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Administrative data

Description of key information

LD50 (female, rat, oral): greater than 2000 mg/kg body weight.
LD50 (female/male, rat, dermal): greater than 2000 mg/kg body weight

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14-JUN-2006 to 07-JUL-2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with GLP
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Test System: Rat, HanRcc:WIST (SPF)
Number of animals per group: 3 females
Total number of animals: 6 females
Age at treatment: 12 weeks (females)
Identification: By unique cage number and corresponding color-coded spots on the tail. The animals were marked at acclimatization start.
Randomization: Selected by hand at time of delivery. No computer generated randomization program.
Acclimatization: Under laboratory conditions after health examination. Only healthy animals were used for the study.

Conditions
Standard Laboratory Conditions.
Air-conditioned with ranges for room temperature of 22 ± 3 °C and relative humidity 30-70 % and approximately 10-15 air changes per hour. 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Accommodation: during acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding.
Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet (batch 001/06) ad libitum.
Water: Community tap water, ad libitum.
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
The animals received a single dose of the test item by oral gavage administration at 2000 mg/kg body weight after being fasted for approximately 18 to 19 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing.
The application volume was 10 mL/kg body weight.
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
6 females
Control animals:
not specified
Details on study design:
Two groups, each of three female HanRcc:WIST (SPF) rats, were treated with test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) at a concentration of 0.2 g/mL and administered at a volume dosage of 10 mL/kg.
The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically. All animals survived until the end of the study period.
Statistics:
No statistical analysis was used.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Mild or moderate clinical signs of toxicity (ruffled fur, hunched posture, sedation) were noted in the animals on the day of dosing and a slightly ruffled fur persisted in two animals still on day 2. From day 3 on for the remaining observation period all animals were absent of clinical findings.
Clinical signs:
A ruffled fur was observed in all animals on the day of dosing starting 1, 2 or 3 hours after dosing and persisting up to the 5-hour reading. This symptom was graded moderate in two animals 2-5 or 3-5 hours after dosing and otherwise considered slight. A slightly ruffled fur was still present in two animals on day 2 of the study.

A hunched posture was observed in three animals on the day of dosing 0.5-5 or 2-5 hours after administration or at the 0.5-hour reading and again 2-5 hours after dosing, respectively.

In addition, three animals expressed sedation 3-5 hours after dosing. The sedation was graded moderate in one animal at the 3-hour observation and otherwise considered slight. From day 3 on up to the end of the 2-week observation period all animals remained absent of clinical findings.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necropsy
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
LD50 (female rat, oral): greater than 2000 mg/kg body weight.
Executive summary:

Two groups, each of three female HanRcc:WIST (SPF) rats, were treated with test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) at a concentration of 0.2 g/mL and administered at a volume dosage of 10 mL/kg.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period.

Mild or moderate clinical signs of toxicity (ruffled fur, hunched posture, sedation) were noted in the animals on the day of dosing and a slightly ruffled fur persisted in two animals still on day 2. From day 3 on for the remaining observation period all animals were absent of clinical findings.

The body weight of the animals was within the range commonly recorded for this strain and age.

No macroscopic findings were recorded at necropsy.

Therefore, the LD50 of test item can be estimated to be greater than 2000 mg/kg bw, which indicates that test article shall not be classified according to CLP (Regulation EC No. 1272/2008).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13-JUN-2006 to 04-JUL-2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with GLP
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Test System: Rat, HanRcc:WIST (SPF)
Number of animals per group: 5 males and 5 females
Total number of animals: 5 males and 5 females
Age at treatment: 8 weeks (male); 12 weeks (females)
Identification: By unique cage number and corresponding color-coded spots on the tail. The animals were marked at acclimatization start.
Randomization: Selected by hand at time of delivery. No computer generated randomization program.
Acclimatization: Under laboratory conditions after health examination. Only healthy animals were used for the study.

Conditions
Standard Laboratory Conditions.
Air-conditioned with ranges for room temperature of 22 ±3 °C and relative humidity 30-70 % and approximately 10-15 air changes per hour. 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Accommodation: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding during treatment and observation.
Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet (batch 001/06) ad libitum.
Water: Community tap water, ad libitum.
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
The test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
Application volume/kg body weight: 6 mL
Duration of exposure:
24 hours
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
5
Control animals:
not specified
Details on study design:
One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10 % of the total body surface.
Only those animals without injury or irritation on the skin were used in the test.
On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.
Application volume/kg body weight: 6 mL
Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels. Thereafter, the reaction sites were assessed.
Statistics:
No statistical analysis was used.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred during the study.
Clinical signs:
No systemic signs of toxicity were observed during the study period.
A purple staining of the treated skin was noted in all animals on days 2 and 3 of the study. The staining persisted still on day 4 in one female.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age with the exception of one female, which lost 1.4% of body weight during the first week after treatment. This animal recovered towards the end of the study.
Gross pathology:
No macroscopic findings were observed at necropsy.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The median lethal dose of test item after single dermal administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat, dermal): greater than 2000 mg/kg body weight
Executive summary:

Five male and five female HanRcc:WIST (SPF) rats were treated with test item at 2000 mg/kg by dermal application. The test item was diluted in vehicle (purified water) at a concentration of 0.33 g/mL and administered at a volume dosage of 6 ml/kg. The application period was 24 hours.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Local signs were noted once daily from test day 2 to 15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

No deaths occurred during the study.

No clinical signs were observed during the course of the study.

The treated skin area was stained purple in all animals on days 2 and 3 after treatment. The staining persisted still on day 4 in one female. Otherwise, no dermal symptoms were noted. The body weight of the animals was within the range commonly recorded for this strain and age with the exception of one female, which lost 1.4% of body weight during the first week after treatment. This animal recovered towards the end of the study.

No macroscopic findings were observed at necropsy.

Therefore, the LD50 of the test article was estimated to be greater than 2000 mg/kg.bw, which indicates that test article shall not be classified in accordance with CLP (Regulaton EC No. 1272/2008).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Oral toxicity test and dermal toxicity test of the test substance were performed under GLP test condition according to the OECD guideline. The inhalation toxicity test is waived according to column 2 of REACH Annex VIII.

Two groups, each of three female HanRcc:WIST (SPF) rats, were treated with test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was diluted in vehicle (purified water) at a concentration of 0.2 g/mL and administered at a volume dosage of 10 mL/kg.All animals survived until the end of the study period.Mild or moderate clinical signs of toxicity (ruffled fur, hunched posture, sedation) were noted in the animals on the day of dosing and a slightly ruffled fur persisted in two animals still on day 2. From day 3 on for the remaining observation period all animals were absent of clinical findings.The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy. Therefore, the LD50 of test item can be estimated to be greater than 2000 mg/kg bw.

Five male and five female HanRcc:WIST (SPF) rats were treated with test item at 2000 mg/kg by dermal application. The test item was diluted in vehicle (purified water) at a concentration of 0.33 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours. No deaths occurred during the study. No clinical signs were observed during the course of the study. The treated skin area was stained purple in all animals on days 2 and 3 after treatment. The staining persisted still on day 4 in one female. Otherwise, no dermal symptoms were noted. The body weight of the animals was within the range commonly recorded for this strain and age with the exception of one female, which lost 1.4 % of body weight during the first week after treatment. This animal recovered towards the end of the study. No macroscopic findings were observed at necropsy. Therefore, the LD50 of test article was estimated to be greater than 2000 mg/kg.bw.


Justification for selection of acute toxicity – oral endpoint
guidance test with GLP compliance

Justification for selection of acute toxicity – dermal endpoint
guidance test with GLP compliance

Justification for classification or non-classification

Based on the acute oral and dermal toxicity test results with no mortality observed at 2000 mg/kg bw and no specific target organ effects seen, the test substance is not classified according to the CLP regulation (Regulation EC No. 1272/2008) and also not according to DSD (Directive 67/548/EEC) for acute toxicity or specific target organ toxicity, single exposure.