Registration Dossier

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Administrative data

Description of key information

Acute oral toxicity, OECD 423, van Huygevoort (2020): LD50: > 2000 mg/kg bw


Acute dermal toxicity, OECD 402, van Huygevoort (2020): LD50: > 2000 mg/kg bw

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:
27 March 2020 - 07 May 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Test conducted in accordance with OECD Test Guideline and in accordance with GLP. All validity criteria were met.
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:
EC No 440/2008, part B
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMAFF Guidelines
Version / remarks:
2000, including the most recent revisions.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Remarks:
Crl: WI(Han). Condition: Outbred, SPF-Quality
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult animals (approximately 8-10 weeks old) were selected
- Weight at study initiation: 143 to 178 g
- Fasting period before study: Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the test item. Water was available.
- Housing: On arrival and following assignment to the study, animals were group housed (up to 3 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized wooden fibers as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room in which the animals were kept was documented in the study records.
Animals were separated during designated procedures/activities. Each cage was clearly labeled.
- Historical data: Not provided
- Diet: ad libitum
Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water: ad libitum
Municipal tap-water was freely available to each animal via water bottles.
Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period: The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.
- Microbiological status when known: Not reported
- Method of randomisation in assigning animals to test and control groups: Not reported - animals were assigned to the study at the discretion of the coordinating biotechnician, with all animals within ± 20 % of the sex mean body weights. Animals in poor health or at extremes of body weight range were not assigned to the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Daily mean temperature during the study period was 20 to 21°C
- Humidity (%): Daily mean relative humidity of 43 to 52 %
- Air changes (per hr): => 10, with 100 % fresh air (no recirculation)
- Photoperiod (hrs dark / hrs light): 12 h light / 12 h dark

IN-LIFE DATES: From: 27 March 2020 To: 07 May 2020
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
VEHICLE
N/A

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg body weight (bw)

DOSAGE PREPARATION (if unusual): The test item was dosed as received.
A single dose of test item was administered to the appropriate animals by oral gavage on Day 1, using a syringe with a plastic gavage cannula attached.
The dose volume for each animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows: Dose level (g/kg) / spec.gravity.
The dosing formulations were stirred continuously during dose administration protected from light.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
The dose levels were based on the OECD test guidelines and were selected from the series 5 (lowest dose level), 50, 300 and 2000 (highest dose level) mg/kg bw. The starting dose level should be the one that is likely to produce mortality in at least some of the animals and was selected based on available toxicity data of the test item.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
3 females at 300 mg/kg bw
6 females (in 2 groups) at 2000 mg/mg bw
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Post-dose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.
Animals were weighed individually on Day 1 (pre-dose), 8 and 15. A fasted weight was recorded on the day of dosing.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes
- Other examinations performed: Maroscopic abnormalities.

Post-dose observations:
All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate). Signs were graded for severity and the maximum grade was predefined at 3 or 4. Grades were coded as slight (grade 1), moderate (grade 2), severe (grade 3) and very severe (grade 4). For certain signs, only its presence (grade 1) or absence (grade 0) was scored.
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).
All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented.
The oral LD50 value of the test item was ranked within the following ranges: 0-5, 5-50, 50- 300 or 300-2000 mg/kg b.w. or as exceeding 2000 mg/kg bw. The LD50 cut-off value was established based on OECD guideline 423.
Preliminary study:
N/A
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None
Clinical signs:
At 300 mg/kg, hunched posture and erected fur were noted for all animals on Day 1.
At 2000 mg/kg, hunched posture, erected fur, closed eyes, uncoordinated movements, decreased activity, shallow breathing and breathing with abnormal sounds were observed for the animals between Days 1 and 3. One animal showed a prostrate posture and was cold to the touch on Day 1.
Body weight:
The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
Gross pathology:
No abnormalities were found at macroscopic post-mortem examination of the animals.
Other findings:
The oral LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight.

Table 1. Individual mortality

Group

Dose Level (mg/kg bw)

Sex

Animal

Cage

Removal

Removal

Pathology

Day

Week

Date

Reason

1

300

Female

1

1

15

3

21Apr20

Terminal euthanasia

2

1

15

3

21Apr20

Terminal euthanasia

3

1

15

3

21Apr20

Terminal euthanasia

2

2000

Female

4

2

15

3

29Apr20

Terminal euthanasia

5

2

15

3

29Apr20

Terminal euthanasia

6

2

15

3

29Apr20

Terminal euthanasia

3

2000

Female

7

3

15

3

07May20

Terminal euthanasia

8

3

15

3

07May20

Terminal euthanasia

9

3

15

3

07May20

Terminal euthanasia

 

Table 2. Individual clinical observations

Group 1 Sex: Female 300 mg/kg

 

Observation Type: All Types

Day(s) Relative to Start Date

1
1 h

1
2 h

1
3 h

2

3

4

5

6

7

8

9

10

11

12

13

14

15

1

Hunched posture

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

2

Hunched posture

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

3

Hunched posture

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Group 2 Sex: Female 2000 mg/kg

 

Observation Type: All Types

Day(s) Relative to Start Date

1
1 h

1
2 h

1
3 h

2

3

4

5

6

7

8

9

10

11

12

13

14

15

4

Hunched posture

-

X

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

Eye closed, left, completely

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Eye closed, right, completely

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, slight

-

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

Activity decreased

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

5

Breathing, abnormal sounds

-

X

-

X

X

-

-

-

-

-

-

-

-

-

-

-

-

Breathing, shallow

-

-

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

Hunched posture

-

X

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

-

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, slight

-

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Group 3 Sex: Female 2000 mg/kg

 

Observation Type: All Types

Day(s) Relative to Start Date

1
1 h

1
2 h

1
3 h

2

3

4

5

6

7

8

9

10

11

12

13

14

15

7

Hunched posture

-

X

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, moderate

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, slight

-

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

8

Hunched posture

-

X

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, moderate

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, slight

-

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Activity decreased

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

9

Hunched posture

-

X

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

Fur, erected

X

X

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Eye closed, left, completely

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Eye closed, right, completely

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, moderate

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Uncoordinated, slight

-

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Activity decreased

-

X

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight.
Executive summary:

A study was performed in accordance with OECD 423 (2001), in order to determine the potential for acute oral toxicity of the test item.

The test item was administered in a single dose to female rats at two defined doses. Initially, the test item was administered by oral gavage to three female Wistar Han rats at 300 mg/kg bw. In a stepwise procedure two additional groups of three females were dosed at 2000 mg/kg bw. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred in any of the animals. At 300 mg/kg, hunched posture and erected fur were noted for all animals on Day 1. At 2000 mg/kg, hunched posture, erected fur, closed eyes, uncoordinated movements, decreased activity, shallow breathing and breathing with abnormal sounds were observed for the animals between Days 1 and 3. One animal showed a prostrate posture and was cold to the touch on Day 1. The body weight gain shown by the animals over the study period was considered to be normal. No abnormalities were found at macroscopic post-mortem examination of the animals.

The oral LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight. Based on the conditions of this test, the test item would not be classified for acute oral toxicity, in accordance with CLP Regulation No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Quality of whole database:
A Klimisch rating of 1 has been applied. The study is conducted to the relevant OECD test guideline and in accordance with GLP, therefore reliably fulfilling the endpoint requirement.

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:
14 August 2020 - 09 September 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Test conducted in accordance with OECD Test Guideline and in accordance with GLP
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)
Version / remarks:
09 October 2017
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Remarks:
Crl: WI(Han)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Rationale for use of males (if applicable): N/A
- Age at study initiation: Single young adult animal (approx. 10 weeks old)
- Weight at study initiation: 181 g
- Fasting period before study: Not specified
- Housing: On arrival, the animal was group housed (up to 5 animals of the same sex together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) and following assignment to the study, the animal was individually housed in polycarbonate cages (Makrolon MIII type; height 18 cm.) containing sterilized wooden fibers as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room in which the animal was kept was documented in the study records. The cage was clearly labeled.
- Historical data:
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum): Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.
- Acclimation period: The animal was allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.
- Microbiological status when known: Not reported
- Method of randomisation in assigning animals to test and control groups:
Animals were assigned to the study at the discretion of the coordinating biotechnician, with all animals within ± 20% of the sex mean body weights. Animals in poor health or at extremes of body weight range were not assigned to the study.
Before the initiation of dosing, a health inspection was performed, and any assigned animal considered unsuitable for use in the study were replaced by alternate animals obtained from the same shipment and maintained under the same environmental conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Actual daily mean temperature during study period: 21-22°C
- Humidity (%): Actual daily mean relative humidity during study period: 48 -70%
- Air changes (per hr): => 10/h with 100% fresh air (no air recirculation)
- Photoperiod (hrs dark / hrs light): 12 hour light/12 hour dark cycle
- Animal enrichment: For psychological/environmental enrichment, the animal was provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)

IN-LIFE DATES: From: 14 August 2020 To: 09 September 2020
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: The test item was applied to approx. 18 cm² .
- % coverage: Approx. 10 %
- Type of wrap if used: The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages.

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Constant volume or concentration used: yes/no
- For solids, paste formed: yes/no

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:
Duration of exposure:
Application period: 24 h, after which the dressing was removed, and the skin cleaned of residual test item using water.
Doses:
2000 mg/kg

The dose volume for the animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows: Dose level (g/kg) / spec.gravity or density (g/mL) * purity correction factor.

Animal welfare reasons prevented the initiation of the main study due to severe skin effects, therefore no addiitonal doses were applied.
No. of animals per sex per dose:
A single female was dosed at the single 2000 mg/kg bw dose.
Control animals:
other: Adjacent areas of untreated skin of the animal served as a control
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Throughout the study, the animal was observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Post-dose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The animal was examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate), particular attention being paid to the animal during and for the first hour after dosing.
The animal was weighed individually on Day 1 (pre-dose), 8 and 15.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes
- Other examinations performed: clinical signs, macroscopic findings
Statistics:
Not applicable, only a single dose was applied to a single animal during the test
Preliminary study:
The results from the single animal dosed at 2000 mg/kg are as follows:
No mortality occurred.
Systemic signs of toxicity consisted of hunched posture on Days 3-5.
Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15.
The body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study.
No abnormalities were found at macroscopic post-mortem examination of the animal.
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
Remarks:
Minor transient systemic toxicity (hunched posture) was observed and mortality is not to be expected if additional animals were dosed.
Mortality:
None
Clinical signs:
Minor transient systemic toxicity (hunched posture) was observed on Days 3-5.
Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15.
Body weight:
The body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study.
Gross pathology:
No abnormalities were found at macroscopic post-mortem examination of the animal.
Other findings:
N/A

Table 1. Individual Mortality - Female Day(s): - Relative to Start Date

2000

mg/kg Group 1

 

 

 

Day of Death

Removal Date

Path Removal Reason

1

15

09-Sep-2020

Terminal euthanasia

 

Table 2. Individual Clinical Observations

Group 1 Sex: Female 2000 mg/kg

 

Observation Type: All Types

Day(s) Relative to Start Date

1

0h

1

2h

1

4h

2

3

4

5

6

7

8

9

10

11

12

13

14

15

1

Hunched Posture

.

.

.

.

X

X

X

.

.

.

.

.

.

.

.

.

.

 

Skin, Flaking, Lumbar, Grade 1

.

.

.

.

.

X

X

X

.

.

.

.

.

X

.

X

X

 

Skin, Flaking, Lumbar, Grade 2

.

.

.

.

.

.

.

.

X

X

X

X

.

.

X

.

.

 

Skin, Flaking, Lumbar, Moderate

.

.

.

.

.

.

.

.

.

.

.

.

X

.

.

.

.

 

Skin, Discolored, Lumbar, Red

.

.

.

X

X

X

X

X

X

.

.

.

.

.

.

.

.

 

Skin, Scab, Lumbar

.

.

.

.

X

X

X

X

X

X

X

X

X

X

X

X

X

X = present

Table 3. Individual Body Weights - Female

2000

mg/kg Group 1

Day(s) Relative to Start Date

1

8

15

1

181

193

203

Mean

181.0

193.0

203.0

SD

-

-

-

N

1

1

1

 

Table 4. Irritation Observations

Group 1 Sex: Female 2000 mg/kg

 

Observation Type: All Types

Day(s) Relative to Start Date

3

24H

4

48H

5

72H

1

Erythema, Lumbar, Grade 1

X

X

.

 

Erythema, Lumbar, Grade 2

.

.

X

 

Other (see comment), Right Subcapsulara

X

X

X

a Scab on right flank showing necrosis.

X = Present

Individual Macroscopic Pathology:

Animal: 1, Group: 1, Sex: Female, Dose: 2000 mg/kg, Removal Reason: Terminal Euthanasia

Study Day (Week) of Death: 15 (3)

Gross Status: Complete

Gross Pathology Observations [Correlation]:

No observations found

Gross Pathology - The following Tissues were Not Examined:

None

 

Interpretation of results:
GHS criteria not met
Conclusions:
The dermal LD50 value of the test item in Wistar Han rats was established to be > 2000 mg/kg body weight.
Executive summary:

A test was performed in accordance with OECD 402 (2017), in order to determine the potential toxicity of the test item, when given by a single dermal dose.

Initially, the test item was administered to a single female Wistar Han rat by a single dermal application at 2000 mg/kg body weight for 24 hours in a range finder study. The animal was subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Furthermore, the body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study and no abnormalities were found at macroscopic post-mortem examination. Hunched posture was noted between Days 3 and 5. Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15. Based on the severity of these effects, it was considered that the other two animals assigned to the main study could not be dosed for animal welfare reasons.

Based on the results, it can be stated that the dermal LD50 value exceeds 2000 mg/kg, because only minor transient systemic toxicity (hunched posture) was observed and mortality is not to be expected if additional animals were dosed.

Based on these results, the test item would not be classified in accordance with Regulation (EC) No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Quality of whole database:
A Klimisch rating of 1 has been applied. The study is conducted to the relevant OECD test guideline and in accordance with GLP, therefore reliably fulfilling the endpoint requirement.

Additional information

Acute Oral toxicity:

A study was performed in accordance with OECD 423 (2001), in order to determine the potential for acute oral toxicity of the test item.

Initially, the test item was administered by oral gavage to three female Wistar Han rats at 300 mg/kg bw. In a stepwise procedure two additional groups of three females were dosed at 2000 mg/kg bw.

No mortality occurred in any of the animals. At 300 mg/kg, hunched posture and erected fur were noted for all animals on Day 1. At 2000 mg/kg, hunched posture, erected fur, closed eyes, uncoordinated movements, decreased activity, shallow breathing and breathing with abnormal sounds were observed for the animals between Days 1 and 3. One animal showed a prostrate posture and was cold to the touch on Day 1. The body weight gain shown by the animals over the study period was considered to be normal. No abnormalities were found at macroscopic post-mortem examination of the animals.

 

Acute Dermal toxicity:

A test was performed in accordance with OECD 402 (2017), in order to determine the potential toxicity of the test item, when given by a single dermal dose.

Initially, the test item was administered to a single female Wistar Han rat by a single dermal application at 2000 mg/kg body weight for 24 hours in a range finder study. The animal was subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. Furthermore, the body weight gain shown by the animal during the observation period was within the range expected for rats used in this type of study and no abnormalities were found at macroscopic post-mortem examination. Hunched posture was noted between Days 3 and 5. Local toxicity consisted of very slight to well-defined erythema between 24- and 72-hours post-dosing. Red discoloration was seen between Days 2 and 7, which resulted in flaking, signs of necrosis and/or scabs on the lumbar skin / right flank between Days 2 and 15. Based on the severity of these effects, it was considered that the other two animals assigned to the main study could not be dosed for animal welfare reasons.

Based on the results, it can be stated that the dermal LD50 value exceeds 2000 mg/kg, because only minor transient systemic toxicity (hunched posture) was observed and mortality is not to be expected if additional animals were dosed.

Justification for classification or non-classification

The oral LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight. Based on the conditions of this test, the test item would not be classified for acute oral toxicity, in accordance with CLP Regulation No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.

The dermal LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight. Based on the conditions of this test, the test item would not be classified for acute dermal toxicity, in accordance with CLP Regulation No 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.

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