Rosin, maleated, esterified with sorbitol

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-03-04 to 2015-03-23

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

Study did not evaluate nor report data on fetal soft tissue, skeletal, or head effects.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

developmental toxicity

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2015-03-04 to 2015-03-23

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

Study did not evaluate nor report data on fetal soft tissue, skeletal, or head effects.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

Deviation had no impact on the integrity of the study

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Hydrocarbon Resins and Rosin Resins REACH Consortium (c/o Penman Consulting BVBA Rue Royale 157, Bte 13 B-1210 Brussels (Saint-Josse-ten-Noode), BELGIUM); Batch no: BG4J01332
- Expiration date of the lot/batch: 2015-10-21
- Purity test date: 2015-08-01

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored under nitrogen; used/formulated in air. Stored at -20°C; used/formulated at ambient temperature 10 to 30°C

FORM AS APPLIED IN THE TEST (if different from that of starting material): Yellow solid

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD®(SD) IGS BR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Age at study initiation: Not specified
- Weight at study initiation: 171 to 270 g
- Fasting period before study: not specified
- Housing: individually in solid-floor polypropylene cages with stainless steel mesh lids furnished with softwood flakes (Datesand Ltd., Cheshire, UK)
- Diet (e.g. ad libitum): Ground diet (Rodent PMI 5002 (Certified), BCM IPS Limited, London, UK) was used ad libitum
- Water (e.g. ad libitum): Mains drinking water was supplied ad libitum from polycarbonate bottles attached to the cage.
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3ºC
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least fifteen air changes per hour
- Photoperiod (hrs dark / hrs light): low intensity fluorescent lighting was controlled to give twelve hours continuous light and twelve hours darkness

IN-LIFE DATES: From: 2015-03-06 To: 2015-03-23

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): Dietary admixtures were prepared three times.
- Mixing appropriate amounts with (Type of food): A known amount of test item was mixed with a small amount of basal laboratory diet in a Robot Coupe Blixer 4 mixer until homogeneous. This pre-mix was then added to a larger amount of basal laboratory diet and mixed for a further thirty minutes at a constant speed, setting 1 in a Hobart QE200 mixer.
- Storage temperature of food: stored at room temperature

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. However, analysis of test item dietary formulations in a previous study (Harlan Study Number 41302579) demonstrated stability at 3000 and 20000 ppm for up to 8 days and confirmed the accuracyof the formulation procedure.

Details on mating procedure:

A total of thirty-two time-mated female Sprague-Dawley Crl:CD®(SD) IGS BR strain rats were obtained from Charles River (UK) Limited, Margate, Kent. Animals were delivered in one batch containing females prior to Day 3 of gestation. The day that positive evidence of mating was observed by the animal supplier was designated Day 0 of gestation.

Duration of treatment / exposure:

The test item was administered continuously, between days 3-19 of gestation, by dietary admixture.

Frequency of treatment:

Continuousl by dietary admixture

Duration of test:

Between days 3-19 of gestation

Dose / conc.:

0 ppm

Remarks:

Control

Dose / conc.:

3 000 ppm

Remarks:

Low Concentration (equivalent to a Mean Achieved Dose Level of 249.9 mg/kg bw/day)

Dose / conc.:

7 500 ppm

Remarks:

Intermediate Concentration (equivalent to a Mean Achieved Dose Level of 617.7 mg/kg bw/day)

Dose / conc.:

15 000 ppm

Remarks:

High Concentration (equivalent to a Mean Achieved Dose Level of 1198.5 mg/kg bw/day)

No. of animals per sex per dose:

8 per concentration

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: The dose levels were chosen based on previoustoxicity data and to approximate a highest achieved dosage of at least 1000 mg/kg bw/day. The oral route was selected as the most appropriate route of exposure, based on the physical properties of the test item, and the results of the study are believed to be of value in predicting the likely toxicity of the test item to man.
- Rationale for animal assignment (if not random): The animals were randomly allocated to treatment groups using a randomization procedure based on stratified body weight toensure similarity between the treatment groups. The animals were uniquely identified within the study by an ear punching system routinely used in these laboratories.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Following arrival, all animals were examined for overt signs of toxicity, ill-health or behavioral change once daily. All observations were recorded.

BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights were recorded on Days 3, 4, 5, 8, 11, 14, and 17 of gestation. Body weights were also recorded for animals at terminal kill (Day 20).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes; Food consumption was recorded for each individual animal for the periods Days 3 to 5, 5 to 8, 8 to 11, 11 to 14, 14 to 17 and 17 to 20.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water intake was observed daily by visual inspection of the water bottles for any overt changes.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: All animals were killed by carbon dioxide asphyxiation followed by cervical dislocation on Day 20 of gestation. All animals were subjected to a full external and internal examination and any macroscopic abnormalities were recorded. The ovaries and uteri of pregnant females were removed, examined and the following data recorded:

i) Number of corpora lutea
ii) Number, position and type of intrauterine implantation
iii) Fetal sex
iv) External fetal appearance
v) Fetal weight
vi) Placental weight
vii) Gravid uterus weight

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:
- Soft tissue examinations: No data
- Skeletal examinations: No data
- Head examinations: No data

Statistics:

Data were processed to give litter mean values, group mean values and standard deviations where appropriate, and were calculated to include data from all pregnant females with live fetuses on Day 20 of gestation. As the litter is the standard unit of assessment, individual litter values were first calculated and then group valuescalculated from these individual litter values.

Indices:

Pre and Post Implantation Loss
1) Percentage pre-implantation loss was calculated as: (number of corpora lutea - number of implantations/number of corpora lutea) x 100

2) Percentage post-implantation loss was calculated as: (number of implantations - number of live foetuses/number of implantations) x 100

Sex Ratio
Sex ratio was calculated for each litter value using the following formula:

% male fetuses (sex ratio) = (Number of male foetuses/Total number of foetuses) x 100

Historical control data:

Not specified

Clinical signs:

no effects observed

Description (incidence and severity):

No clinical signs were observed on the study.

Mortality:

no mortality observed

Description (incidence):

There were no unscheduled deaths.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

At 15000 ppm, body weight gain between Days 3-4, Days 11-14 and Days 17-20 was slightly lower than control. However, overall body weight gain during gestation, when adjusted for the contribution of the gravid uterus was comparable to control.

At 7500 and 3000 ppm overall body weight gain during gestation was considered to be unaffected by treatment.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

There was no clear adverse effect on food consumption during gestation at 3000, 7500 and 15000 ppm.

At 15000 ppm, food consumption was slightly reduced between Days 3-5, however, differences from controls were not marked and recovery was evident thereafter.

Water consumption and compound intake (if drinking water study):

no effects observed

Description (incidence and severity):

Daily visual inspection of water bottles did not reveal any overt intergroup differences.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No abnormalities were detected during the macroscopic examination of the pregnant females at termination on Day 20 of gestation.

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on litterdata as assessed by numbers of implantations.

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on in-utero offspring survival (as assessed by the mean numbers of early or late resorptions).

Early or late resorptions:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on in-utero offspring survival (as assessed by the mean numbers of early or late resorptions).

Dead fetuses:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on litter data as assessed by live litter size.

Other effects:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on mean fetal or placental weight at 3000, 7500 or 15000 ppm.

Details on maternal toxic effects:

There was no effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio and pre and post-implantation losses at 3000, 7500 or 15000 ppm. There was no effect of maternal treatment on mean fetal or placental weight at 3000, 7500 or 15000 ppm. A slight reduction in litter weight and total placental weight was evident at 15000 ppm however this may have been the result of one female (No. 31) which had a smaller litter size than the remaining females in this group and in the control group.

Dose descriptor:

NOAEL

Effect level:

15 000 ppm

Based on:

test mat.

Basis for effect level:

other: Systemic Toxicity

Abnormalities:

no effects observed

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): There was no effect of maternal treatment on mean fetal at 3000, 7500 or 15000 ppm.

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

effects observed, non-treatment-related

Description (incidence and severity):

A slight reduction in litter weight was evident at 15000 ppm however this may have been the result of one female (No. 31) which had a smaller litter size than the remaining females in this group and in the control group.

External malformations:

no effects observed

Description (incidence and severity):

There were no findings apparent for fetuses from treated females at external examination on Day 20 of gestation.

Dose descriptor:

NOAEL

Effect level:

15 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Developmental Toxicity

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

The systemic and developmental toxicity NOAEL for RARA, fumarated esters with glycerol in rats is therefore determined to be 15000 ppm (equivalent to 1198.5 mg/kg bw/day).

Overall, there were no findings observed in this preliminary study that were considered to exclude the use of 15000 ppm as the high dosage for any further investigation of embryofoetal toxicity in the rat. Dosing from Day 3 of gestation using dosages of 0 (Control), 3000, 7500 and 15000 ppm is therefore recommended for the planned rat pre-natal development toxicity study.

Executive summary:

In a key pre-natal developmental toxicity study, the test material (RARA, fumarated, esters with glycerol (CAS# 97489-11-7)), was administered by continuous dietary admixture to three groups each of eight time-mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between Days 3 and 19 of gestation inclusive at dietary concentrations of 3000, 7500, and 15000 ppm (equivalent to a mean achieved dosage of 249.9, 617.7 and 1198.5 mg/kg bw/day respectively). A further group of eight time mated females was treated with basal laboratory diet to serve as a control.

 

Clinical signs, body weight change, food and water consumptions were monitored during the study. All females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weights, fetal weight, sex and external macroscopic appearance of fetuses were recorded.

 

There were no unscheduled deaths or signs of test material-related clinical signs of toxicity observed through the study period. At 15000 ppm, body weight gain between Days 3-4, Days 11-14 and Days 17-20 was slightly lower than control. However, overall body weight gain during gestation, when adjusted for the contribution of the gravid uterus was comparable to control. At 7500 and 3000 ppm, overall body weight gain during gestation was considered to be unaffected by treatment. Food and water consumption remained unaffected by treatment at 3000, 7500 and 15000 ppm and gross necropsy did not reveal any remarkable findings.

 

There was no effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio and pre-and post-implantation losses or on mean fetal, litter or placental weights at 3000, 7500 or 15000 ppm. There were no findings apparent for fetuses from treated females at external examination on Day 20 of gestation.

 

The systemic and developmental toxicity NOAEL for RARA, fumarated esters with glycerol in rats is therefore determined to be 15000 ppm (equivalent to 1198.5 mg/kg bw/day).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

Deviation had no impact on the integrity of the study

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Resin acids and Rosin acids, fumarated, esters with glycerol

EC Number:

307-051-0

EC Name:

Resin acids and Rosin acids, fumarated, esters with glycerol

Cas Number:

97489-11-7

Details on test material:

- Name of test material (as cited in study report): 84-0981; UNI-REZ 757
- Molecular formula (if other than submission substance):
- Substance type: Reaction product derived from UVCB
-

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Hydrocarbon Resins and Rosin Resins REACH Consortium (c/o Penman Consulting BVBA Rue Royale 157, Bte 13 B-1210 Brussels (Saint-Josse-ten-Noode), BELGIUM); Batch no: BG4J01332
- Expiration date of the lot/batch: 2015-10-21
- Purity test date: 2015-08-01

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Stored under nitrogen; used/formulated in air. Stored at -20°C; used/formulated at ambient temperature 10 to 30°C

FORM AS APPLIED IN THE TEST (if different from that of starting material): Yellow solid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD®(SD) IGS BR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Age at study initiation: Not specified
- Weight at study initiation: 171 to 270 g
- Fasting period before study: not specified
- Housing: individually in solid-floor polypropylene cages with stainless steel mesh lids furnished with softwood flakes (Datesand Ltd., Cheshire, UK)
- Diet (e.g. ad libitum): Ground diet (Rodent PMI 5002 (Certified), BCM IPS Limited, London, UK) was used ad libitum
- Water (e.g. ad libitum): Mains drinking water was supplied ad libitum from polycarbonate bottles attached to the cage.
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3ºC
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least fifteen air changes per hour
- Photoperiod (hrs dark / hrs light): low intensity fluorescent lighting was controlled to give twelve hours continuous light and twelve hours darkness

IN-LIFE DATES: From: 2015-03-06 To: 2015-03-23

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): Dietary admixtures were prepared three times.
- Mixing appropriate amounts with (Type of food): A known amount of test item was mixed with a small amount of basal laboratory diet in a Robot Coupe Blixer 4 mixer until homogeneous. This pre-mix was then added to a larger amount of basal laboratory diet and mixed for a further thirty minutes at a constant speed, setting 1 in a Hobart QE200 mixer.
- Storage temperature of food: stored at room temperature

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. However, analysis of test item dietary formulations in a previous study (Harlan Study Number 41302579) demonstrated stability at 3000 and 20000 ppm for up to 8 days and confirmed the accuracyof the formulation procedure.

Details on mating procedure:

A total of thirty-two time-mated female Sprague-Dawley Crl:CD®(SD) IGS BR strain rats were obtained from Charles River (UK) Limited, Margate, Kent. Animals were delivered in one batch containing females prior to Day 3 of gestation. The day that positive evidence of mating was observed by the animal supplier was designated Day 0 of gestation.

Duration of treatment / exposure:

The test item was administered continuously, between days 3-19 of gestation, by dietary admixture.

Frequency of treatment:

Continuousl by dietary admixture

Duration of test:

Between days 3-19 of gestation

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 ppm

Remarks:

Control

Doses / concentrations 2

Dose / conc.:

3 000 ppm

Remarks:

Low Concentration (equivalent to a Mean Achieved Dose Level of 249.9 mg/kg bw/day)

Doses / concentrations 3

Dose / conc.:

7 500 ppm

Remarks:

Intermediate Concentration (equivalent to a Mean Achieved Dose Level of 617.7 mg/kg bw/day)

Doses / concentrations 4

Dose / conc.:

15 000 ppm

Remarks:

High Concentration (equivalent to a Mean Achieved Dose Level of 1198.5 mg/kg bw/day)

No. of animals per sex per dose:

8 per concentration

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: The dose levels were chosen based on previoustoxicity data and to approximate a highest achieved dosage of at least 1000 mg/kg bw/day. The oral route was selected as the most appropriate route of exposure, based on the physical properties of the test item, and the results of the study are believed to be of value in predicting the likely toxicity of the test item to man.
- Rationale for animal assignment (if not random): The animals were randomly allocated to treatment groups using a randomization procedure based on stratified body weight toensure similarity between the treatment groups. The animals were uniquely identified within the study by an ear punching system routinely used in these laboratories.

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Following arrival, all animals were examined for overt signs of toxicity, ill-health or behavioral change once daily. All observations were recorded.

BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights were recorded on Days 3, 4, 5, 8, 11, 14, and 17 of gestation. Body weights were also recorded for animals at terminal kill (Day 20).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes; Food consumption was recorded for each individual animal for the periods Days 3 to 5, 5 to 8, 8 to 11, 11 to 14, 14 to 17 and 17 to 20.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water intake was observed daily by visual inspection of the water bottles for any overt changes.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: All animals were killed by carbon dioxide asphyxiation followed by cervical dislocation on Day 20 of gestation. All animals were subjected to a full external and internal examination and any macroscopic abnormalities were recorded. The ovaries and uteri of pregnant females were removed, examined and the following data recorded:

i) Number of corpora lutea
ii) Number, position and type of intrauterine implantation
iii) Fetal sex
iv) External fetal appearance
v) Fetal weight
vi) Placental weight
vii) Gravid uterus weight

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:
- Soft tissue examinations: No data
- Skeletal examinations: No data
- Head examinations: No data

Statistics:

Data were processed to give litter mean values, group mean values and standard deviations where appropriate, and were calculated to include data from all pregnant females with live fetuses on Day 20 of gestation. As the litter is the standard unit of assessment, individual litter values were first calculated and then group valuescalculated from these individual litter values.

Indices:

Pre and Post Implantation Loss
1) Percentage pre-implantation loss was calculated as: (number of corpora lutea - number of implantations/number of corpora lutea) x 100

2) Percentage post-implantation loss was calculated as: (number of implantations - number of live foetuses/number of implantations) x 100

Sex Ratio
Sex ratio was calculated for each litter value using the following formula:

% male fetuses (sex ratio) = (Number of male foetuses/Total number of foetuses) x 100

Historical control data:

Not specified

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

no effects observed

Description (incidence and severity):

No clinical signs were observed on the study.

Mortality:

no mortality observed

Description (incidence):

There were no unscheduled deaths.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

At 15000 ppm, body weight gain between Days 3-4, Days 11-14 and Days 17-20 was slightly lower than control. However, overall body weight gain during gestation, when adjusted for the contribution of the gravid uterus was comparable to control.

At 7500 and 3000 ppm overall body weight gain during gestation was considered to be unaffected by treatment.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

There was no clear adverse effect on food consumption during gestation at 3000, 7500 and 15000 ppm.

At 15000 ppm, food consumption was slightly reduced between Days 3-5, however, differences from controls were not marked and recovery was evident thereafter.

Water consumption and compound intake (if drinking water study):

no effects observed

Description (incidence and severity):

Daily visual inspection of water bottles did not reveal any overt intergroup differences.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No abnormalities were detected during the macroscopic examination of the pregnant females at termination on Day 20 of gestation.

Maternal developmental toxicity

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on litterdata as assessed by numbers of implantations.

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on in-utero offspring survival (as assessed by the mean numbers of early or late resorptions).

Early or late resorptions:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on in-utero offspring survival (as assessed by the mean numbers of early or late resorptions).

Dead fetuses:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on litter data as assessed by live litter size.

Other effects:

no effects observed

Description (incidence and severity):

There was no effect of maternal treatment on mean fetal or placental weight at 3000, 7500 or 15000 ppm.

Details on maternal toxic effects:

There was no effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio and pre and post-implantation losses at 3000, 7500 or 15000 ppm. There was no effect of maternal treatment on mean fetal or placental weight at 3000, 7500 or 15000 ppm. A slight reduction in litter weight and total placental weight was evident at 15000 ppm however this may have been the result of one female (No. 31) which had a smaller litter size than the remaining females in this group and in the control group.

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

15 000 ppm

Based on:

test mat.

Basis for effect level:

other: Systemic Toxicity

Maternal abnormalities

Abnormalities:

no effects observed

Results (fetuses)

Description (incidence and severity):

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): There was no effect of maternal treatment on mean fetal at 3000, 7500 or 15000 ppm.

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

effects observed, non-treatment-related

Description (incidence and severity):

A slight reduction in litter weight was evident at 15000 ppm however this may have been the result of one female (No. 31) which had a smaller litter size than the remaining females in this group and in the control group.

External malformations:

no effects observed

Description (incidence and severity):

There were no findings apparent for fetuses from treated females at external examination on Day 20 of gestation.

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

15 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Developmental Toxicity

Fetal abnormalities

Abnormalities:

no effects observed

Overall developmental toxicity

Developmental effects observed:

no

Applicant's summary and conclusion

Conclusions:

The systemic and developmental toxicity NOAEL for RARA, fumarated esters with glycerol in rats is therefore determined to be 15000 ppm (equivalent to 1198.5 mg/kg bw/day).

Overall, there were no findings observed in this preliminary study that were considered to exclude the use of 15000 ppm as the high dosage for any further investigation of embryofoetal toxicity in the rat. Dosing from Day 3 of gestation using dosages of 0 (Control), 3000, 7500 and 15000 ppm is therefore recommended for the planned rat pre-natal development toxicity study.

Executive summary:

In a key pre-natal developmental toxicity study, the test material (RARA, fumarated, esters with glycerol (CAS# 97489-11-7)), was administered by continuous dietary admixture to three groups each of eight time-mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between Days 3 and 19 of gestation inclusive at dietary concentrations of 3000, 7500, and 15000 ppm (equivalent to a mean achieved dosage of 249.9, 617.7 and 1198.5 mg/kg bw/day respectively). A further group of eight time mated females was treated with basal laboratory diet to serve as a control.

 

Clinical signs, body weight change, food and water consumptions were monitored during the study. All females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weights, fetal weight, sex and external macroscopic appearance of fetuses were recorded.

 

There were no unscheduled deaths or signs of test material-related clinical signs of toxicity observed through the study period. At 15000 ppm, body weight gain between Days 3-4, Days 11-14 and Days 17-20 was slightly lower than control. However, overall body weight gain during gestation, when adjusted for the contribution of the gravid uterus was comparable to control. At 7500 and 3000 ppm, overall body weight gain during gestation was considered to be unaffected by treatment. Food and water consumption remained unaffected by treatment at 3000, 7500 and 15000 ppm and gross necropsy did not reveal any remarkable findings.

 

There was no effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio and pre-and post-implantation losses or on mean fetal, litter or placental weights at 3000, 7500 or 15000 ppm. There were no findings apparent for fetuses from treated females at external examination on Day 20 of gestation.

 

The systemic and developmental toxicity NOAEL for RARA, fumarated esters with glycerol in rats is therefore determined to be 15000 ppm (equivalent to 1198.5 mg/kg bw/day).