Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
3 October 2012 to 17 October 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
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 (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
semi-solid (amorphous): gel

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: RccHan:WIST
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: The animals weighed at least 200 g. The bodyweight variation did not exceed ± 20 % of the mean weight for each sex.
- Fasting period before study: No.
- Housing: The animals were housed in suspended solid-floor polypropylene cages furnished with woodflakes. Animals were housed individually during the 24 hour exposure period and in groups of 5, by sex, for the remainder of the study.
- Diet (e.g. ad libitum): ad libitum.
- Water (e.g. ad libitum): ad libitum access to mains drinking water.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): At least 15 changes per hour.
- Photoperiod (hrs dark / hrs light): The lighting was controlled by a time switch to give twelve hours continuous light (0600 to 1800) and twelve hours darkness.

IN-LIFE DATES: From: 3 October 2012 To: 17 October 2012

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
arachis oil
Details on dermal exposure:
TEST SITE
- Preparation: On the day before treatment, the back and flanks of each animal were clipped free of hair.
- % coverage: Approximately 10 % of the total body surface area.
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated skin and surrounding hair were wiped with cotton wool moistened with arachis oil BP to remove any residual test material.
- Time after start of exposure: 24 hours.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bodyweight.

VEHICLE
- Amount(s) applied: The appropriate amount of the test material was moistened with arachis oil BP and applied as evenly as possible.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for deaths or overt signs of toxicity 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days.
After the removal of the dressings and subsequently once daily for 14 days, the test sites were examined for any evidence of primary irritation and scored according to the criteria of Draize (shown below).
Individual bodyweights were recorded prior to application of the test material on day 0 and on days 7 and 14.
- Necropsy of survivors performed: Yes; all animals were subjected to gross necropsy. Animals were killed by cervical dislocation. An external examination was performed and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded.


Draize Scale for Primary Irritation:

Erythema and eschar formation:
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

Oedema formation:
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (edges raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were observed.

DERMAL REACTIONS
Individual dermal reactions are given in Table 1 and Table 2.

Red coloured staining, which prevented evaluation of erythema, was noted at the test sites of all animals one day after patch removal.
Pale red coloured staining, which did not prevent evaluation of skin responses, was noted at the test sites of nine animals from day 2 up to day 6.
Very slight to well-defined erythema was noted at the test sites of all animals. Very slight oedema was also noted at the test sites of all females. Moderate desquamation was noted at the test sites of all males and one female. Crust formation was noted at the test sites of all females with small superficial scattered scabs also noted at the test sites of two females.
Body weight:
All animals showed the expected gains in bodyweight over the observation period.
Gross pathology:
No abnormalities were noted at necropsy.

Any other information on results incl. tables

Table 1 Individual Dermal Reactions (Male)

Animal Number

Observation

Effects Noted After Initiation of Exposure (days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

 

1-0

Erythema

Oedema

Other

?

0

0

1

0

S

1

0

D

1

0

D

1

0

D

1

0

D

0

0

D

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

1-1

Erythema

Oedema

Other

?

0

0

1

0

0

1

0

D

1

0

D

1

0

D

1

0

D

0

0

D

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

1-2

Erythema

Oedema

Other

?

0

0

1

0

S

1

0

D

1

0

D

0

0

D

1

0

D

0

0

0

0

0

D

0

0

D

0

0

D

0

0

D

0

0

0

0

0

0

0

0

0

 

1-3

Erythema

Oedema

Other

?

0

0

2

0

S

2

0

S,D

2

0

S,D

1

0

S,D

1

0

D

0

0

D

0

0

D

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

1-4

Erythema

Oedema

Other

?

0

0

1

0

S

1

0

D

1

0

D

1

0

D

1

0

D

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

? = Red coloured staining prevented the evaluation or erythema.

S = Pale red coloured staining

D = Moderate desquamation

 

Table 2 Individual Dermal Reactions (Female)

Animal Number

Observation

Effects Noted After Initiation of Exposure (days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

 

2-0

Erythema

Oedema

Other

?

0

0

2

1

S

2

1

S,C

2

1

S,C

2

1

S,C

2

1

C

1

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

0

 

2-1

Erythema

Oedema

Other

?

0

0

2

1

S

2

1

S,C

2

1

S,C

2

1

S,C

2

1

C

1

0

C

1

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

0

0

0

0

 

2-2

Erythema

Oedema

Other

?

0

0

2

1

S

2

1

C

2

1

C

2

1

C

2

1

C

1

0

C

1

0

C

0

0

C

0

0

C*

0

0

D*

0

0

D

0

0

*

0

0

*

 

2-3

Erythema

Oedema

Other

?

0

0

2

1

S

2

1

S,C

2

1

S,C

2

1

S,C

2

1

S,C

1

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

C

0

0

0

 

2-4

Erythema

Oedema

Other

?

0

0

2

1

S

2

1

C

2

1

C

2

1

C

2

1

C

1

0

C

0

0

C

0

0

C

0

0

C*

0

0

C*

0

0

C*

0

0

C*

0

0

*

? = Red coloured staining prevented the evaluation or erythema.

S = Pale red coloured staining

D = Moderate desquamation

C = Crust formation

* = Small, superficial, scattered scabs

Applicant's summary and conclusion

Interpretation of results:
other: CLP criteria not met
Conclusions:
LD50 >2000 mg/kg bodyweight
Executive summary:

The acute dermal toxicity potential of the test material to Wistar strain rats was assessed in a limit test conducted in accordance with the standardised guidelines OECD 402 and EU Method B.3 under GLP conditions.

A group of 5 male and 5 female animals was given a single dermal dose of the test material to intact skin at a dose level of 2000 mg/kg bodyweight. The test site was covered with a semi-occlusive dressing and left in place for 24 hours. The animals were subsequently observed for 14 days.

There was no mortality and no clinical signs were observed. All animals showed the expected gains in bodyweight throughout the observation period. Signs of dermal irritation noted were very slight to well-defined erythema, very slight oedema, moderate desquamation, crust formation and small, superficial, scattered scabs.

Under the conditions of this study, the acute dermal LD50 was >2000 mg/kg bodyweight and the test material requires no classification in accordance with EU criteria.