, treatment-related

Mortality / viability:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings:

not examined

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 ration at birth and weaning did not indicate any difference in the relative fitness of each sex with maturity. Preweaning 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 generation, second mating, the group showed a comparatively high percentage, and in the second mating of the third generation, percentages were higher for all groups. These deaths may not be treatment related because the controls also had high preweaning mortality rates. Cannibalism was observed in some animals and the mortality of pups may be mainly due to this factor or to maternal neglect, 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 refelxological 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 F1b or Fb2 generations rats.

Gross pathology indicates no adverse abnormalities in the organs.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

10 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects observed

Remarks on result:

other:

Remarks:

no NOAEL identified Generation: all generations

Clinical signs:

no effects observed

Key result

Reproductive effects observed:

not specified

Conclusions:

Under the study condition, no adverse effect were shown on the reproductive or toxicological parameters studied at 10% substance in the diet.

Executive summary:

A three-generation study was conducted to determine the reproductive toxicity of glycerides, C16-18 and C18-unsatd. (in the form of red palm oil) using groups of 12 male and 12 female rats fed either groundnut oil (controls), red palm oil, refined, bleached and deodorized palmolein or hydrogenated vegetable oil containing mahua oil at 10% in the 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 measured between groups fed palm oil and controls. According to the authors, these results indicate that red palm oil can be safely used as an edible oil. Under the study condition, no adverse effect were shown on the reproductive or toxicological parameters studied at 10% substance in the diet (Manorama, 1993).

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

7 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Good quality studies

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity was assessed based on toxicity studies for substances representative of the main constituents of the test substance, namely glycerol esters (GE). The results are presented below:

Glycerol esters (GE)

Study 1:

A combined repeated and reproductive screening study was conducted to determine the effect of glycerides, C16-18 and C18-unsatd. (in the form of 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 days. 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 test substance did not show any adverse effects on male and female rats compared to other crude or heated vegetable oils. Furthermore, no signs of toxicity were observed on maternal rats or pups in the follow-up reproductive screening trial. Under the study conditions, the NOAEL was considered to be 15% in diet, equivalent to 7000 - 17000 mg/kg bw/day, as no significant toxicity were observed on maternal rats and pups (Coquet, 1977).

Study 2:

A three-generation study was conducted to determine the reproductive toxicity of glycerides, C16-18 and C18-unsatd. (in the form of red palm oil) using groups of 12 male and 12 female rats fed either groundnut oil (controls), red palm oil, refined, bleached and deodorized palmolein or hydrogenated vegetable oil containing mahua oil at 10% in the 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 measured between groups fed palm oil and controls. According to the authors, these results indicate that red palm oil can be safely used as an edible oil. Under the study condition, no adverse effect were shown on the reproductive or toxicological parameters studied at 10% substance in the diet (Manorama, 1993).

Effects on developmental toxicity

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

From October 03, 1994 to November 08, 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Lack of data on test substance

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

- lack of data on test substance

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:CD BR VAF/Plus

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Stone Ridge, NY, USA
- Age at study initiation: approximately 9 - 10 weeks
- Weight at study initiation: 200 - 274 g (female)
- Housing: The animals were housed in suspended stainless steel and wire mesh cages with absorbent paper below the cages. The females were housed separately during the study period except during mating.
- Diet: Purina Certified Rodent Chow No. 5002, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 13 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0 - 24.4
- Humidity (%): 40 - 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: October 03, 1994 to November 08, 1994

Route of administration:

oral: gavage

Vehicle:

other: polyethylene glycol (PEG 400)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The undiluted test substance was thoroughly mixed in vehicle prior to dispensing. The dosing solutions were prepared weekly.

VEHICLE
- Lot/batch no.: 122H1109
- Physical state: colorless liquid

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analysis for homogeneity of the test substance dilutions were performed on the lowest and the highest concentrations expected during the course of the study. The relative standard deviation ranged from 0.72 to 3.19%. Concentrations were analysed in the first and the third dosing mixture preparation. The analytical results for all test solutions were within 7% of the nominal concentrations for weeks 1 and 3.

Details on mating procedure:

- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug and/or sperm in vaginal smear referred to as day 0 of pregnancy
After confirmation of pregnancy, each mated female was returned to its cage and new females were placed into the males' cages until a required number of pregnant females were obtained.

Duration of treatment / exposure:

gestation days (GD) 6 - 15

Frequency of treatment:

daily, 7 d/week

Duration of test:

21 d (GD 0 - 21)

Dose / conc.:

100 mg/kg bw/day

Remarks:

Basis: actual ingested

Dose / conc.:

500 mg/kg bw/day

Remarks:

Basis: actual ingested

Dose / conc.:

1 000 mg/kg bw/day

Remarks:

Basis: actual ingested

No. of animals per sex per dose:

25 females per dose, 50 males in total

Control animals:

yes, concurrent vehicle

Details on study design:

- Rationale for animal assignment: Mated females were assigned to dose groups in the order of mating. Accordingly, the first confirmed mated female was assigned to Group 1, the next to Group 2 and so on until all mated animals for a given day were assigned to dose groups. On subsequent days, the next group in sequence was filled by the first confirmed mated female on that day and so on. Assignments were made until all groups were filled with confirmed mated females.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily during the treatment period and once daily at all other times during the study period

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: prior to selection and daily during gestation

BODY WEIGHT: Yes
- Time schedule for examinations: prior to selection and on GD 0, 6, 9, 12, 15, 18 and 21

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for examinations: concurrently with body weight examinations

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on GD 21
- all females were examined by gross necropsy

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter

Statistics:

Bartlett's test was performed to determine if the dose groups had equal variance (1% level of significance). If equal, the testing was done using parametric methods, otherwise nonparametric techniques were used. Parametric procedures: a standard one way ANOVA (F distribution) was used. If significant differences among the means were indicated, Dunnett's Test was used to determine which treatment groups differed significantly from control. In addition to the ANOVA, a standard regression analysis for linear response in the dose groups was performed that also tested for linear lack of fit in the model. Nonparametric procedures: the test of equality of means was performed using the Kruskal-Wallis Test. If significant differences among the means were indicated, Dunn's Summed Rank Test was used to determine which treatment groups differed significantly from the control. In addition, Jonckheere's Test for monotonic trend in the dose response was performed. All tests were conducted at the 5% and 1% level of significance. Fetal weight was analyzed by a standard nested analysis of covariance (fetuses nested within dams and dams nested within doses and litter size (both sexes combined)). If differences in groups were identified, the Least Significant Difference technique was used to determine which groups differed from the control group. Male and female fetuses were tested separately (the covariate was combined sexes in each analysis). Fetal malformation and variation incidence data were analyzed for statistical significance as follows: a standard chi-square analysis was performed to determine if the proportions of incidences differ between the groups tested. If anyone cell had an expected value less than 5, this step was not reported. Next, each treatment group was compared to the control group using a 2x2 Fisher Exact Test. Thirdly, Armitage's test for linear trend in the dosage groups was performed. All tests were reported at the 5% or 1% level of significance.

Indices:

Pre-implantation loss = (no. of Corpora Lutea - no. of Implantation Sites) / no. of Corpora Lutea * 100
Post-implantation loss = (no. of Implantation Sites - no. of live fetuses) / no. of Implantation Sites * 100

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Mortality:
No treatment-related mortality occurred. On GD 7, one female of the 500 mg/kg bw/d group was found dead. At necropsy, discolored and consolidated lungs were the only findings. The death of this female was probably a result of an accidental gavage error.

Clinical signs:
No treatment-related clinical signs were observed during the study period. The most frequently observation was soft stool, that occurred in treated and control animals and was therefore considered as a response to the vehicle rather than to the test substance. Other findings were scabs, little sign of stool, rales and alopecia in one or more groups. One female of the high dose group had a subcutis mass in the cervical area on GD 21. In summary, these clinical findings were considered to be incidental and unrelated to treatment with the test substance.

Body weight:
Treatment with the test substance had no statistically significant effect on mean body weight and mean body weight change at any interval.

Food consumption:
There were no statistically significant differences in mean food consumption between treated and control animals at any interval.

Post-mortem examinations:
There were no necropsy findings that were considered to be treatment-related. One high dose female had a subcutaneous mass in the cervical area and one control and one high dose female had dilated renal pelvis. In summary, these necropsy findings were considered to be incidental and unrelated to treatment.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: No adverse effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
Fetal body weight:
There were no statistically significant differences between treated and control mean fetal body weights of either sex (see Table 1).

Implantation data:
There were no statistically significant differences in the mean live fetuses, mean live male or female fetuses, mean resorptions, mean implantation sites, mean corpora lutea, mean total dead fetuses, mean fetuses per implantation sites, mean dead fetuses per implantation sites, mean resorptions per implantation sites, % pre-implantation loss, % post-implantation loss, mean malformed fetuses, mean fetuses with variations, or mean affected (resorptions + dead + malformed fetuses per litter) fetuses between treated and control groups. (see Table 1)

Fetal observations:
No biologically or statistically significant differences in total or individual variations or malformations were observed between the litters of test substance groups and control group (see Table 2). In the control, 100, 500 and 1000 mg/kg bw/d group, 5, 2, 6 and 1 fetuses, respectively, were stunted. External observations regarded as malformations were found in one fetus of the 100 mg/kg bw/d group (agnathia, astomia, low set ears, anophthalmia) and single findings of cleft palate, syndactyly and kinked tail in three fetuses of separate litters of the 500 mg/kg bw/d group. External variations were not observed in any group. Visceral observations regarded as malformations were limited to single occurrences of anophthalmia in one fetus each of the low and mid dose group, cleft palate and dilated brain ventricles in the 100 mg/kg bw/d group and two occurrences of folded retina in the control group. Visceral variations included a single or low incidence of dilated renal pelves, distended ureters and/or convoluted ureter. Skeletal observations regarded as malformations were limited to multiple skull bones malformed in one fetus of the 100 mg/kg bw/d group, short pubis in one fetus of the 1000 mg/kg bw/d group and one less presacral vertebrae in one fetus of the control, two fetuses in the low dose and one fetus in the high dose groups. Skeletal variations were observed throughout the groups and consisted primarily of hypoplastic, misshapen or unossified stemebrae, hypoplastic skull bones or pubis and rudimentary or misshapen ribs. In summary and without a clear pattern of response, all malformations were considered incidental and not to be treatment-related. Statistically significant differences in incidences of variations were limited to a decrease in hypoplastic skull bones of the mid dose group and a decrease in unossified stemebrae of the high dose group compared with controls. All variations were of no biological importance.

Skeletal ossification:
There were no statistically significant differences in mean ossification sites between treated and control groups.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed

Key result

Abnormalities:

not specified

Key result

Developmental effects observed:

not specified

Litter data and fetal body weight

	Control	dose groups [mg/kg bw/day]
		100	500	1000
No. of dams/litters examined	25	21	23	25
Mean no. of Corpora Lutea	16.8	16.8	17.7	16.8
	± 2.28	± 2.50	± 3.50	± 1.82
	n=25	n=20	n=23	n=25
Mean no. of Implantation Sites	15.76	15.9	16.09	15.52
	± 2.24	± 2.62	± 2.25	± 2.49
	n=25	n=21	n=23	n=25
Mean no. of resorptions	0.88	0.81	0.91	0.56
	± 1.05	± 1.29	± 1.12	± 0.82
	n=25	n=21	n=23	n=25
Pre-implantation loss [%]	5.7	6.7	8	7.7
	± 8.1	± 8.2	± 8.2	± 12.9
	n=25	n=20	n=23	n=25
Post-implantation loss per litter [%]	5.4	4.9	6	4.4
	± 6.4	± 7.7	± 7.3	± 6.4
	n=25	n=21	n=23	n=25
Mean no. of live fetuses	14.88	15.1	15.13	14.88
	± 2.19	± 2.64	± 2.46	± 2.64
	n=25	n=21	n=23	n=25
Mean no. of dead fetuses	0	0	0.04	0.08
			± 0.21	± 0.28
	n=25	n=21	n=23	n=25
Mean no. of female fetuses	7.72	7.48	8.04	7.8
	± 2.28	± 3.11	± 2.29	± 1.78
	n=25	n=21	n=23	n=25
Mean no. of male fetuses	7.16	7.62	7.09	7.08
	± 1.93	± 2.25	± 2.09	± 2.58
	n=25	n=21	n=23	n=25
Mean body weight (female fetuses) (g)	5.04	5.05	5.06	5.2
	± 0.44	± 0.32	± 0.46	± 0.37
	n=188	n=157	n=185	n=195
Mean body weight (male fetuses) (g)	5.33	5.27	5.39	5.44
	± 0.52	± 0.39	± 0.38	± 0.38
	n=175	n=160	n=163	n=177

Fetal variations and malformations

	Control	dose groups [mg/kg bw/day]
		100	500	1000
No. of fetuses with external variations	0	0	0	0
	n=372	n=317	n=347	n=358
No. of fetuses with external malformations	0	1	3	0
	n=372	n=317	n=347	n=358
No. of fetuses with visceral or head variations	0	0	1	2
	n=187	n=161	n=173	n=186/187*
No. of fetuses with visceral or head malformations	2	2	1	0
	n=187	n=161	n=173	n=186/187*
No. of fetuses with skeletal variations	45	39	29	35
	n=185	n=156	n=176	n=185
No. of fetuses with skeletal malformations	1	3	0	2
	n=185	n=156	n=176	n=185

*186 visceral examinations performed, 187 head examinations performed

Conclusions:

Under the study conditions, the NOAEL for embryo-/fetotoxicity and teratogenicity in rats was >1000 mg/kg bw/day.

Executive summary:

A study was conducted to determine developmental toxicity of fatty acids C8-10, mixed esters with dipentaerythritol, isooctanoic acid, pentaerythritol and tripentaerythritol according to OECD Guideline 414, in compliance with GLP. 50 male Sprague-Dawley rats were mated with females to achieve groups of 25 pregnant Sprague-Dawley rats which then received daily oral gavage doses of the test substance at concentrations of 100, 500 and 1000 mg/kg bw/day during Gestational Days 6 to 15. Control animals received the vehicle polyethylene glycol (PEG 400). On Day 21 of gestation, the animals were euthanized and examined for maternal and fetal parameters. There were no adverse effects found for all parameters examined in maternal animals. Based on the number of implantations, number of total litter losses by resorption, mortality, clinical signs, body weight, gross pathology and organ weights of maternal animals the NOAEL for maternal toxicity was found to be 1000 mg/kg bw/day. Examination of fetus litter size and weights, offspring viability (number alive and number dead), sex ratio, grossly visible abnormalities, external, head, soft tissue and skeletal abnormalities showed only incidental malformations in two high dose females. Under the study conditions, the NOAEL for embryo-/fetotoxicity and teratogenicity in rats was >1000 mg/kg bw/day (Exxon, 1995).

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

15 dams per group instead of 20, administration on day 0-19 of gestation, limited details on study design

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

(only 15 presumed pregnant females per group, exposure on day 0-19 of gestation, only 2 dose levels, nonstandard dermal exposure, limited details on exposure)

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Kingston, N.Y.
- Age at study initiation: approx. 9 weeks
- Mean weight at study initiation: 248 g
- Diet: Purina Certified Rodent Chow #5002 (Meal), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22
- Humidity (%): 40 - 60
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

dermal

Vehicle:

unchanged (no vehicle)

Details on exposure:

TEST SITE
- Type of wrap if used: open exposure, no wrap
- Time intervals for shavings or clippings: no data on frequency; clipped, intact skin
- Site: dorsal
Controls: The rats of the control group were clipped and collared. The intact dorsal skin of each rat was stroked with the tip of a syringe, but no test material was applied.

REMOVAL OF TEST SUBSTANCE
- Washing: no

TEST SUBSTANCE
- Amount(s) applied (volume or weight with unit): The amount of the test substance applied with a syringe was calculated based on the body weight of the animals and the density of the test substance.

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes. To minimize ingestion of the test material, the rats were fitted with cardboard Elizabethan-style collars.

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused: M/F ratio per cage: 1/1
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

gestation days 0 - 19

Frequency of treatment:

Daily

Duration of test:

The animals were sacrificed on day 20 of gestation.

Dose / conc.:

800 mg/kg bw/day (nominal)

Dose / conc.:

2 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

15 presumed-pregnant females

Control animals:

yes, concurrent no treatment

Details on study design:

- Dose selection rationale: based on results of a 13-week dermal study previously conducted with the same material

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once daily
- Cage side observations checked: signs of pathosis, abortion, premature delivery, and death

BODY WEIGHT: Yes
- Time schedule for examinations: day 0, 3, 6, 10, 13, 16, and 20 of gestation

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes
- Time schedule for calculation: days 0-3, 3-6, 6-10, 10-13, 13-16, and 16-20

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: The thoracic and abdominal cavities were exposed and all organs were examined grossly for evidence of pathosis.

OTHER:
- Clinical chemistry: alanine aminotransferase (ALT), albumin, albumin/globulin ration, alkaline phosphatase (ALP), aspartate aminotransferase (AST), bilirubin, calcium, chloride, cholesterol, creatinine, globulin, glucose, iron, lactate dehydrogenase (LDH), phosphorus, potassium, sodium, sorbitol dehydrogenase (SDH), total protein, triglycerides, urea nitrogen, and uric acid

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Number of live and dead fetuses: Yes
- Other: The ovaries of non-pregnant females were grossly examined and then discarded.

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: No

Statistics:

- Analysis of variances and group comparison using Fisher's Exact or Dunnett's test (maternal biophase and cesarean section data, and fetal data)
- ANOVA and Fisher's Exact test (fetal skeletal data)
- Fisher's Exact test (fetal visceral data)
- SAS procdures, Student-Newman-Keul's multiple comparison test (clinical chemistry data) P < 0.05

Details on maternal toxic effects:

Maternal toxic effects: yes. Remark: slight local effects

Details on maternal toxic effects:
- General observations: neck lesions, red nasal exudate, and chromodacryorrhea in all groups (considered not to be test substance-related as these signs are common in animals that are collared)
- Local effects: mild dermal irritation including erythema and flaking of the skin in the treatment groups
- Body weight: similar to controls in both treatment groups
- Body weight gain: similar to controls in both treatment groups
- Uterine and net body weights: similar to controls in both treatment groups
- Food consumption: similar to control in both treatment groups; only difference (statistically significant) in high dose group on day 13-16: 31.5 g vs. 29.5 g (corresponding control data)
- Necropsy: no remarkable findings were observed
- Fetal status and uterine position: no parameter evaluated appeared to be adversly affected
- Clinical chemistry: no differences between treated and non-treated rats were observed

Key result

Dose descriptor:

NOAEL

Effect level:

2 000 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: No adverse effects observed; slight local dermal irritation

Key result

Abnormalities:

not specified

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
- Fetal body measurements: Mean fetal body weights and crown-rump lengths, parameters of body growth and development were normal compared to controls.
- Fetal examination: No malformations or variations were observed. Bruises were observed on the skin of 4 fetuses from the control group and 2 fetuses form the 800 mg/kg bw/day group (considered to be incidental). One fetus from one dam exposed to 800 mg/kg bw/day was pale in colour. No other remarkable findings were observed during external examination.
- Malformations (lumbar and sacral vertebrae, tail, sternebrae, and ribs) were observed. The incidence was low and there was no apparent dose-response relationship, these effects were considered to be not treatment-related. Comparable incidences of variant skeletal development were observed in both control and treated fetuses.
- Visceral examinations: A statistically significant (high dose) increase in the number of fetuses with levocardia was observed. The response appeared to be dose-related
Levocardia:
Litters: control: 0 (0%); 800 mg/kg bw/day: 2 (14.3%); 2000 mg/kg bw/day: 7 (50%); 14 litters per group examined
Fetuses: control: 0 (0%); 800 mg/kg bw/day: 3 (3.2%); 2000 mg/kg bw/day: 7 (10.1%); 94, 93, and 99 fetuses examined, respectively
Microphthalmia, anophtalmia and "apparent" hydronephrosis were also observed. Variant visceral development was observed in control and treated fetuses.

Key result

Dose descriptor:

NOAEL

Remarks:

developmental

Effect level:

< 800 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Adverse effects observed

Key result

Dose descriptor:

LOAEL

Remarks:

developmental

Effect level:

800 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increased number of fetuses with levocardia

Key result

Abnormalities:

not specified

Key result

Developmental effects observed:

not specified

Levocardia was the only parameter affected in fetuses of dams treated with the test substance during gestation. In other studies, levocardia was observed in control fetuses, too. However, the effect of the test substance on heart development should be focused on in further studies as well as the impact, this effect has on postnatal survival.

Conclusions:

Under the study conditions, the NOAEL and LOAEL for embryo-/fetotoxicity and teratogenicity in rats for the test substance was found to be < 800 mg/kg bw/day and 800 mg/kg bw/day.

Executive summary:

A study was conducted to determine the developmental toxicity of fatty acids, C5-9, tetraesters with pentaerythritol according to OECD Guideline 414. Groups of 15 presumed pregnant female Sprague-Dawley rats received daily dermal doses of the test substance at concentrations of 800 and 2000 mg/kg bw/day during Gestational Days 0 to 19. Control animals remained untreated. On Day 20 of gestation, the animals were euthanized and examined for maternal and fetal parameters. There were no adverse effects found for all parameters examined in maternal animals. Based on the number of implantations, number of total litter losses by resorption, mortality, clinical signs, body weight, gross pathology and organ weights of maternal animals the NOAEL for maternal toxicity was found to be 2000 mg/kg bw/day. Examination of fetus litter size and weights, offspring viability (number alive and number dead), sex ratio, grossly visible abnormalities, external, head, soft tissue and skeletal abnormalities revealed no differences to controls and thus no indication for teratogenic effects. The only effect found was a dose-dependently increased number of fetuses with levocardia, although no hearth malformations have been detected. Furthermore levocardia was observed in vehicle control fetuses and in the control fetuses conducted in the test laboratory. Since levocardia was observed in both treated groups, the NOAEL for embryo-/fetotoxicity and teratogenicity in rats for test substance was found to be < 800 mg/kg bw/day and the LOAEL was 800 mg/kg bw/day (Exxon, 1988).

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Good quality studies

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Developmental toxicity was assessed based on toxicity studies for substances representative of the main constituents of the test substance, namely pentaerythritol esters (PE). The results are presented below:

 

Pentaerythritol esters (PE)

Study1:

A study was conducted to determine developmental toxicity of fatty acids C8-10, mixed esters with dipentaerythritol, isooctanoic acid, pentaerythritol and tripentaerythritol according to OECD Guideline 414, in compliance with GLP. 50 male Sprague-Dawley rats were mated with females to achieve groups of 25 pregnant Sprague-Dawley rats which then received daily oral gavage doses of the test substance at concentrations of 100, 500 and 1000 mg/kg bw/day during Gestational Days 6 to 15. Control animals received the vehicle polyethylene glycol (PEG 400). On Day 21 of gestation, the animals were euthanized and examined for maternal and fetal parameters. There were no adverse effects found for all parameters examined in maternal animals. Based on the number of implantations, number of total litter losses by resorption, mortality, clinical signs, body weight, gross pathology and organ weights of maternal animals the NOAEL for maternal toxicity was found to be 1000 mg/kg bw/day. Examination of fetus litter size and weights, offspring viability (number alive and number dead), sex ratio, grossly visible abnormalities, external, head, soft tissue and skeletal abnormalities showed only incidental malformations in two high dose females. Under the study conditions, the NOAEL for embryo-/fetotoxicity and teratogenicity in rats was >1000 mg/kg bw/day (Exxon, 1995).

Study 2:

A study was conducted to determine the developmental toxicity of fatty acids, C5-9, tetraesters with pentaerythritol according to OECD Guideline 414. Groups of 15 presumed pregnant female Sprague-Dawley rats received daily dermal doses of the test substance at concentrations of 800 and 2000 mg/kg bw/day during Gestational Days 0 to 19. Control animals remained untreated. On Day 20 of gestation, the animals were euthanized and examined for maternal and fetal parameters. There were no adverse effects found for all parameters examined in maternal animals. Based on the number of implantations, number of total litter losses by resorption, mortality, clinical signs, body weight, gross pathology and organ weights of maternal animals the NOAEL for maternal toxicity was found to be 2000 mg/kg bw/day. Examination of fetus litter size and weights, offspring viability (number alive and number dead), sex ratio, grossly visible abnormalities, external, head, soft tissue and skeletal abnormalities revealed no differences to controls and thus no indication for teratogenic effects. The only effect found was a dose-dependently increased number of fetuses with levocardia, although no hearth malformations have been detected. Furthermore levocardia was observed in vehicle control fetuses and in the control fetuses conducted in the test laboratory. Since levocardia was observed in both treated groups, the NOAEL for embryo-/fetotoxicity and teratogenicity in rats for test substance was found to be < 800 mg/kg bw/day and the LOAEL was 800 mg/kg bw/day (Exxon, 1988).

Justification for classification or non-classification

Based on the information available for substances representative of its main constituents, the test substance is not considered to meet the requirements for reproductive and developmental toxicity classification according to the EU CLP (Regulation 1272/2008/EC) criteria.