Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Physical state: Brown powder
- Stability under test conditions: not reported
- Storage condition of test material: At room temperature (20 ± 5 °C), light protected

Test animals

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:

Administration / exposure

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

Results and discussion

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
Effect levels
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.

Any other information on results incl. tables

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

.

.

.

.

 

Applicant's summary and conclusion

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.