Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Oral (OECD 422): NOAEL (Parental general toxicity) = 1000 mg/kg bw/day , NOEL (reproduction/developmental) = 1000 mg/kg bw/day 

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-06-28 to 2013-03-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted under GLP according to OECD TG 422.
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: OPPTS 870.3650 (Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories BV, Netherlands
- Age at study initiation: 11 weeks
- Weight at study initiation: 290 to 326 g (males), 162 to 222 g (females)
- Fasting period before study: no
- Housing: in groups of three to five animals in Makrolon type-4 cages with wire mesh tops up to the day of randomization and afterwards individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Storage temperature of food: at room temperature (20 ± 5 °C) in glass beakers

VEHICLE
- Justification for use and choice of vehicle (if other than water): no data
- Concentration in vehicle: 0, 12.5, 75, 250 mg/mL
- Amount of vehicle: 4 mL/kg bw
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 14 days maximum
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
On the first treatment day samples from the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. To confirm the stability (8 days) samples of about 2 g of each concentration were taken from the middle of each aliquot used on day 7 of the treatment. During the last week of the treatment, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and delivered on dry ice to analytic laboratory and stored there at -20 ± 5 °C until analysis. The samples received were dissolved in TBME by sonication for at least 5 minutes and then diluted to volume with TBME. Sample solutions were further diluted with TBME into the calibration range. The samples were analysed with gas chromatography coupled to an FID.
Duration of treatment / exposure:
The test substance was administered to male rats for 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 3 post-partum, one day before necropsy.
Frequency of treatment:
daily
Details on study schedule:
not relevant
Dose / conc.:
50 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
11
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on a previous non-GLP dose range-finding toxicity study in Han Wistar rats, using dose levels of 0, 50, 200 and 1000 mg/kg/day.
Positive control:
no data
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily cage-side clinical observations (once daily, during acclimatization and up to day of necropsy). Additionally females were observed for signs of difficult or prolonged parturition, and behavioural abnormalities in nesting and nursing.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: In males, it was performed once prior to the first administration of the test item and weekly thereafter. In females, it was prepared once prior to the first administration of the test item, weekly during the pre-pairing and pairing periods and on days 0, 6, 13 and 20 of the gestation period.

BODY WEIGHT: Yes
- Time schedule for examinations: daily from treatment start to day of necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Males: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 13; after pairing period weekly.
- Females: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 13; gestation days 0 - 7, 7 - 14 and 14 - 21 and days 1 - 4 of the lactation.

OTHER: HEMATOLOGY, CLINICAL CHEMISTRY, NEUROBEHAVIOURAL EXAMINATION
Oestrous cyclicity (parental animals):
Not examined
Sperm parameters (parental animals):
Not examined
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring: number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies

GROSS EXAMINATION OF DEAD PUPS:
Yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals were sacrificed after treatment for 28 days
- Maternal animals: All surviving animals were sacrificed on day 4 post-partum

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, abdominal viscera and organs of the reproductive system

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in attached document [Organ Weights and Preservation] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 4 days of age.
- These animals were subjected to post-mortem examinations (macroscopic and/or microscopic examination) as follows: no data

GROSS NECROPSY
no data

HISTOPATHOLOGY / ORGAN WEIGTHS
no data
Statistics:
The following statistical methods were used to analyze food consumption, body weights and reproduction data:
• Means and standard deviations of various data were calculated.
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test was applied if the variables could be dichotomized without loss of information.
Reproductive indices:
From the on-line recorded reproduction data, the following parameters were calculated: mean precoital time, percentage mating, fertility index, conception rate, post-implantation loss, gestation index, birth index and viability index.
Offspring viability indices:
From the on-line reproduction data, the following parameters were calculated: dead/live pups at first litter check, pup sex ratio and viability index.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
please see Details on results (parentals animals)
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effects on absolute body weight or body weight gain were noted in males and females at any dose level.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No effects on absolute body weight or body weight gain were noted in males and females at any dose level.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
please see Details on results (parent animals)
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
During macroscopical examination enlarged liver was found in one female at the dose level of 1000 mg/kg bw/day. This finding was considered to be related to the treatment with the test item.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
please see Details on results (parent animals)
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Test substance intake: gavage
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY
At the dose level of 1000 mg/kg bw/day, bedding in mouth and salivation in males starting from day 6 of the pairing period and in females starting from day 1 of the pairing period. At the dose level of 300 mg/kg bw/day, bedding in mouth in females starting from day 1 of the gestation period was observed. These observations were considered to be sign of discomfort caused by the treatment with the test item.

BODY WEIGHT AND FOOD CONSUMPTION
No effects on absolute body weight or body weight gain were noted in males and females at any dose level.

TEST SUBSTANCE INTAKE
No effects (gavage) observed.

REPRODUCTIVE PERFORMANCE
Mating performance, fertility, corpora lutea count, duration of gestation, implantation rate and post-implantation loss, litter size or postnatal loss were not affected by the treatment with the test item at any dose level.

ORGAN WEIGHTS
At the dose level of 1000 mg/kg bw/day, statistically significantly increased liver weights were noted in males. This finding was considered to be due to adaptive changes in the liver metabolism caused by the treatment with the test item and not adverse.

GROSS PATHOLOGY
During macroscopically examination enlarged liver was found in one female at the dose level of 1000 mg/kg bw/day. This finding was considered to be related to the treatment with the test item.

HISTOPATHOLOGY
Microscopic changes related to the treatment with the test item were observed in the liver of males and females at the dose level of 1000 mg/kg bw/day and in the kidneys of males at the dose levels of 1000 and 300 mg/kg bw/day.

Liver and thyroid gland
Centrilobular hypertrophy of the hepatocytes was seen in males and females at the dose level of 1000 mg/kg bw/day with a higher incidence and severity present in males. In males at the dose level of 1000 mg/kg bw/day, hypertrophy of the follicular epithelium of the thyroid gland was also noted. This finding was considered to be very likely secondary to the increased metabolism of T4 and T3 in the liver that in turn stimulated the secretion of TSH from the pituitary and subsequent follicular epithelial hypertrophy of the thyroid.

Kidneys
At the dose level of 1000 and 300 mg/kg bw/day, hyaline droplets accumulation was present in the proximal convoluted tubules in the kidneys of males, which was associated to an increased incidence of tubular basophilia and to the presence of granular casts in one male at the high-dose level. In one female at the dose level of 1000 mg/kg bw/day (no. 88), an unilateral nephroblastoma was observed in the kidney and considered to be of spontaneous occurrence and therefore unrelated to the treatment with the test item. The remainder of findings noted was within the range of normal background lesions which may be recorded in rats of this strain and

OTHER FINDINGS
The examination of the ovaries and the feminine genital tract did not reveal any difference in estrus cycling between control and treated animals. The qualitative assessment of the male genital organs did not reveal any treatment-related effect.
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: The highest applied dose
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: The highest applied dose
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
not specified
Other effects:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
VIABILITY
No effects observed.

CLINICAL SIGNS
No effects observed.

BODY WEIGHT
No effects observed.

SEXUAL MATURATION
No effects observed.

ORGAN WEIGHTS
No effects observed.

GROSS PATHOLOGY
No effects observed.
Dose descriptor:
NOEL
Remarks:
reproductive/developmental
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: The highest applied dose
Reproductive effects observed:
not specified
Conclusions:
Based on the study findings in the rats, the relevant NOAEL for human toxicity is 1000 mg/kg bw/day. The NOEL for reproduction/developmental toxicity is considered to be 1000 mg/kg bw/day.
Executive summary:

The purpose of this study was to generate preliminary information concerning the effects of the substance on the possible health hazards likely to arise from repeated exposure over a relatively limited period of time. In addition it provides information on possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition. The substance was administered to male rats for 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 3 post-partum, one day before necropsy.

The following dose levels were applied:

Group 1: 0 mg/kg body weight/day (control group)

Group 2: 50 mg/kg body weight/day

Group 3: 300 mg/kg body weight/day

Group 4: 1000 mg/kg body weight/day

A standard dose volume of 4 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (corn oil).

Parent Animals

Mortality and General Tolerability

All animals survived scheduled study period. Bedding in mouth and salivation were noted in males and females at the dose level of 1000 mg/kg bw/day. Bedding in mouth alone was noted in females at the dose level of 300 mg/kg bw/day. These observations were considered to be sign of discomfort caused by the treatment with the test item. No further clinical signs or observations were noted in males or females at any dose level.

Functional Observational Battery

With exception for salivation noted in one male and one female at the dose level of 1000 mg/kg bw/day, no further test item-related findings were noted during the tests of functional observational battery or measurement of locomotor activity.

Food Consumption

At the dose level of 1000 mg/kg bw/day in females, transient reduction in food consumption was noted at the beginning of the treatment. No test item-related effects on food consumption were noted in males at any dose level or females at dose levels of 50 and 300 mg/kg bw/day.

Body Weights

No effects on absolute body weights or body weight gain were noted in males or females at any dose level.

Clinical Laboratory Investigations

In males statistically significantly higher creatinine concentration was noted in all dose groups. This finding was accompanied by statistically significantly reduced concentration of phosphorus in high- and mid-dose groups. These findings were considered to most probably be secondary to lesions in the kidneys which were considered sex- and species-specific for male rats and therefore not relevant for humans. Further findings were statistically significantly lower concentrations of total bilirubin and statistically significantly lower concentration of bile acids in males at the dose level of 1000 mg/kg bw/day. These findings were considered to possibly be secondary to the adaptive changes in the liver metabolism caused by the treatment with the test item and not to be adverse. No further test item-related changes in hematology or biochemistry parameters were noted in males or females at any dose level.

Reproduction and Breeding Data

Mating performance, fertility, corpora lutea count, duration of gestation, implantation rate and post-implantation loss, litter size or postnatal loss were not affected by the treatment with the test item at any dose level.

Organ Weights

At the dose level of 1000 mg/kg bw/day, statistically significantly increased liver weights were noted in males. This finding was considered to be due to adaptive changes in the liver metabolism caused by the treatment with the test item and not adverse.

Macroscopical Findings and Histopathological Examinations

During macroscopical examination enlarged liver was found in one female at the dose level of 1000 mg/kg bw/day. This finding was considered to be related to the treatment with the test item.

During microscopic examinations, test item-related findings were noted in the liver of males and females at the dose level of 1000 mg/kg bw/day. These changes consisted of centrilobular hypertrophy of the hepatocytes and were considered to be an adaptive change due to the increased metabolism of the liver. In addition, hypertrophy of the follicular epithelium of the thyroid gland was noted in males at the dose level of 1000 mg/kg bw/day. This finding was considered to be very likely secondary to the increased metabolism in the liver. Further findings related to the treatment were noted in the kidneys of males at the dose levels of 1000 and 300 mg/kg bw/day; hyaline droplets associated with tubular basophilia and presence of granular casts. These findings were considered to be specific for male rats and adverse in this species but not toxicologically relevant for humans.

Litter Data - F1 Pups

Findings at First Litter Check and during Lactation

No test item-related findings were noted in pups during first litter check and during lactation at any dose level. Pups sex ratio was not affected by the exposure to the test item at any dose level.

Pup Weights to Day 4 Post-Partum

Body Weights and body weight gain of pups were not affected by the treatment with the test item at any dose level.

Macroscopical Findings

No macroscopical findings were noted in pups at any dose level.

Conclusion

Based on the lesions in the kidneys found in males at the dose levels of 1000 and 300 mg/kg bw/day which were considered to be adverse, general NOAEL (No Observed Adverse Effect Level) for males was established at 50 mg/kg bw/day. However, lesions in the kidneys were considered to be specific for male rats and therefore not toxicologically relevant for humans. NOAEL for females was considered to be 1000 mg/kg bw/day, the highest dose level used in the study. The NOEL for reproduction/developmental toxicity was considered to be 1000 mg/kg bw/day, the highest dose level used in the study.

Effect on fertility: 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:
The study is GLP according to OECD TG 422.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

- Reproduction toxicity:

In reproduction/developmental screening study with the test substance (2013), the NOEL for reproductive/developmental effects was determined at 1000 mg/kg bw/day. Since the NOAEL for parental general toxicity was established at 1000 mg/kg bw/day (the highest applied dose), it could be concluded that the substance does not need to be classified as reproductive toxicant.

Effects on developmental toxicity

Description of key information

Oral (OECD 414): NOAEL (maternal/embryo-/fetotoxicity/teratogenicity) ≥1000 mg/kg bw/day, NOEL (maternal toxicity) = 300 mg/kg bw/day

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May-October 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted under GLP according to OECD TG 414.
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Canada Inc., Quebec, Canada
- Age at study initiation: 11 - 12 weeks
- Weight at study initiation: 242 - 333 g
- Fasting period before study: no
- Housing: animals were housed individually in stainless steel cages.
- Diet: standard certified commercial chow (ENVIGO Certified Global Rodent Diet #2018C), ad libitum
- Water: municipal tap water, ad libitum
- Acclimation period: at least 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 50 ± 20
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS
The test item and reference item dosing formulations were prepared at least weekly. The test item was weighed and transferred into a container (or weighed directly into the container). The vehicle was then added to the final volume, followed by stirring. All formulations, including the vehicle, were stored at room temperature prior to administration.

VEHICLE
- Justification for use and choice of vehicle: The vehicle was chosen due to its ability to solubilize the test item.
- Concentration in vehicle: 25, 75 and 250 mg/mL
- Amount of vehicle: 4 mL/kg bw
- Lot/batch no.: MKBV2080V
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
To verify the concentration and homogeneity of the test item in the formulations, representative samples (5 mL in duplicate) were taken from the top, middle, and bottom of each concentration at the start of the study, and from the middle of a preparation conducted towards the end of the treatment period. The samples were stored frozen (-20 °C). An additional 15-mL sample was collected from the middle of each concentration at the start of the study for density measurement. Following confirmation of safe and intact receipt of the first set of samples, the second set of the duplicates were discarded. The analysis was performed by the analytical laboratory by gas chromatography (GC analysis) within the expected 1-month stability data range. An acceptance criterion for the formulation was ± 15% of nominal.
Details on mating procedure:
- Animals had been mated at the animal supplier on the day prior to shipping, and on the following day, those females confirmed as mated (with vaginal plugs) were shipped to the testing laboratory.
- Proof of pregnancy: vaginal plug, referred to as day 0 of pregnancy
Duration of treatment / exposure:
Day 5 - 20 of gestation
Frequency of treatment:
once daily, 7 days/week
Duration of test:
16 days; Day 5 - 20 of gestation
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Remarks:
based on oral range finding study
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Remarks:
based on oral range finding study
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Remarks:
based on oral range finding study
No. of animals per sex per dose:
23 P females (100 mg/kg bw/day)
24 P females (0, 300 and 1000 mg/kg bw/day)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on an oral range finding gavage study conducted in rats with the test item, the high dose level selected by the Sponsor for this dose range finding study is 1000 mg/kg bw/day, with approximately 3-fold lower Mid and Low dose levels (300 and 100 mg/kg, respectively).
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Clinical signs (ill health, behavioral changes, etc.) and mortality were recorded on all animals twice daily throughout the course of the study. Additional clinical observations may be performed if deemed necessary to properly monitor the animals health condition.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: A detailed clinical examination was performed on each animal on Gestation Days 0, 3, 5, 6, 9, 12, 15, 18 and 21 (day of necropsy).

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded for all animals at least as follows: Gestation Days 0, 3, 5, 6, 9, 12, 15, 18 and 21 (prior to Caesarean section).

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Individual food intake was measured on all surviving animals at least as follows: Gestation Days 0-3, 3-5, 5-6, 6-9, 9-12, 12-15, 15-18 and 18-21.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on Gestation Day 21
- Organs examined: Reproductive tract including ovaries and gravid uterus with placenta (external macroscopic examination, including identification of all clinically-recorded lesions) as well as a detailed internal examination will be performed.
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:
Each pairwise group comparisons of interest were conducted via a two-sided test at the 5% significance level (with significant results p ≤0.001, p ≤0.01 or p ≤0.05). For each data set with more than two groups to compare, a one-way analysis of variance (ANOVA) was performed and the residuals tested for normality using a Shapiro-Wilk test. The normality distribution of the residuals from the second ANOVA was tested using a Shapiro-Wilk test. When the Shapiro-Wilk test was not significant (p >0.05), a Levene test was performed on the residuals to assess the homogeneity of the group variances. If differences between group variances were not found to be significant (p >0.05) then the results from the related ANOVA was retained. When significant differences among the group means were indicated by the ANOVA overall F test (p ≤0.05), a Dunnett test was used to perform the group mean comparisons between the control and each dose group. When the Shapiro-Wilk test on the residuals of the second ANOVA was significant or when heterogeneous group variances (p ≤ 0.05) are revealed by the Levene test, the groups were compared using a non-parametric Kruskal Wallis test. When the Kruskal Wallis test was significant (p ≤0.05), a Dunn test was used to perform the pairwise group comparisons between the control group and each dose group. For each data set with more than two groups to compare, data were analyzed using the Kruskal-Wallis test. When the Kruskal Wallis test was significant (p ≤0.05), the Dunn test was used to perform the pairwise group comparisons of interest. Group incidence results are presented for each considered parameter. If there are more than two groups to compare, an overall test (including all groups) was performed using the Fisher exact test. If the overall test is significant (p ≤0.05) or if there are only two groups to compare, the pairwise group comparison of the control group with each of the other dose groups were done using Fisher’s exact test.
Indices:
- Pregnancy rate (%) = (Number of pregnant rats / Number of mated rats) x 100
- Pre-implantation loss (%) = [(Number of corpora lutea - Number of implants) / Number of corpora lutea] x 100
- Post-implantation loss (%) = [(Number of implants - Number of live fetuses) / Number of implants] x 100
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Slight to severe salivation in all animals at the high dose group of 1000 mg/kg bw/day beginning on GD 9 post-dose; 2/24 mid-dose animals (300 mg/kg bw/day) were observed with slight to moderate salivation once on GD 18 and GD 20.
Dermal irritation (if dermal study):
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Statistically significant reduction in body weight change for females dosed at 1000 mg/kg bw/day following the first day of administration (GD 5 to 6) that was not considered to be of toxicological importance.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Statistically significant reduction of food consumption was noted for females on GD 6 to 9 at 1000 mg/kg bw/day. This reduction was not considered to be toxicologically significant.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
CLINICAL OBSERVATIONS
There was no mortality throughout the course of the study. Test-item related clinical signs of slight to severe salivation were observed in all animals dosed at the high dose of 1000 mg/kg bw/day beginning on GD 9 post-dose. 2/24 mid-dose animals (300 mg/kg bw/day) were observed with slight to moderate salivation once on GD 18 and GD 20, respectively and 1/24 control group animal was observed with severe salivation once on GD 15. Other clinical signs that included but were not limited to fur stained red or skin dry were considered incidental since they were observed in the control group or prior to the start of dosing.

BODY WEIGHT
There were no test item-related significant effects on body weight, body weight changes or corrected body weight changes up to 1000 mg/kg bw/day in comparison to the control group. There was a statistical significant (p ≤0.5 or p ≤0.01) reduction in body weight change noted for females dosed at 1000 mg/kg bw/day following the first day of administration (GD 5 to 6), that was related to the test item but not considered to be of toxicological importance as the weights rebounded on subsequent weighing. A slight reduction in corrected body weights changes was noted at 300 and 1000 mg/kg bw/day.

FOOD CONSUMPTION AND COMPOUND INTAKE
There were no test item-related effects on food consumption up to 1000 mg/kg bw/day in comparison to the control group. A statistical significant reduction of food consumption was noted for females on GD 6 to 9 at 1000 mg/kg bw/day. This reduction was not considered to be toxicologically significant as animal’s consumption were comparable or increased throughout the pregnancy.

POST-MORTEM EXAMINATIONS
There was no macroscopic tissue changes considered related to the administration of the test item.
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
effects observed, non-treatment-related
Description (incidence and severity):
Although all animals on study were observed as having vaginal plugs on Day 0, at the end of the study, one female of the low-dose group (100 mg/kg bw/day) was not pregnant.
Other effects:
not examined
Details on maternal toxic effects:
MATERNAL PERFORMANCE
Although all animals on study were observed as having vaginal plugs on Day 0, at the end of the study, one female of the low-dose group (100 mg/kg bw/day) was not pregnant. The pregnancy rates were 100, 95.8, 100 and 100% for the control, low-dose, mid-dose and high-dose groups, respectively.

UTERINE FINDINGS
There were no test item-related effects on gravid uterus weights, the number of corpora lutea, implantation sites, live fetuses, sex ratio, dead fetuses, resorptions or pre- and post-implantation losses.
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: no adverse effects observed
Dose descriptor:
NOEL
Remarks:
systemic
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
clinical signs
body weight and weight gain
food consumption and compound intake
Fetal body weight changes:
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): not examined
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Other effects:
not examined
Details on embryotoxic / teratogenic effects:
FETAL WEIGHTS
There were no significant test item-related effects on male, female and total (combined) fetal weights up to 1000 mg/kg bw/day.

EXTERNAL AND INTERNAL MALFORMATIONS
There were no major malformations or minor external, internal or skeletal anomalies that were considered related to treatment with the test substance at any dose level up to 1000 mg/kg bw/day.
Dose descriptor:
NOAEL
Remarks:
developmental
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
Developmental effects observed:
no
Conclusions:
The administration of the test substance by oral gavage daily from Gestation Days 5 to 20 inclusive to gravid female Sprague-Dawley rats at dose levels of 0, 100, 300, and 1000 mg/kg bw/day was well tolerated. Clinical signs such as salivation and reduction of food consumption and body weights at the high-dose group (1000 mg/kg bw/day) were recorded. However, there was no evidence of embryolethality, fetotoxicity or teratogenicity at doses up to 1000 mg/kg/day. Thus, the NOAEL for maternal and developmental toxicity was considered to be greater than 1000 mg/kg bw/day.
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:
The study is GLP according to OECD TG 414.
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

A Prenatal Developmental Toxicity Study was performed with the test substance via the oral route according to OECD guideline 414 and in compliance with GLP (2016). Groups of 24 female Sprague-Dawley rats received daily oral gavage doses of the test substance in vehicle (corn oil) at dose levels of 100, 300 and 1000 mg/kg bw/day or vehicle alone during gestational days 5 to 20. On day 21 of gestation the animals were euthanized and examined for maternal and fetal parameters. There were no mortalities or test item-related effects on clinical observations (apart from salivation and transient reduction of body weight at 1000 mg/kg/day), food consumption, body weight, body weight change, corrected body weights, gravid uterus weights or macroscopic changes at necropsy. No effects on fetal body weight, and the number of corpora lutea, implantation sites, live fetuses, sex ratio, resorptions, or pre- and post-implantation losses were observed. Furthermore, no incidences of major malformations or minor external or internal anomalies, skeletal anomalies or skeletal variants that were related to the treatment with the test substance were recorded at any dose level up to and including the highest dose level of 1000 mg/kg bw/day. Taken together, there was no evidence of embryo lethality, fetotoxicity or teratogenicity at dose levels up to 1000 mg/kg bw/day. Thus, the NOAEL for maternal/embryo-/fetotoxicity/teratogenicity in rats was found to be ≥1000 mg/kg bw/day for the test substance.

Justification for classification or non-classification

- Toxicity to reproduction:

Based on the above stated assessment of the reproduction and developmental toxicity the potential of the substance is deemed not to be toxic to the reproduction and intrauterine development and accordingly do not need to be classified according to CLP Regulation (EC) No. 1272/2008 of the European Parliament and of the Council as implementation of UN-GHS in the EU.