Registration Dossier

Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
July 16 - July 30, 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): E-BW102
- Physical state: Reddish brown powder
- Storage condition of test material: Room temperature, dark place
- Stability under storage conditions: No data

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
- Source: Kitayama Labes Co., Ltd.
- Age at study initiation: 10 weeks old *.
- Weight at study initiation: 2080-2144 g *.
- Housing: Animals were individually housed in labeled aluminium cages (dimensions 31.8 x 35.8 x 49.8 cm) with stainless-steel cascade racks.
- Diet (e.g. ad libitum): Pellet diet (LRC4, Oriental Yeast Co., Ltd., lot 100420)
- Water (e.g. ad libitum): Free access to well water.
- Acclimation period: The acclimatization period was 6 days from receipt.

* Animal specifications (sex, age and body weight) are specified in the attached table.

On the day before application, the skin condition was observed, and the rabbits with healthy skin were selected.

Results of analysis for diet was provided by Oriental Yeast Co., Ltd., and the contaminants in the diet were confirmed to be within the acceptable limits established by the test facility. The water was analyzed every 6 months at Nichigo Kyushu Co., Ltd. in compliance with the water quality standard (Waterworks Law) of MHLW, Japan. The contaminants in the water were confirmed to be within the acceptable limits.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24.0 – 26.1
- Humidity (%): 50.6 - 73.2
The accommodation equipment was washed with a cage washer. The animal room was swept and wiped every day with a mop soaked in disinfectant.
- Air changes (per hr): 10 to 20 times
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: July 27, 2010 to July 30, 2010

Test system

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: The test substance was moistened with 0.3 mL of water for injection.
Controls:
other: Adjacent areas of the untreated skin of each animal served as controls.
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 0.5 grams

VEHICLE
The test substance was moistened with 0.3 mL of water for injection.
Duration of treatment / exposure:
Single application/ 4 hours
Observation period:
72 hours
Number of animals:
3 males.
Details on study design:
TEST SITE
On the day before application (approximately 24 hours before treatment, the skin on the back was shaved with electric clippers. Before application on the day of treatment, an Elizabeth color was installed on the rabbits. On the back of the rabbits an area of 2.5x2.5 cm was established.

The test substance was placed on a patch of 2.5x2.5 cm in size and placed on the application area. The patch was fixed with a non-woven adhesive bandage of about 5x5 cm. The application site was occluded by the adhesive elastic bandage.

REMOVAL OF TEST SUBSTANCE
Four hours after the application, the occlusion was removed and the application site was cleaned with absorbent cotton moisturized with lukewarm water. Then, the Elizabeth collar was removed.

The animals were euthanized by an intravenous overdose of pentobarbital sodium after the final observation.

OBSERVATIONS
- Clinical observation: Observation was performed twice, before and after application and once daily on the other days.
- Body Weight: Body weight was measured before application and 72 hours after patch removal (final observation day).
- Irritation:
The skin reactions were assessed at approximately 1, 24, 48 and 72 hours after patch removal. The irritation scores and a description of all other (local) effects were recorded.

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

Erythema and eschar formation:
0: No erythema
1: Very slight erythema (barely perceptible)
2: Well-defined erythema
3: Moderate to severe erythema
4: Severe erythema (beef redness) *
*. Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) was given.

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

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
mean
Remarks:
animal #1, #2 and #3
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Remarks:
animal #1, #2 and #3
Time point:
other: 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Remarks:
animal #1, #2 and #3
Time point:
other: 24, 48, and 72 hours
Score:
0
Max. score:
8
Irritant / corrosive response data:
No skin reaction was observed in any animal at the application site at any observation time.

Tables containing individual and mean irritation scores are specified in the attached table.
Other effects:
No clinical signs were observed in any animal throughout the observation period.

No abnormal body weight gain was observed in any animal throughout the observation period.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
A skin irritation/corrosion study with E-BW102 in the rabbit (4-hour semi-occlusive application, 3 males/dose) was conducted according to OECD 404 and GLP guidelines. Based on the results of this study E-BW102 does not have to be classified and has no obligatory labelling requirement for skin irritation according to Regulation (EC) No. 1272/2008.