Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available

Effects on developmental toxicity

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Refer to "any other information on materials and methods" for additional information.
Qualifier:
according to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Refer to "any other information on materials and methods" for additional information.
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Refer to "any other information on materials and methods" for additional information.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl:CD®(SD)IGS BR
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: 63 days old
- Weight at study initiation:200-225 g
- Fasting period before study: No
- Housing: Individually in suspended, wire-mesh, stainless steel cages. Nesting material was not provided because the dams were euthanized prior to parturition.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Rats were received at 1, 2, or 3 days of gestation and were released for the study upon approval of the Laboratory Animal Veterinarian designee

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26°C
- Humidity (%): 30-70% (targeted at 40-60%)
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Formulations of the test substance in the vehicle were prepared daily.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
At the beginning of the study, dosing formulations containing the test substance at the concentrations of 2.50, 15.0, and 100.0 mg active ingredient/mL were collected. These samples were analysed to determine homogeneity/concentration verification and 5-hour room temperature stability. Near the end of the in-life phase, dosing formulations at the same levels were collected and analysed for concentration verification. A 0 mg/mL (control) sample was collected and submitted with each sampling. Concentrations of the active ingredient (a.i.) in the test substance in the formulations were determined by high-performance liquid chromatography (HPLC) with mass spectroscopy (MS) detection. The samples were analysed on the day the samples were collected. On days samples were taken, the formulations remaining after dosing were stored in the refrigerator for possible additional analysis.

Data from the analysis of the samples at the start of the study indicate that the test substance was mixed uniformly, was at the targeted levels, and stable in the vehicle under the conditions of study. The data for the concentration verification indicated that the test substance was mixed uniformly in the vehicle and at the targeted concentration during the study. Test substance was not found in the 0 mg/mL samples.
Details on mating procedure:
Nulliparous time-mated females were received for this study from Charles River Laboratories, Inc., Raleigh, North Carolina. The rats for this study were requested to be 63 days old and be at either 1, 2, or 3 days of gestation upon arrival.
Duration of treatment / exposure:
Gestation days 6-10
Frequency of treatment:
Daily
Duration of test:
Females were sacrificed on gestation day 21
Dose / conc.:
25 mg/kg bw/day
Dose / conc.:
150 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
22 females/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In a previous pilot developmental toxicity study in rats, the test substance was administered once daily by gavage to groups of time-pregnant rats (8/group) on days 6-20G at dosages of 0, 40, 200, or 1000 mg active ingredient/kg/day. There was neither mortality nor were there any test substance-related clinical signs of toxicity or gross postmortem findings in dams. Maternal toxicity was observed at 200 and 1000 mg/kg/day, and consisted of reduced body weight gain (200 and 1000 mg/kg/day) and food consumption (1000 mg/kg/day). Reproductive parameters (foetal viability, sex ratio, and foetal weight) were similar across groups. There were no test substance-related foetal external alterations at any dosage. Based on these data, the dose levels for the current study were 0, 25, 150, and 1000 mg/kg/day.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Clinical observations were recorded on the day after arrival and daily until the end of the study.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Clinical signs were recorded twice daily on days 6-20G; clinical signs observed at other times were recorded by exception.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded on the day after arrival and daily until the end of the study.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Food consumption was measured on days 4, 6, 8, 10, 12, 14, 16, 18, 20, and 21G.

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

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 21
- Organs examined: The abdominal and thoracic viscera were examined grossly immediately after euthanasia on day 21G.
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
- Other: The uterus of each apparently "non-pregnant" female was stained with ammonium sulphide to detect very early resorptions.
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: all per litter
- Head examinations: Yes: half per litter
Statistics:
Descriptive statistics (mean ± standard deviation) were calculated for all relevant endpoints listed in the statistical analysis table. For litter parameters, the proportion of affected foetuses per litter or the litter mean was used as the experimental unit for statistical evaluation. The level of significance selected was p < 0.05.
Historical control data:
The test facility maintains historical control data from 21 studies; 1998-2004.
Clinical signs:
no effects observed
Description (incidence and severity):
No test substance-related clinical observations were observed in dams at any dose level; all observations were common to this strain and age.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
A reduction in body weight gain was observed at 1000 mg/kg/day on days 6-8G (81% lower than control group mean), resulting in overall (minus products of conception) body weight gain (days 6-21G) that was 17% lower than the control group mean. Weight gain for all other periods for the 1000 mg/kg/day group and for all other treated groups were similar to control weight gains. There were no test substance-related statistically significant differences in daily maternal body weights at any dose level.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Maternal food consumption was reduced at 1000 mg/kg/day on days 6-8G (14% lower than the control group mean). This reduction in food consumption was not considered adverse since it was small, transient, and did not affect overall food consumption.
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not specified
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:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no test substance related gross postmortem findings at any dose level. Findings were limited to large and/or fused placenta(s) in 1-2 dams at 25 mg/kg/day. These findings were not considered test substance-related due to the low incidence and lack of dose response relationship.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
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:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Key result
Dose descriptor:
NOEL
Effect level:
150 mg/kg bw/day (actual dose received)
Basis for effect level:
body weight and weight gain
Key result
Abnormalities:
no effects observed
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): no effects observed
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
Description (incidence and severity):
There were no test substance-related fetal malformations observed at any dose level. One fetus in the 1000 mg/kg/day group had a vertebrae malformation (centrum displaced dorsally).
Visceral malformations:
no effects observed
Description (incidence and severity):
There were no test substance-related fetal malformations observed at any dose level. One fetus in the 25 mg/kg/day group had a heart malformation (situs inversus).
Other effects:
no effects observed
Description (incidence and severity):
There were no test substance-related variations observed at any dose level. The number and type of variations observed were similar for all groups, and were common to this strain and developmental age.
A slightly higher incidence of ribs with extra ossification sites was observed in the 150 mg/kg/day (8 litters;18 fetuses affected) and 1000 mg/kg/day (7 litters;17 fetuses affected) groups compared to the control group (3 litters;6 fetuses affected). This apparently higher incidence was not considered test substance-related since the percent affected fetuses per litter was not statistically significantly increased compared to the control group (p = 0.41 and 0.32 for the 150 and 1000 mg/kg/day groups, respectively). Furthermore, the incidence of this finding was within historical control ranges of Haskell Laboratory data (171 fetuses; 96 litters affected; 21 studies; 1998-2004). In addition, this skeletal variation is not permanent and is not considered indicative of adverse developmental toxicity.
Details on embryotoxic / teratogenic effects:
No effects observed
Key result
Dose descriptor:
NOEL
Effect level:
> 1 000 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: no effects at the highest dose level tested
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Conclusions:
Under the conditions of the study, the no-observed-effect level (NOEL) for maternal toxicity was 150 mg/kg/day, based on reduced maternal body weight gain at 1000 mg/kg/day. The NOEL for developmental toxicity was 1000 mg/kg/day, the highest dose level tested.

The study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).
Executive summary:

Groups of 22 time-mated Crl:CD®(SD)IGS BR rats were administered daily dosages of the test substance at 0, 25, 150, or 1000 mg/kg/day during gestation days 6-20 (day 6-20G). Controls were administered the vehicle only (NANOpure® water). During the in-life portion of the study, maternal body weights, food consumption, and clinical signs data were collected. On day 21G, dams were euthanized and subjected to a gross external and internal examination. Uterine contents were described; all foetuses were removed and individually weighed, sexed, and examined for external alterations. Approximately one-half of the foetuses were subjected to visceral and head evaluations; all foetuses were examined for skeletal alterations.

No maternal mortality occurred during the study. No test substance-related clinical observations were observed in dams at any level. A reduction in body weight gain was observed at 1000 mg/kg/day on days 6-8G (81% lower than control group mean), resulting in overall body weight gain (days 6-21G) that was 17% lower than the control group mean. Maternal food consumption was reduced at 1000 mg/kg/day on days 6-8G (14% lower than the control group mean). This reduction in food consumption was not considered adverse since it was small, transient, and did not affect overall food consumption. There were no test substance-related gross postmortem observations in dams at any level. The mean number of implantation sites, resorptions, and live foetuses, and the mean sex ratio and foetal weight were comparable across all groups. There were no test substance-related foetal external, visceral, or skeletal malformations or variations observed at any dose level.

Under the conditions of the study, the no-observed-effect level (NOEL) for maternal toxicity was 150 mg/kg/day, based on reduced maternal body weight gain at 1000 mg/kg/day. The NOEL for developmental toxicity was 1000 mg/kg/day, the highest dose level tested.

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
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 GLP prenatal developmental study was conducted in groups of 22 time-mated Crl:CD®(SD)IGS BR rats at daily dosages of the test substance of 0, 25, 150, or 1000 mg/kg/day during gestation days 6-20 (day 6-20G) according to OECD guideline 414. No maternal mortality or test substance-related clinical observations or gross post-mortem observations were observed in dams at any level. Test substance-related maternal toxicity was evident at 1000 mg/kg and was demonstrated as a reduction in body weight gain on days 6-8G (81% lower than control group mean), resulting in overall body weight gain (days 6-21G) that was 17% lower than the control group mean. No test substance-related effects on number of implantation sites, resorptions, live foetuses, mean sex ratio, foetal weight, foetal external, visceral, or skeletal malformations or variations were observed at any dose level. Under the conditions of the study, the no-observed-effect level (NOEL) for maternal toxicity was 150 mg/kg/day and the NOEL for developmental toxicity was 1000 mg/kg/day, the highest dose level tested.


Justification for selection of Effect on developmental toxicity: via oral route:
OECD guideline, GLP study

Justification for classification or non-classification

No test substance-related unique developmental toxicity was observed after exposure to the test substance. Therefore, the substance does not need to be classified for developmental toxicity according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

Additional information