Registration Dossier

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Toxicity to Reproduction:

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-reprotoxic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Weight of evidence approach based on various test chemicals
Justification for type of information:
Weight of evidence approach based on various test chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: Weight of evidence approach based on various test chemicals
Principles of method if other than guideline:
Weight of evidence approach based on various test chemicals
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 2. Wistar; 3. long-Evans
Sex:
male/female
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 2. water; 3. diet
Details on exposure:
2.PREPARATION OF DOSING SOLUTIONS: The dosing solutions were prepared at concentrations 0,250, 500 and 1000 mg/kg such that a volume of 10 ml/kg was administered daily.
DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: 0, 250, 500 and 1000 mg/kg/day
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available
3. no data available
Details on mating procedure:
2. Details on mating procedure
- M/F ratio per cage: 1 : 1 ratio
- Length of cohabitation: Overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pr
egnancy Sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: This procedure was repeated over several consecutive days until at least 30 females had been allocated to each treatment group.
3. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selectedanimals (10 males,20 females) from the F1b litters were used for breeding.Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
2. 19 days
3. 80-day ( 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations.)
Frequency of treatment:
2. daily
3. daily
Details on study schedule:
2. Dose selection rationale: No data available
- Rationale for animal assignment (if not random): The day of sperm detection was designated day 0 of pregnancy and the mated females were randomly allocated to one of four treatment groups. This procedure was repeated over several consecutive days until at least 30 females had been allocated to each treatment group.
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
3. - F1 parental animals not mated until [...] weeks after selected from the F1 litters.The F0 generation rats were mated twice,
- Selection of parents from F1 generation when pups were [...] days of age.:the F1a
litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b
litters were used for breeding.Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to theF1a and F1b generations.
- Age at mating of the mated animals in the study: [...] weeks:80days
- Section schedule rationale (if not random): No data available
Remarks:
Study 2: 0, 250, 500 and 1000 mg/kg/day
Remarks:
study 3. 0, 10, 100, 300, or 1,000 mg/kg b.w./day
No. of animals per sex per dose:
2. Total: 120
0 mg/kg/day: 30 female
250 mg/kg/day: 30 female
500 mg/kg/day: 30 female
1000 mg/kg/day: 30 female
3. Total:300
F0 generation
0 mg/kg bw/day:10male and 20 females
10mg/kg bw/day:10male and 20 females
100mg/kg bw/day:10male and 20 females
300 mg/kg bw/day:10male and 20 females
1000mg/kg bw/day: 10male and 20 females
F1 generation
0 mg/kg bw/day:10male and 20 females
10mg/kg bw/day:10male and 20 females
100mg/kg bw/day:10male and 20 females
300 mg/kg bw/day:10male and 20 females
1000mg/kg bw/day: 10male and 20 females
Control animals:
yes, concurrent vehicle
Details on study design:
no data available
Positive control:
no data available
Parental animals: Observations and examinations:
2.BODY WEIGHT: Yes / No / No data: yes
- Time schedule for examinations:Each animal was weighed on alternate days, commencing on day 0 of pregnancy.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes / No / No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g foo
d/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data

POST-MORTEM EXAMINATIONS: Yes / No / No data: yes
- Sacrifice on gestation day # : On day 20 of pregnancy, 30 females from each group were killed by c
ervical dislocation and a postmortem examination was carried out
- Organs examined: a postmortem examination was carried out, during which the gross appearance of the maternal tissues was noted and the foetuses were removed. The numbers of corpora lutea and implantation sites and the numbers and positions of the sites with dead, live or resorbed foetuses were recorded.
OTHER: At 7 wk of age vaginal smears were prepared and examined for each female. Those females in pro-oestrus (indicated by the presence of mainly cornified and epithelial cells) were caged
individually overnight with a mature male.
3.CAGE SIDE OBSERVATIONS: Yes
- Time schedule: no data available
- Cage side observations checked in table [No.?] were included.
DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule: no data available
BODY WEIGHT: Yes
- Time schedule for examinations:
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations:
Oestrous cyclicity (parental animals):
2. corpora lutea per ovary was examined.
3. corpora lutea per ovary was examined.
Sperm parameters (parental animals):
2. no data available
Litter observations:
2.Weight of fetuses, Dead, live or resorbed fetuses were recorded.
Postmortem examinations (parental animals):
2. Gross appearances were observed.
3. SACRIFICE: After third mating, half of the pregnant dams were sacrificed on day 19 of gestation.
GROSS NECROPSY : Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning.
HISTOPATHOLOGY / ORGAN WEIGHTS : stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals,kidneys, urinary bladder, thyroid, ovaries, and uterus or testes was examined histologically.
Postmortem examinations (offspring):
2.
- External examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Soft tissue examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Skeletal examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Head examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: no data available
3. GROSS NECROPSY:yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]: Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning.

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals,kidneys, urinary bladder, thyroid, ovaries, and uterus or testes was examined histologically.
Reproductive indices:
2. Number of corpora lutea, implantation sites, Pre-implantation losses, Early resorptions, Late resorptions and Post-implantation losses was recorded.
3. pregnancy and fertility rates, gestation length
Offspring viability indices:
2. Viability of fetuses were observed.
3. offspring survival,weights and sex, litter survival,resorption rates
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
3. no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. No statistically significant differences were observed in treated female rats as compared to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control.
3. No effects attributable to treatment were observed with respect to mating performance,pregnancy and fertility rates, gestation length.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control.
3. No effects attributable to treatment were observed with respect to mating performance,pregnancy and fertility rates, gestation length.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
gross pathology
reproductive function (oestrous cycle)
reproductive performance
Critical effects observed:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
3. No effects attributable to treatment were observed with respect to adult mortality, offspring survival.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to body weight.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
3. No changes attributable to treatment were observed with respect to food consumption.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Gross pathological findings:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain
organ weights and organ / body weight ratios
reproductive function (oestrous cycle)
reproductive performance
Critical effects observed:
no
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
2. No effect were observed on Live foetuses treated female rats as compared to control
3. No effects attributable to treatment were observed with respect to adult mortality, offspring survival.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. No significant effect was observed on fetuses of treated female rats as compared to control.
3. No effects attributable to treatment were observed with respect to body weight
Food consumption and compound intake (if feeding study):
not specified
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
2. Statistically significant increase in fetuses with mucus in tracea were observed in 250, 500 and 1000mg/kg/day treated female rats as compared to control.This findings were observed at all dose levels and the incidence was not dose related.Ossification of proximal phalanges and fourth metacarpals of Skeletal tissue were observed in 500 and 1000 mg/kg/day treated fetuses as compared to control. Despite this association with treatment, the nature of thefinding does not indicate an adverse effect.
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain
gross pathology
Critical effects observed:
no
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to adult mortality,offspring survival
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Gross pathological findings:
not specified
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Histopathological findings:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance,pregnancy and fertility rates, gestation length, offspring
survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F2a
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
histopathology: non-neoplastic
Dose descriptor:
NOAEL
Generation:
F2b
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
gross pathology
histopathology: non-neoplastic
Critical effects observed:
not specified
System:
other: not specified
Reproductive effects observed:
no
Treatment related:
no
Conclusions:
Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-reprotoxic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.
Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical. The studies are as mentioned below:

Teratogenicity and embryotoxicity study of the test chemical was performed on rats. Daily oral doses of 0 (control), 250, 500 or 1000 mg/kg body weight were given to groups of 30 pregnant Wistar rats on days 0-19 of pregnancy.The dosing solutions were prepared at concentrations such that a volume of 10ml/kg was administered daily. The controls received water. Each animal was weighed on alternate days, commencing on day 0 of pregnancy. On day 20 of pregnancy, 30 females from each group were killed by cervical dislocation and a postmortem examination was carried out, during which the gross appearance of the maternal tissues was noted and the foetuses were removed. The numbers of corpora lutea and implantation sites and the numbers and positions of the sites with dead, live or resorbed fetuses were recorded. The live foetuses were weighed and examined for gross abnormalities.This treatment did not adversely influence maternal body weight, the numbers of implanations, pre- or post implantation losses or of live foetuses, the sex ratio or the weight of the litters or foetuses. No definite abnormalities were seen and the only finding in the examination of stained skeletons was a slightly more advanced ossification of the forelimbs of the offspring from females given 500 or 1000 mg /kg/day. More foetuses with mucus in the trachea were found in the treated groups than in the controls but this was not considered to be a teratogenic effect. Thus no embryotoxic or teratogenic effects were detected with doses of up to 1000 mg/kg/day throughout pregnancy.

This is supported by the results of the 3-generation reproduction study carried out on the test chemical in male and female Long-Evans rats. The test material was mixed with feed and administered at dose levels of 0, 10, 100, 300, or 1,000 mg/kg b.w./day. The first generation parents (10 males, 20 females) were given the appropriate dose of test material in the diet 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used for breeding. Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations. Following the third mating, half of the pregnant dams were sacrificed on day 19 of gestation, the uterine contents were examined for total embryos/resorption sites, and the corpora lutea per ovary were recorded. The other half were allowed to deliver normally (F2c) and were sacrificed at weaning. The F2b animals were mated once and allowed to raise their offspring to weaning when both parents and offspring were culled.Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning. Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment. The No observed adverse effect level (NOAEL) for reproductive toxicity was considered to be 1000 mg/kg bw /day,as  there were no adverse effects on food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorptions rates, or necropsy findings at dose concentration 1000 mg/kg bw /day.

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-reprotoxic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Experimental exposure time per week (hours/week):
168
Species:
rat
Quality of whole database:
Klimisch Rating 2
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Data available from different studies were reviewed to determine the reproductive toxicity of test chemical. The studies are as mentioned below:

Teratogenicity and embryotoxicity study of the test chemical was performed on rats. Daily oral doses of 0 (control), 250, 500 or 1000 mg/kg body weight were given to groups of 30 pregnant Wistar rats on days 0-19 of pregnancy.The dosing solutions were prepared at concentrations such that a volume of 10ml/kg was administered daily. The controls received water. Each animal was weighed on alternate days, commencing on day 0 of pregnancy. On day 20 of pregnancy, 30 females from each group were killed by cervical dislocation and a postmortem examination was carried out, during which the gross appearance of the maternal tissues was noted and the foetuses were removed. The numbers of corpora lutea and implantation sites and the numbers and positions of the sites with dead, live or resorbed fetuses were recorded. The live foetuses were weighed and examined for gross abnormalities.This treatment did not adversely influence maternal body weight, the numbers of implanations, pre- or post implantation losses or of live foetuses, the sex ratio or the weight of the litters or foetuses. No definite abnormalities were seen and the only finding in the examination of stained skeletons was a slightly more advanced ossification of the forelimbs of the offspring from females given 500 or 1000 mg /kg/day. More foetuses with mucus in the trachea were found in the treated groups than in the controls but this was not considered to be a teratogenic effect. Thus no embryotoxic or teratogenic effects were detected with doses of up to 1000 mg/kg/day throughout pregnancy.

This is supported by the results of the 3-generation reproduction study carried out on the test chemical in male and female Long-Evans rats. The test material was mixed with feed and administered at dose levels of 0, 10, 100, 300, or 1,000 mg/kg b.w./day. The first generation parents (10 males, 20 females) were given the appropriate dose of test material in the diet 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used for breeding. Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations. Following the third mating, half of the pregnant dams were sacrificed on day 19 of gestation, the uterine contents were examined for total embryos/resorption sites, and the corpora lutea per ovary were recorded. The other half were allowed to deliver normally (F2c) and were sacrificed at weaning. The F2b animals were mated once and allowed to raise their offspring to weaning when both parents and offspring were culled.Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning. Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment. The No observed adverse effect level (NOAEL) for reproductive toxicity was considered to be 1000 mg/kg bw /day,as  there were no adverse effects on food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorptions rates, or necropsy findings at dose concentration of 1000 mg/kg bw /day.

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-reprotoxic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.

Effects on developmental toxicity

Description of key information

Developmental Toxicity:

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-teratogenic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Weight of evidence approach based on various test chemicals
Justification for type of information:
Weight of evidence approach based on various test chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: Weight of evidence approach based on various test chemicals
Principles of method if other than guideline:
Weight of evidence approach based on various test chemicals
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 2. Wistar; 3. Long-Evans
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 2. water; 3.
Details on exposure:
2. PREPARATION OF DOSING SOLUTIONS: The dosing solutions were prepared at concentrations 0, 250, 500 and 1000 mg/kg such that a volume of 10 ml/kg was administered daily.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: 0, 250, 500 and 1000 mg/kg/day
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available
3.
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
2. - M/F ratio per cage: 1 : 1 ratio
- Length of cohabitation: Overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): Individually
- Any other deviations from standard protocol: This procedure was repeated over several consecutive days until at least 30 females had been allocated to each treatment group.
3. - Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]: cohoused
- If cohoused: yes
- M/F ratio per cage: 1:2
- Other: The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males,20 females) from the F1b litters were used for breeding.Following an 80-
day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations
- F1 parental animals not mated until [...] weeks after selected from the F1 litters.The F0 generation rats were mated twice,
- Selection of parents from F1 generation when pups were [...] days of age.:the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used
for breeding.Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b
generations.
- Age at mating of the mated animals in the study: [...] weeks:80 days
Duration of treatment / exposure:
2. 19 days
3. 80-day (2 weeks before the first mating, and dosing continued throughout the gestation, lactation,andpost-weaning phases for three successive generations)
Frequency of treatment:
2. daily
3. daily
Duration of test:
2. 19 days
3. 80-day (2 weeks before the first mating, and dosing continued throughout the gestation, lactation,and post-weaning phases for three successive generations)
Remarks:
Study 2: 0, 250, 500 and 1000 mg/kg bw/day
Remarks:
Study 3: 0, 10, 100, 300, or 1,000 mg/kg b.w./day
No. of animals per sex per dose:
2. Total: 120
0 mg/kg/day: 30 female
250 mg/kg/day: 30 female
500 mg/kg/day: 30 female
1000 mg/kg/day: 30 female
3. Total:300
F0 generation
0 mg/kg bw/day:10male and 20 females
10mg/kg bw/day:10male and 20 females
100mg/kg bw/day:10male and 20 females
300 mg/kg bw/day:10male and 20 females
1000mg/kg bw/day: 10male and 20 females
F1 generation
0 mg/kg bw/day:10male and 20 females
10mg/kg bw/day:10male and 20 females
100mg/kg bw/day:10male and 20 females
300 mg/kg bw/day:10male and 20 females
1000mg/kg bw/day: 10male and 20 females
Control animals:
yes, concurrent vehicle
Maternal examinations:
2. BODY WEIGHT: Yes / No / No data: yes
- Time schedule for examinations:Each animal was weighed on alternate days, commencing on day 0 of pregnancy.

POST-MORTEM EXAMINATIONS: Yes / No / No data: yes
- Sacrifice on gestation day # : On day 20 of pregnancy, 30 females from each group were killed by cervical dislocation and a postmortem examination was carried out
- Organs examined: a postmortem examination was carried out, during which the gross appearance of the maternal tissues was noted and the foetuses were removed. The numbers of corpora lutea and implantation sites and the numbers and positions of the sites with dead, live or resorbed foetuses were recorded.
OTHER: At 7 wk of age vaginal smears were prepared and examined for each female. Those females in pro-oestrus (indicated by the presence of mainly cornified and epithelial cells) were caged
individually overnight with a mature male.

3. CAGE SIDE OBSERVATIONS: Yes / No / No data: yes
- Time schedule: throughout the study
- Cage side observations checked in table [No.?] were included. : adult mortality and offspring mortality was observed for all generations.

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data: no data available
- Time schedule: no data available

BODY WEIGHT: Yes / No / No data: yes
- Time schedule for examinations: throughout the study.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data : yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data: no data available
- Time schedule for examinations: no data available

POST-MORTEM EXAMINATIONS: Yes / No / No data: yes
- Sacrifice on gestation day # : Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning.
- Organs examined: Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes
Ovaries and uterine content:
2. The ovaries and uterine content was examined after termination: Yes / No / No data: yes
Examinations included:
- Gravid uterus weight: Yes / No / No data: yes
- Number of corpora lutea: Yes / No / No data: yes
- Number of implantations: Yes / No / No data: yes
- Number of early resorptions: Yes / No / No data: yes
- Number of late resorptions: Yes / No / No data: yes
3. The ovaries and uterine content was examined after termination: Yes / No / No data: yes
Examinations included:
- Number of corpora lutea: Yes / No / No data: yes
- Number of implantations: Yes / No / No data: yes
- Number of early resorptions: Yes / No / No data: yes
- Number of late resorptions: Yes / No / No data: yes
Fetal examinations:
2. - External examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data:yes
- Soft tissue examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Skeletal examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Head examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
3.- External examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data:yes
- Soft tissue examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Skeletal examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
- Head examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data: yes
Statistics:
no data available
Indices:
2. Viability of fetuses were observed.
3. pregnancy and fertility rates, gestation length
Historical control data:
no data available
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
3. No effects attributable to treatment were observed with respect to adult mortality
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. There were no statistically significant differences in body weight between treated and control females
3. No effects attributable to treatment were observed with respect to body weight
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
3. no effects attributable to treatment were observed with respect to food consumption
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
2. At autopsy, all the maternal animals given the test chemical were found to have colouring in the gastro-intestinal tract and the placental tissue also appeared to the slightly green. Apart from this,
no abnormalities were found in the tissues of the maternal animals.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
2. There were no differences between the control and treated groups in the results of the examination of the reproductive system and the foetuses
3. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control.
Number of abortions:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control
3.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control
3. No effects attributable to treatment were observed with respect to resorption rates.
Early or late resorptions:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control
3. No effects attributable to treatment were observed with respect to resorption rates.
Dead fetuses:
no effects observed
Description (incidence and severity):
2. No effect were observed on number of corpora lutea, implantation sites, Pre-implantation losses, early resorptions, late resorptions and post-implantation losses of treated female rats as compared to control
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to gestation length
Changes in number of pregnant:
not specified
Other effects:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
early or late resorptions
effects on pregnancy duration
histopathology: non-neoplastic
maternal abnormalities
necropsy findings
number of abortions
pre and post implantation loss
total litter losses by resorption
Abnormalities:
no effects observed
Fetal body weight changes:
not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
3. No effects attributable to treatment were observed with respect to offspring survival
External malformations:
no effects observed
Description (incidence and severity):
2. a lack of teratogenic effect was indicated by the absence of any gross malformations of severe defects in the examination of the skeletal or soft parts.
3. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment
Skeletal malformations:
no effects observed
Description (incidence and severity):
2. a lack of teratogenic effect was indicated by the absence of any gross malformations of severe defects in the examination of the skeletal or soft parts.
3. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment
Visceral malformations:
no effects observed
Description (incidence and severity):
2. a lack of teratogenic effect was indicated by the absence of any gross malformations of severe defects in the examination of the skeletal or soft parts.
3.There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment
Other effects:
no effects observed
Description (incidence and severity):
2. Ossification of proximal phalanges and fourth metacarpals of Skeletal tissue were observed in 500 and 1000 mg/kg/day treated fetoses as compared to contorl. Despite this association with treatment, the nature of the finding does not indicate an adverse effect.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reduction in number of live offspring
external malformations
skeletal malformations
visceral malformations
Abnormalities:
no effects observed
Developmental effects observed:
not specified
Conclusions:
Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-teratogenic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.
Executive summary:

Data available from different studies were reviewed to determine the teratogenic toxicity potential of test chemical. The studies are as mentioned below:

Teratogenicity and embryotoxicity study of the test chemical was performed on rats. Daily oral doses of 0 (control), 250, 500 or 1000 mg/kg body weight were given to groups of 30 pregnant Wistar rats on days 0-19 of pregnancy.The dosing solutions were prepared at concentrations such that a volume of 10ml/kg was administered daily. The controls received water. Each animal was weighed on alternate days, commencing on day 0 of pregnancy. On day 20 of pregnancy, 30 females from each group were killed by cervical dislocation and a postmortem examination was carried out, during which the gross appearance of the maternal tissues was noted and the foetuses were removed. The numbers of corpora lutea and implantation sites and the numbers and positions of the sites with dead, live or resorbed foetuses were recorded. The live foetuses were weighed and examined for gross abnormalities.This treatment did not adversely influence maternal body weight, the numbers of implanations, pre- or post-implantation losses or of live foetuses, the sex ratio or the weight of the litters or foetuses. No definite abnormalities were seen and the only finding in the examination of stained skeletons was a slightly more advanced ossification of the forelimbs of the offspring from females given 500 or 1000 mg /kg/day. More foetuses with mucus in the trachea were found in the treated groups than in the controls but this was not considered to be a teratogenic effect. Thus no embryotoxic or teratogenic effects were detected with doses of up to I000 mg/kg/day throughout pregnancy

This is supported by the results of the 3-generation reproduction study carried out on the test chemical in male and female Long-Evans rats. The test material mixed with feed and administered at dose levels of 0, 10, 100, 300, or 1,000 mg/kg b.w./day. The first generation parents (10 males, 20 females) were given the appropriate dose of test material in the diet 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used for breeding. Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations. Following the third mating, half of the pregnant dams were sacrificed on day 19 of gestation, the uterine contents were examined for total embryos/resorption sites, and the corpora lutea per ovary were recorded. The other half were allowed to deliver normally (F2c) and were sacrificed at weaning. The F2b animals were mated once and allowed to raise their offspring to weaning when both parents and offspring were culled.Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning. Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment. The No observed adverse effect level (NOAEL) for reproductive and developmental toxicity was considered to be 1000 mg/kg bw /day,as  there were no adverse effects on food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorptions rates, or necropsy findings were observed in rat at dose concentration 1000 mg/kg bw /day.

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-teratogenic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/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:
subchronic
Experimental exposure time per week (hours/week):
168
Species:
rat
Quality of whole database:
Klimisch rating 2
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

Data available from different studies were reviewed to determine the teratogenic toxicity potential of test chemical. The studies are as mentioned below:

Teratogenicity and embryotoxicity study of the test chemical was performed on rats. Daily oral doses of 0 (control), 250, 500 or 1000 mg/kg body weight were given to groups of 30 pregnant Wistar rats on days 0-19 of pregnancy.The dosing solutions were prepared at concentrations such that a volume of 10ml/kg was administered daily. The controls received water. Each animal was weighed on alternate days, commencing on day 0 of pregnancy. On day 20 of pregnancy, 30 females from each group were killed by cervical dislocation and a postmortem examination was carried out, during which the gross appearance of the maternal tissues was noted and the foetuses were removed. The numbers of corpora lutea and implantation sites and the numbers and positions of the sites with dead, live or resorbed foetuses were recorded. The live foetuses were weighed and examined for gross abnormalities.This treatment did not adversely influence maternal body weight, the numbers of implanations, pre- or post-implantation losses or of live foetuses, the sex ratio or the weight of the litters or foetuses. No definite abnormalities were seen and the only finding in the examination of stained skeletons was a slightly more advanced ossification of the forelimbs of the offspring from females given 500 or 1000 mg /kg/day. More foetuses with mucus in the trachea were found in the treated groups than in the controls but this was not considered to be a teratogenic effect. Thus no embryotoxic or teratogenic effects were detected with doses of up to 1000 mg/kg/day throughout pregnancy

This is supported by the results of the 3-generation reproduction study carried out on the test chemical in male and female Long-Evans rats. The test material wasmixed with feed and administered at dose levels of 0, 10, 100, 300, or 1,000 mg/kg b.w./day. The first generation parents (10 males, 20 females) were given the appropriate dose of test material in the diet 2 weeks before the first mating, and dosing continued throughout the gestation, lactation, and post-weaning phases for three successive generations. The F0 generation rats were mated twice, the F1a litters being necropsied at weaning, and selected animals (10 males, 20 females) from the F1b litters were used for breeding. Following an 80-day growth period, animals from the F1b generation were mated 3 times and the offspring of the F2a and F2b generations were treated identically to the F1a and F1b generations. Following the third mating, half of the pregnant dams were sacrificed on day 19 of gestation, the uterine contents were examined for total embryos/resorption sites, and the corpora lutea per ovary were recorded. The other half were allowed to deliver normally (F2c) and were sacrificed at weaning. The F2b animals were mated once and allowed to raise their offspring to weaning when both parents and offspring were culled.Gross necropsies were performed on all parent animals and on F1a, F2a, F2c, and F3a offspring at weaning. Selected tissues from 5 animals of each sex/dose from the F1b parents and the F3a generation at weaning were fixed at necropsy, and the following tissues examined histologically from the control and high-dose group: stomach, ileum, jejunum, colon, liver, spleen, heart, lungs, adrenals, kidneys, urinary bladder, thyroid, ovaries, and uterus or testes. No effects attributable to treatment were observed with respect to food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorption rates, or necropsy findings. There were no macroscopic or microscopic tissue abnormalities of either F1b- or F3a-generation animals considered to be attributable to treatment.

The No observed adverse effect level (NOAEL) for reproductive and developmental toxicity was considered to be 1000 mg/kg bw /day,as  there were no adverse effects on food consumption, body weight, adult mortality, mating performance, pregnancy and fertility rates, gestation length, offspring survival, weights and sex, litter survival, resorptions rates, or necropsy findings were observed in rat at dose concentration 1000 mg/kg bw /day.

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-teratogenic when exposed to the test chemical orally. The NOAEL value can be considered to be 1000 mg/kg/day.

Justification for classification or non-classification

Based on the available results and applying the weight of evidence approach, the test chemical can be considered to non-reprotoxic and non-teratogenic when exposed to the test chemical orally. Hence, the test chemical can be classified under the category "Not Classified" as per CLP Regulation.

Additional information