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Administrative data

Description of key information

Acute Oral Toxicity

Under the conditions of the study, the acute oral LD50 value of the test material was found to be between 300 and 2 000 mg/kg bw in female rats.

Acute Dermal Toxicity

Under the conditions of the study the acute dermal LD50 of the test material was determined to be greater than 2 000 mg/kg body weight in female rats.

Acute Inhalation Toxicity

In accordance with Column 2 of REACH Annex VIII, information requirement section 8.5, data on acute toxicity shall be provided for at least one other route in addition to acute oral toxicity, depending on the nature of the substance and the likely route of human exposure.  Data are provided on the toxicity of the test material via the dermal route, as this route is considered the most likely route of potential exposure, and therefore data on the inhalation toxicity of the substance has been waived.

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:
23 February 2016 to 11 March 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: CRL:(WI) rats
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 10 weeks
- Weight at study initiation: 191-243 days
- Fasting period before study: On the night before treatment, the animals were fasted. The food but not water was withheld during an overnight period. The food was returned 3 hours after the treatment.
- Housing: 3 animals / cage. Cage type: Type II. polypropylene/polycarbonate. Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: 19-22 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19.3 - 22.8 °C
- Humidity: 30-68 %
- Air changes: 15-20 air exchanges/hour
- Photoperiod: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Route of administration:
oral: gavage
Vehicle:
DMSO
Details on oral exposure:
VEHICLE
- DMSO Lot/batch no. (if required): SZBE2790V

DOSAGE PREPARATION:
- The powdered test material was freshly formulated at a concentration of 200 or 30 mg/mL in the vehicle on the day of administration. The formulation container was stirred continuously up to finishing the treatment.

CLASS METHOD
- The initial dose level was selected by the Study Director to be that which is most likely to produce mortality in some of the dosed animals. In the lack of any preliminary toxicological information, 2000 mg/kg bw was selected to be the starting dose.
- Initially, three female animals were treated with 2000 mg/kg bw of the test material. One animal died, therefore further 3 animals were treated at the dose level of 2000 mg/kg bw. As all three animals were found dead in this second dose group, further testing was needed at a dose level of 300 mg/kg bw.
- Three female animals were treated with 300 mg/kg bw of the test material and one animal died in this group, therefore further 3 animals were treated at the same dose level As only one animal died again in the 300 mg/kg bw group, no further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.
Doses:
300 and 2 000 mg/kg bw
No. of animals per sex per dose:
6 females per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter, where possible. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- The body weight was recorded on the day before treatment (Day -1), on the day of the treatment (Day 0) and weekly thereafter, where possible. Terminal body weights of animals found dead were recorded.
- Necropsy of survivors performed: Yes, macroscopic examination was performed on all animals. The surviving animals were sacrificed by exsanguination under pentobarbital anaesthesia. After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.
Statistics:
The method used was not intended to allow the calculation of a precise LD50 value. The test material was ranked into categories of Globally Harmonised Classification System (GHS (rev. 6) 2015). Clinical signs, body weight, body weight gain and gross macroscopic data were tabulated.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The test material caused mortality in four of 6 animals at a dose level of 2 000 mg/kg bw and in two of 6 animals at a dose level of 300 mg/kg bw.
Clinical signs:
- Decreased activity and hunched back were observed in all animals (12/12) following treatment with the test material.
- In the groups treated at a dose level of 2 000 mg/kg bw, incoordination (5/6), piloerection (5/6) and prone position (2/6) were also noticed.
- In the groups treated at a dose level of 300 mg/kg bw, incoordination (6/6), piloerection (4/6) and prone position (1/6) were recorded besides the decreased activity and hunched back symptoms. One animal also had decreased respiratory rate just before death.
Body weight:
- The animals that survived the treatment at a dose level of 2 000 mg/kg bw lost weight during the first week, but gained weight during the second week. The surviving animals of the two groups of 300 mg/kg bw treatment gained weight continuously during 14-days observation period.
Gross pathology:
- Reddish cheesy material in the digestive contents of the stomach mixed with diet was found in the rats that died during the observation period; this was considered to be the administered test material.
- No macroscopic findings were noted in the surviving animals dosed at 300 mg/kg bw or 2 000 mg/kg bw and terminated on Day 14.

Table 1: Clinical Observations

Dose Level (mg/kg bw)

Animal No.

Observations

Observation Days

0

1

2

3

4

5

6

7-14

Frequency

30 min

1h

2h

3h

4h

6h

2000

8199

Symptom Free

-

-

-

-

-

-

-

-

+

+

+

+

+

12/20

Activity decreased

1

1

2

2

2

2

1

-

-

-

-

-

-

7/20

Hunched back

+

+

+

+

+

+

+

+

-

-

-

-

-

8/20

Incoordination

-

1

1

1

-

-

-

-

-

-

-

-

-

3/20

Piloerection

-

-

+

+

+

+

+

+

-

-

-

-

-

6/20

8200

Activity decreased

1

1

2

2

2

2

 

 

 

 

 

 

 

6/6

Hunched back

+

+

+

+

+

+

 

 

 

 

 

 

 

6/6

Incoordination

-

2

1

1

-

-

 

 

 

 

 

 

 

3/6

Piloerection

-

-

+

+

+

+

 

 

 

 

 

 

 

4/6

Found Dead

-

-

-

-

-

-

+

 

 

 

 

 

 

-

8201

Symptom Free

-

-

-

-

-

-

+

+

+

+

+

+

+

14/20

Activity decreased

1

1

2

1

-

-

-

-

-

-

-

-

-

4/20

Hunched back

+

+

+

+

+

+

-

-

-

-

-

-

-

6/20

Incoordination

-

-

1

1

-

-

-

-

-

-

-

-

-

2/20

Piloerection

-

-

+

+

+

+

-

-

-

-

-

-

-

4/20

8202

Activity decreased

1

2

 

 

 

 

 

 

 

 

 

 

 

2/2

Hunched back

+

-

 

 

 

 

 

 

 

 

 

 

 

1/2

Prone Position

-

+

 

 

 

 

 

 

 

 

 

 

 

1/2

Found Dead

-

-

+

 

 

 

 

 

 

 

 

 

 

-

8203

Activity decreased

1

2

2

2

2

2

 

 

 

 

 

 

 

6/6

Hunched back

+

+

+

+

+

+

 

 

 

 

 

 

 

6/6

Incoordination

-

1

2

1

1

1

 

 

 

 

 

 

 

5/6

Piloerection

-

-

+

+

+

+

 

 

 

 

 

 

 

4/6

Found Dead

-

-

-

-

-

-

+

 

 

 

 

 

 

-

8204

Activity decreased

1

2

2

2

2

3

 

 

 

 

 

 

 

6/6

Hunched back

+

+

+

+

+

-

 

 

 

 

 

 

 

5/6

Prone Position

-

-

-

-

-

+

 

 

 

 

 

 

 

1/6

Incoordination

1

1

1

1

1

-

 

 

 

 

 

 

 

5/6

Piloerection

-

-

+

+

+

+

 

 

 

 

 

 

 

4-6

Found Dead

-

-

-

-

-

-

+

 

 

 

 

 

 

-

300

8205

Activity decreased

1

1

1

1

2

3

 

 

 

 

 

 

 

6/6

Hunched back

+

+

+

+

+

-

 

 

 

 

 

 

 

5/6

Prone Position

-

-

-

-

-

+

 

 

 

 

 

 

 

1/6

Incoordination

-

1

1

1

1

-

 

 

 

 

 

 

 

4/6

Piloerection

-

-

+

+

+

+

 

 

 

 

 

 

 

4/6

Respiratory Rate Decreased

-

-

-

-

-

2

 

 

 

 

 

 

 

1/6

Found Dead

-

-

-

-

-

-

+

 

 

 

 

 

 

-

8206

Symptom Free

-

-

-

-

-

-

+

+

+

+

+

+

+

14/20

Activity Decreased

1

1

1

1

-

-

-

-

-

-

-

-

-

4/20

Hunched back

+

+

+

+

+

+

-

-

-

-

-

-

-

6/20

Incoordination

-

1

-

-

-

-

-

-

-

-

-

-

-

1/20

Piloerection

-

-

-

+

+

-

-

-

-

-

-

-

-

2/20

8207

Symptom Free

-

-

-

-

-

-

+

+

+

+

+

+

+

14/20

Activity Decreased

1

1

1

1

-

-

-

-

-

-

-

-

-

4/20

Hunched back

+

+

+

+

+

+

-

-

-

-

-

-

-

6/20

Incoordination

-

1

-

-

-

-

-

-

-

-

-

-

-

1/20

8208

Symptom Free

-

-

-

-

-

+

+

+

+

+

+

+

+

15/20

Activity Decreased

1

1

1

-

-

-

-

-

-

-

-

-

-

3/20

Hunched back

+

+

+

+

+

-

-

-

-

-

-

-

-

5/20

Incoordination

-

1

-

-

-

-

-

-

-

-

-

-

-

1/20

8209

Activity decreased

1

1

1

1

2

1

 

 

 

 

 

 

 

6/6

Hunched back

+

+

+

+

+

+

 

 

 

 

 

 

 

6/6

Incoordination

-

1

1

1

1

1

 

 

 

 

 

 

 

5/6

Piloerection

-

-

-

+

+

+

 

 

 

 

 

 

 

3/6

Found Dead

-

-

-

-

-

-

+

 

 

 

 

 

 

-

8210

Symptom Free

-

-

-

-

-

-

+

+

+

+

+

+

+

14/20

Activity Decreased

1

1

1

1

-

-

-

-

-

-

-

-

-

4/20

Hunched back

+

+

+

+

+

+

-

-

-

-

-

-

-

6/20

Incoordination

-

1

1

-

-

-

-

-

-

-

-

-

-

2/20

Piloerection

-

-

-

+

+

-

-

-

-

-

-

-

-

2/20

 

+ = present, - = absent, Frequency of observation = number of occurrence of observation / total number of observations, Severities: 1 = Slight/Small/Few; 2 = Moderate/Medium; 3 = Marked/Large/Many

Interpretation of results:
other: EU Criteria: Category 4: Harmful if swallowed (H302).
Conclusions:
Under the conditions of this study, the acute oral LD50 value of the test material was found to be between 300 and 2 000 mg/kg bw in female CRL:(WI) rats.
Executive summary:

The single-dose oral toxicity of the test material was investigated in accordance with the standardised guidelines OECD 423, EU Method B.1.Tris and OPPTS 870.1100, under GLP conditions using the acute toxic class method in CRL:(WI) rats.

Two groups of three female CRL:(WI) rats were treated with the test material at a dose level of 2 000 mg/kg bw (Group 1 and Group 2) and two groups of three female CRL:(WI) rats at a dose level of 300 mg/kg bw (Group 3 and Group 4).

A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test material was administered formulated in DMSO at a concentration of 200 or 30 mg/mL at a dose volume of 10 mL/kg bw.

Initially, three females (Group 1) were treated at a dose level of 2 000 mg/kg bw. As only one animal was found dead, a confirmatory group (Group 2) was treated at the same dose level. All three animals died in this confirmatory group, therefore the next dose level was 300 mg/kg bw. As only one animal was found dead in the group treated at a dose level of 300 mg/kg bw (Group 3), a confirmatory group (Group 4) was treated at the same dose level. In this group also only one animal died; therefore no further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter, where possible. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

The test material caused mortality in four of 6 animals at a dose level of 2 000 mg/kg bw and in two of 6 animals at a dose level of 300 mg/kg bw.

Decreased activity and hunched back were observed in all animals (12/12) following treatment with the test material. In the groups treated at a dose level of 2 000 mg/kg bw, incoordination (5/6), piloerection (5/6) and prone position (2/6) were also noticed. In the groups treated at a dose level of 300 mg/kg bw, incoordination (6/6), piloerection (4/6) and prone position (1/6) were recorded besides the decreased activity and hunched back symptoms. One animal also had decreased respiratory rate just before death.

The animals that survived the treatment at a dose level of 2 000 mg/kg bw lost weight during the first week, but gained weight during the second week. The surviving animals of the two groups of 300 mg/kg bw treatment gained weight continuously during 14-days observation period.

Reddish cheesy material in the digestive contents of the stomach mixed with diet was found in the rats that died during the observation period; this was considered to be the administered test material. No macroscopic findings were noted in the surviving animals dosed at 300 mg/kg bw or 2 000 mg/kg bw and terminated on Day 14.

Under the conditions of this study, the acute oral LD50 value of the test material was found to be between 300 and 2 000 mg/kg bw in female CRL:(WI) rats.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Quality of whole database:
The study was conducted according to standardised guidelines, in compliance with GLP, and as such was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
In accordance with Column 2 of REACH Annex VIII, information requirement section 8.5, data on acute toxicity shall be provided for at least one other route in addition to acute oral toxicity, depending on the nature of the substance and the likely route of human exposure. Data are provided on the toxicity of the test material via the dermal route, as this route is considered the most likely route of potential exposure based on the form of the substance inported and used in the EU. Therefore data on the inhalation toxicity of the substance has been waived.
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:
18 February 2019 to 06 March 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
09 October 2017
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
30 May 2008.
The guideline has not yet been revised in line with the OECD 2017 version. The study does not fully comply with the older version of the guideline, but the study design is considered to be acceptable for all OECD countries.
Deviations:
not applicable
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
August 1998.
The guideline has not yet been revised in line with the OECD 2017 version. The study does not fully comply with the older version of the guideline, but the study design is considered to be acceptable for all OECD countries.
Deviations:
not applicable
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
FORM AS APPLIED IN THE TEST
- The test material was powdered into fine dust before the application. Sufficient water was used to moisten the test material to ensure good contact with the skin.
- Measurement of pH: If the pH is 2 or less or 11.5 or greater, a study cannot be conducted, unless there is evidence that the test material is not severely irritating or corrosive to the skin. The pH of the test material in this study was determined prior to the initiation of the experiment and it was found to be 5.36, therefore the experiment could be started.
Species:
rat
Strain:
Wistar
Remarks:
Crl: WI Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 11 weeks
- Weight at study initiation: 244 - 256 g
- Housing: Animals were housed in type II polypropylene/polycarbonate cages. Animals were housed individually during the treatment and were group housed after patch removal. Rodents were housed with deep wood sawdust bedding to allow digging and other normal rodent activities. Lignocel 3/4-S Hygienic Animal Bedding was available to animals during the study. The quality of the bedding was guaranteed by the supplier. Additionally, nest building material (Arbocel Crinklets natural) was available to animals during the study. The quality of the nest building material was guaranteed by the supplier.
- Diet: Ad libitum
- Water: Tap water ad libitum.
- Acclimation period: 25 or 27 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20.2 - 24.5 °C
- Humidity: 25 - 73 %
- Air changes: 15 - 20 air exchanges/hour
- Photoperiod: 12 hours light (from 6.00 a.m. to 6.00 p.m.) / 12 hours dark, daily.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Remarks:
Sufficient water was used to moisten the test material to ensure good contact with the skin.
Details on dermal exposure:
TEST SITE
- Area of exposure: The back of each animal was shaved approximately 24 hours prior to treatment. The test material was applied to the shaved skin as a single dose and remained in contact with the skin for the exposure period.
- % coverage: approximately 10 % area of the total body surface.
- Type of wrap if used: Sterile gauze pads were placed on the skin of rats to cover the test material. These gauze pads were kept in contact with the skin using a patch with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing: At the end of the exposure period, the treated area of skin with the test material was washed with water at body temperature.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied: 0.50 ± 0.01 g
- For solids, paste formed: Sufficient water was used to moisten the test material to ensure good contact with the skin.
Duration of exposure:
24 hours
Doses:
2 000 mg/kg bw
No. of animals per sex per dose:
3 females
Initially one animal was dosed at the selected limit dose (2 000 mg/kg bw). As the animal survived, the second and third animal received the same dose.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/morbidity was checked twice daily during the 14-day observation period. Clinical observations were performed on the day of treatment at approximately 30 minutes, 1, 2 and 5 hours after application of the test material and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.
The body weights were recorded on Day 0 (before the test material administration) and on Days 7 and 14 (before necropsy).
- Necropsy of survivors performed: Yes. Macroscopic examination was performed on all animals. All animals were anaesthetised with sodium pentobarbital and exsanguinated. Following confirmation of death, after examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.
Statistics:
The method used was not intended to allow the calculation of a precise LD50 value.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
The test material did not cause mortality at the dose level of 2 000 mg/kg bw.
Clinical signs:
There were no adverse clinical signs noted in any animals during the 14-day observation period.
No adverse local dermal signs (erythema/oedema) were noted in any animals during the 14-day observation period.
Coloured skin (orange, on the back) was noted for all animals from Day 1 up to Day 5, which was related to the test material and was not identified as a local dermal sign. Test material residue on the back of the animals (in the treated area, not removable) was noted from Day 1 up to Day 9.
Body weight:
Slight decrease in body weight was observed in the animals of the main phase between Day 0 and Day 7. Body weights were within the range commonly recorded for this strain and age between Day 7 and Day 14 in case of all animals.
Gross pathology:
There was no evidence of any gross macroscopic changes at a dose level of 2 000 mg/kg bw.
Interpretation of results:
other: Not classified according to EU criteria
Conclusions:
Under the conditions of the study the acute dermal LD50 of the test material was determined to be greater than 2 000 mg/kg body weight in female rats.
Executive summary:

The acute dermal toxicity of the test material was investigated in a study which was conducted in compliance with the standardised guideline OECD 402, under GLP conditions.

A single animal at a dose level of 2 000 mg/kg body weight (bw) was used in a range-finding phase, followed by two animals in the main phase to confirm the expected non-lethal dose level. The test material was applied as a single dermal 24-hour exposure followed by a 14-day observation period.

Clinical observations were performed on all animals at approximately 30 minutes, 1, 2, 5 hours after dosing and daily for 14 days thereafter. Body weight was measured on Day 0 (prior to dosing) and on Days 7 and 14 (before necropsy). Gross macroscopic examination was performed on all animals at necropsy at the end of the 2-week observation period (Day 14).

The test material did not cause mortality at the dose level of 2 000 mg/kg bw.

There were no adverse clinical signs noted in any animals during the 14-day observation period.

No adverse local dermal signs (erythema/oedema) were noted in any animals during the 14-day observation period.

Coloured skin (orange, on the back) was noted for all animals from Day 1 up to Day 5, which was related to the test material and was not identified as a local dermal sign. Test material residue on the back of the animals (in the treated area, not removable) was noted from Day 1 up to Day 9.

Slight decrease in body weight was observed in the animals of the main phase between Day 0 and Day 7. Body weights were within the range commonly recorded for this strain and age between Day 7 and Day 14 in case of all animals.

Additionally, there was no evidence of any macroscopic changes at a dose level of 2 000 mg/kg bw.

Under the conditions of the study the acute dermal LD50 of the test material was determined to be greater than 2 000 mg/kg body weight in female rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Acute Oral Toxicity

The single-dose oral toxicity of the test material was investigated in accordance with the standardised guidelines OECD 423, EU Method B.1.Tris and OPPTS 870.1100, under GLP conditions using the acute toxic class method in CRL:(WI) rats. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Two groups of three female CRL:(WI) rats were treated with the test material at a dose level of 2 000 mg/kg bw (Group 1 and Group 2) and two groups of three female CRL:(WI) rats at a dose level of 300 mg/kg bw (Group 3 and Group 4). A single oral treatment was carried out by gavage for each animal after an overnight food withdrawal. Food was made available again 3 hours after the treatment. The test material was administered formulated in DMSO at a concentration of 200 or 30 mg/mL at a dose volume of 10 mL/kg bw.

Initially, three females (Group 1) were treated at a dose level of 2 000 mg/kg bw. As only one animal was found dead, a confirmatory group (Group 2) was treated at the same dose level. All three animals died in this confirmatory group, therefore the next dose level was 300 mg/kg bw. As only one animal was found dead in the group treated at a dose level of 300 mg/kg bw (Group 3), a confirmatory group (Group 4) was treated at the same dose level. In this group also only one animal died; therefore no further testing was required according to OECD 423 and Commission Regulation (EC) NO 440/2008 of 30 May 2008, B.1.Tris.

Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter, where possible. Body weight was measured on Days -1, 0 and 7 and before necropsy. All animals were subjected to a necropsy and a macroscopic examination.

The test material caused mortality in four of 6 animals at a dose level of 2 000 mg/kg bw and in two of 6 animals at a dose level of 300 mg/kg bw. Decreased activity and hunched back were observed in all animals (12/12) following treatment with the test material. In the groups treated at a dose level of 2 000 mg/kg bw, incoordination (5/6), piloerection (5/6) and prone position (2/6) were also noticed. In the groups treated at a dose level of 300 mg/kg bw, incoordination (6/6), piloerection (4/6) and prone position (1/6) were recorded besides the decreased activity and hunched back symptoms. One animal also had decreased respiratory rate just before death.

The animals that survived the treatment at a dose level of 2 000 mg/kg bw lost weight during the first week, but gained weight during the second week. The surviving animals of the two groups of 300 mg/kg bw treatment gained weight continuously during 14-days observation period.

Reddish cheesy material in the digestive contents of the stomach mixed with diet was found in the rats that died during the observation period; this was considered to be the administered test material. No macroscopic findings were noted in the surviving animals dosed at 300 mg/kg bw or 2 000 mg/kg bw and terminated on Day 14.

Under the conditions of this study, the acute oral LD50 value of the test material was found to be between 300 and 2 000 mg/kg bw in female CRL:(WI) rats.

Acute Dermal Toxicity

The acute dermal toxicity of the test material was investigated in a study which was conducted in compliance with the standardised guideline OECD 402, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

A single animal at a dose level of 2 000 mg/kg body weight (bw) was used in a range-finding phase, followed by two animals in the main phase to confirm the expected non-lethal dose level. The test material was applied as a single dermal 24-hour exposure followed by a 14-day observation period.

Clinical observations were performed on all animals at approximately 30 minutes, 1, 2, 5 hours after dosing and daily for 14 days thereafter. Body weight was measured on Day 0 (prior to dosing) and on Days 7 and 14 (before necropsy). Gross macroscopic examination was performed on all animals at necropsy at the end of the 2-week observation period (Day 14).

The test material did not cause mortality at the dose level of 2 000 mg/kg bw.

There were no adverse clinical signs noted in any animals during the 14-day observation period.

No adverse local dermal signs (erythema/oedema) were noted in any animals during the 14-day observation period.

Coloured skin (orange, on the back) was noted for all animals from Day 1 up to Day 5, which was related to the test material and was not identified as a local dermal sign. Test material residue on the back of the animals (in the treated area, not removable) was noted from Day 1 up to Day 9.

Slight decrease in body weight was observed in the animals of the main phase between Day 0 and Day 7. Body weights were within the range commonly recorded for this strain and age between Day 7 and Day 14 in case of all animals.

Additionally, there was no evidence of any macroscopic changes at a dose level of 2 000 mg/kg bw.

Under the conditions of the study the acute dermal LD50 of the test material was determined to be greater than 2 000 mg/kg body weight in female rats.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance is classified as Category 4: Harmful if swallowed (H302).

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