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Administrative data

Description of key information

Oral: LD50>2000 mg/kg bw, female, rat, OECD 425, Sieber 2011
Dermal: LD50 >2000 mg/kg bw, male/female, rat, OECD 402, Sieber 2011

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:
from 2011-04-19 to 2011-05-19 (experimental phase)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Version / remarks:
from 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
from December 2002
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
up-and-down procedure
Limit test:
no
Species:
rat
Strain:
other: RccHan:WIST(SPF)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 10 weeks
- Weight at study initiation: 181.8 - 185.7 g
- Fasting period before study: 17 to 22 hours before treatment, with free access to water
- Housing: In groups of up to five in Makrolon type-4 cages during acclimatization. Individually in Makrolon type-3 cages during treatment and observation. Cages were equipped with wire mesh tops and standard softwood bedding including paper enrichment during treatment and observation.
- Diet (e.g. ad libitum): Pelleted rodent maintenance diet ad libitum, except for the overnight fasting period
- Water (e.g. ad libitum): Community tap water ad libitum
- Acclimation period: At least 5 days under laboratory conditions, after health examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 55 and 200 mg/mL
- Amount of vehicle (if gavage):
94.5 mg/kg CMC (55 mg/mL preparation)
80 mg/kg CMC (200 mg/mL preparation)
Calculation: amount CMC in 10 mL preparation= 1% · (10000 mg preparation - x mg substance per 10000 mg preparation), where x is 550 and 2000 mg, respectively
- Justification for choice of vehicle: The vehicle was chosen after non-GLP solubility testing. Purified water and 0.5% CMC in purified water were found unsuitable vehicles for oral gavage application, as the test item could not be sufficiently dispersed in either vehicle. By grinding the test item in a mortar and pestle together with 1% CMC in purified water, a 20% (weight:weight) dispersion of the test item suitable for oral gavage application was obtained.
- Lot/batch no. (if required): 0001414646
- Purity: not reported

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

DOSAGE PREPARATION (if unusual): Dose formulations were prepared shortly before each treatment using a magnetic stirrer and a spatula as homogenizers. The test item was weighed on a suitable precision balance and supplied to a mortar. By using a pestle, the test item and almost all of the vehicle was ground to a dispersion. This dispersion was transferred to a volumetric beaker and the vehicle was added up to the defined volume (weight:volume). Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.
Doses:
550 mg/kg bw and 2000 mg/kg bw
No. of animals per sex per dose:
1 female with 550 mg/kg bw
3 females with 2000 mg/kg bw
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were examined daily during acclimatization and mortality and clinical signs were assessed. All animals were examined for clinical signs before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2 to 15. Mortality was assessed before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (with the clinical signs) and twice daily during test days 2 to 15. Body weights were recorded on the last day of acclimatization (prior to removal of food), on test day 1 (prior to treatment) and on test days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: mortality, clinical signs, body weight, macroscopy
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred during the course of the study.
Clinical signs:
One animal treated with 550 mg/kg (animal no. 1) and two of the three animals treated with 2000 mg/kg (animals no. 2 and no. 4) showed ruffled fur after treatment on test day 1. In one animal treated with 2000 mg/kg (animal no. 2), ruffled fur was still observed on test day 2. No clinical signs were observed in any animal during acclimatization and from test day 2 until the end of the observation period.
Body weight:
The body weights were 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:
The median lethal dose (LD50) of the test item after single oral administration to female rats, observed over a period of 14 days, is >2000 mg/kg body weight. The study is considered to be adequate and reliable.
Executive summary:

An acute oral toxicity test with 4 animals (female RccHan:WIST(SPF) rat) was conducted under GLP according to OECD TG 425.

The animals were treated with the test item by gavage at doses of 550 and 2000 mg/kg body weight. Dosing started in one female animal at a dose level of 550 mg/kg. As no death occurred and no severe clinical signs were observed at this dose level, the next female was treated at a dose level of 2000 mg/kg. Two more females were treated at a dose level of 2000 mg/kg, as at this dose no deaths occurred and no severe clinical signs were observed.

All animals were examined for clinical signs before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2 to 15. Mortality was assessed before treatment, once within the first

30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (with the clinical signs) and twice daily during test days 2 to 15. Body weights were recorded on the last day of acclimatization (prior to removal of food), on test day 1 (prior to treatment) and

on test days 8 and 15. All animals were examined macroscopically after being sacrificed at the end of the observation period.

No deaths occurred during the course of the study. The animal treated with 550 mg/kg and two of the three animals treated with 2000 mg/kg showed ruffled fur after treatment on test day 1. In one animal treated with 2000 mg/kg, ruffled fur was still observed on test day 2. No clinical signs were observed in any animal during acclimatization and from test day 2 until the end of the observation period. The body weights were within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

The median lethal dose (LD50) of the test item after single oral administration to female rats, observed over a period of 14 days, is >2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
2 000 mg/kg bw
Quality of whole database:
GLP compliant, OECD 425

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:
from 2011-04-19 to 2011-05-12 (experimental phase)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
adopted February 24, 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
from May 30, 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
from August 1998
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: RccHan: WIST(SPF)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: males: 9 weeks, females: 11 weeks
- Weight at study initiation: males: 250.4 g - 260.8 g, females: 190.8 g - 208.2 g
- Fasting period before study: No
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding, including paper enrichment. Individually in Makrolon type-3 cages and under similar conditions during treatment and observation.
- Diet (e.g. ad libitum): Pelleted Teklad Rat-Mouse Diet 2914C ad libitum
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: At least 5 days, under laboratory conditions and after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To:
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: backs of the animals
- Type of wrap if used: The patch was covered with a semi-occlusive dressing which was wrapped around the abdomen and fixed with an elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed with lukewarm tap water and dried with disposable paper towels
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg body weight
- Constant volume or concentration used: not applicable (only 1 concentration)
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): The test item was moistened with a minimum of purified water to obtain a dry paste in order to guarantee good skin contact.
Duration of exposure:
24 hours
Doses:
2000 mg/kg body weight (applied by weight as supplied by the Sponsor)
No. of animals per sex per dose:
group 1: 1 male, 1 female (Animal no. 1 and 2, treated 2 days before the other rats)
group 2: 4 males, 4 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were examined daily during acclimatization (mortality and clinical signs). All animals were examined for clinical signs before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2 to 15. Local dermal signs were assessed once daily from test day 2 to 15. Mortality was assessed before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (with the clinical signs) and twice daily during days 2 to15. Body weights were recorded on test day 1 (prior to administration) and on test days 8 and 15. All animals were examined macroscopically after being sacrificed at the end of the study.
- Necropsy of survivors performed: yes
- Other examinations performed: mortality, clinical signs, local dermal signs, body weight, macroscopy
Preliminary study:
2 rats (1 male (animal no. 1), 1 female (animal no. 2)) were treated 2 days before the other rats. The findings for these were:

- no deaths, no clinical signs, no macroscopic findings
- body weights within commonly recorded range
- focal crusts after test item application lasting for 13 days in male rat
- Brown discoloration of the skin in male and female rat lasting up to the end of the observation period
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred during the course of the study.
Clinical signs:
No clinical signs were observed during the course of the study.
Body weight:
One female slightly lost body weight in the second week after treatment (2%). The body weights of the other animals were within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necropsy.
Other findings:
Local dermal signs: Four males and one female showed focal crusts after test item application lasting for a minimum of 2 and a maximum of 13 days. Brown discoloration of the skin was observed in all animals for a minimum of 7 days after test item application and lasted up to the end of the observation period in 3 males and 3 females.

Local Dermal Signs

Animal no/sex

Dose [mg/kg]

Local dermal signs (max grade)

Test day

2

3

4

5

6

7

8

9

10

11

12

13

14

15

1 / M

2000

Focal crusts

1

1

1

1

1

1

1

1

1

1

1

1

1

.

Brown discolouration

2

2

2

2

2

2

2

2

2

1

1

1

1

1

2 / F

2000

Brown discolouration

2

2

2

2

2

2

2

1

1

1

1

1

1

1

3/ M

2000

Focal crusts

.

.

.

.

.

1

1

1

1

1

.

.

.

.

Brown discolouration

2

2

2

2

2

1

1

1

1

1

1

1

1

1

4 / M

2000

Focal crusts

1

1

1

1

1

1

1

.

.

.

.

.

.

.

Brown discolouration

2

2

2

2

2

1

1

1

1

1

1

1

1

1

5 / M

2000

Focal crusts

.

.

.

.

.

1

1

.

.

.

.

.

.

.

Brown discolouration

2

2

2

2

2

1

1

1

1

1

.

.

.

.

6 / M

2000

Brown discolouration

2

2

2

2

2

1

1

.

.

.

.

.

.

.

7 / F

2000

Brown discolouration

1

1

1

1

1

1

1

1

1

1

1

1

1

.

8 / F

2000

Brown discolouration

2

2

2

2

2

2

2

2

2

2

2

2

2

2

9 / F

2000

Focal crusts

.

.

.

.

.

1

1

.

.

.

.

.

.

.

Brown discolouration

2

2

2

2

2

2

1

1

1

1

1

1

1

1

10 / F

2000

Brown discolouration

1

1

1

1

1

1

1

1

1

1

.

.

.

.

 

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The median lethal dose (LD50) of the test item after single dermal administration to rats of both sexes, observed over a period of 14 days, is greater than 2000 mg/kg body weight. The study is considered to be adequate and reliable.
Executive summary:

Acute dermal toxicity was assessed in a valid GLP study according to OECD TG 402.

5 male and 5 female RccHan:WIST (SPF) rats were treated with the test item at a dose level of 2000 mg/kg by dermal application. The test item was applied by weight as supplied by the Sponsor. For better dermal contact, the test item was moistened with purified water before application. All animals were examined for clinical signs before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2 to 15. Local dermal signs were assessed once daily from test day 2 to 15. Mortality was assessed before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (with the clinical signs) and twice daily during test days 2 to15. Body weights were recorded on test day 1 (prior to administration) and on test days 8 and 15. All animals were examined macroscopically after being sacrificed at the end of the study.

No deaths occurred during the course of the study and no clinical signs were observed. Four males and one female showed focal crusts after test item application lasting for a minimum of 2 and a maximum of 13 days. Brown discoloration of the skin was observed in all animals for a minimum of 7 days after test item application and lasted up to the end of the observation period in 3 males and 3 females. One female slightly lost body weight in the second week after treatment (2%). The body weights of the other animals were within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

The median lethal dose of the test item after single dermal administration to rats of both sexes, observed over a period of 14 days, is greater than 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
GLP compliant, OECD 402

Additional information

Acute oral toxicity

An acute oral toxicity test with 4 animals (female RccHan:WIST(SPF) rat) was conducted under GLP according to OECD TG 425.

The animals were treated with the test item by gavage at doses of 550 and 2000 mg/kg body weight. Dosing started in one female animal at a dose level of 550 mg/kg. As no death occurred and no severe clinical signs were observed at this dose level, the next female was treated at a dose level of 2000 mg/kg. Two more females were treated at a dose level of 2000 mg/kg, as at this dose no deaths occurred and no severe clinical signs were observed.

All animals were examined for clinical signs before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2 to 15. Mortality was assessed before treatment, once within the first

30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (with the clinical signs) and twice daily during test days 2 to 15. Body weights were recorded on the last day of acclimatization (prior to removal of food), on test day 1 (prior to treatment) and on test days 8 and 15. All animals were examined macroscopically after being sacrificed at the end of the observation period.

Acute dermal toxicity

An acute dermal toxicity was assessed in a valid GLP study according to OECD TG 402.

5 male and 5 female RccHan:WIST (SPF) rats were treated with the test item at a dose level of 2000 mg/kg by dermal application. The test item was applied by weight as supplied by the Sponsor. For better dermal contact, the test item was moistened with purified water before application. All animals were examined for clinical signs before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2 to 15. Local dermal signs were assessed once daily from test day 2 to 15. Mortality was assessed before treatment, once within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (with the clinical signs) and twice daily during test days 2 to15. Body weights were recorded on test day 1 (prior to administration) and on test days 8 and 15. All animals were examined macroscopically after being sacrificed at the end of the study.

No deaths occurred during the course of the study and no clinical signs were observed. Four males and one female showed focal crusts after test item application lasting for a minimum of 2 and a maximum of 13 days. Brown discoloration of the skin was observed in all animals for a minimum of 7 days after test item application and lasted up to the end of the observation period in 3 males and 3 females. One female slightly lost body weight in the second week after treatment (2%). The body weights of the other animals were within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

The median lethal dose of the test item after single dermal administration to rats of both sexes, observed over a period of 14 days, is greater than 2000 mg/kg body weight.

Justification for classification or non-classification

Acute oral toxicity

The acute oral toxicity of the substance is > 2000 mg/kg. As a result it does not meet the criteria for classification according to Regulation (EC) No. 1272/2008, Annex I, Part 3, 3.1.2.

 

Acute dermal toxicity

The acute dermal toxicity of the substance is > 2000 mg/kg. As a result it does not meet the criteria for classification according to Regulation (EC) No. 1272/2008, Annex I, Part 3, 3.1.2.