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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental starting date: 27 February 2008 and Experimental completion date: 06 March 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
- Animals: Young Adult New Zealand White Rabbit, SPF
- Rationale: Recognized by international guidelines as a recommended test system.
- Number of Animals per Test: 3 (Animals of both sexes were used)
- Age when treated: 17 weeks (male) and 13 weeks (females)
- Identification: By unique cage number and corresponding ear number.
- Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
- Allocation: Male No. 75 and Female Nos. 76, 77

- Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 17-23 °C and for relative humidity between 30-70 % (values above 70 % during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.
- Accommodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 36/07, Provimi Kliba AG) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 70/07) provided by Provimi Kliba AG, CH-4303
Kaiseraugst.
- Water: Community tap water from Füllinsdorf ad libitum.

Test system

Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 gram
Duration of treatment / exposure:
4 hours
Observation period:
72 hours
Number of animals:
3 animals of both sexes were used.
Details on study design:
Four days before treatment, the left flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm x 10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was clipped again.

Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.
On the day of treatment, 0.5 g of TKK 30302 (XU 3560 / LMB 6452) was placed on a surgical gauze patch (ca. 2.5 cm x 2.5 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around
the abdomen and anchored with tape.

The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site.

SCORING SYSTEM:
COMMISSION DIRECTIVE 2004/73/EC, APRIL 29, 2004
Grading of Skin Reactions

ERYTHEMA AND ESCHAR FORMATION
- No erythema 0
- Very slight erythema 1
- Well-defined erythema 2
- Moderate to severe erythema 3
- Severe erythema (beef redness) or eschar
- formation (injuries in depth preventing erythema) reading 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 mm) 3
- Severe oedema (raised more than 1 mm and extending beyond the area of exposure) 4

Results and discussion

In vivo

Results
Irritation parameter:
overall irritation score
Basis:
mean
Remarks:
3 rabbits (1 male and 2 females)
Score:
0
Max. score:
0
Remarks on result:
other: No erythema and no edema were observed.
Irritant / corrosive response data:
Irritation
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately.
The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

Corrosion
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.
Other effects:
- Viability / Mortality / Clinical Signs: No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
- Coloration: No staining produced by the test item of the treated skin was observed.
- Body weight: The body weights of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

Body weights in grams.

 Animal N° Sex  First day of acclimatization  Day of treatment  Last day of observation 
 75  male  2926  3039 3065 
 76  female  2430  2746 3046 
 77  female  2474  2600 2774 

Skin irritationscores-Individual values.

 Animal number Sex  Evaluation Interval*  Erythema  Edema  Scaling 
75   M   1 hour  0  0  0
 76  F   1 hour  0  0
 77  F   1 hour  0  0
 75  M   24 hours  0  0  0
 76  F   24 hours  0  0
 77  F   24 hours  0  0  0
 75  M  48 hours  0  0  0
 76  F  48 hours  0  0  0
 77  F  48 hours  0  0  0
 75  M  72 hours  0  0  0
 76  F  72 hours  0  0  0
 77  F  72 hours  0  0  0

*Examination were performed at the specified times after ramoval of the dressing.

SKIN IRRITATION SCORES – INDIVIDUAL MEAN VALUES AFTER 24, 48 AND 72 HOURS

 Animal number Sex  Erythema  Oedema 
 75  M  0.00  3  0.00  3
 76  F  0.00  3  0.00  3
 77  F  0.00  3  0.00  3

N= number of available data points.

SKIN IRRITATION SCORES – ASSESSMENT ACCORDING TO EC GUIDELINES

 Evaluated intervals  Erythema Oedema 
 24 hours   Not irritating   Not irritating
 48 hours   Not irritating   Not irritating
 72 hours   Not irritating   Not irritating

Individual Findings (for animal 75, 76 and 77).

After 1 hour:

Erythema: NO ABNORMAL FINDINGS NOTED

Oedema: NO ABNORMAL FINDINGS NOTED

Flaking: NO ABNORMAL FINDINGS NOTED

Staining: NO STAINING PRESENT

After 24 hours:

Erythema: NO ABNORMAL FINDINGS NOTED

Oedema: NO ABNORMAL FINDINGS NOTED

Flaking: NO ABNORMAL FINDINGS NOTED

Staining: NO STAINING PRESENT

After 48 hours:

Erythema: NO ABNORMAL FINDINGS NOTED

Oedema: NO ABNORMAL FINDINGS NOTED

Flaking: NO ABNORMAL FINDINGS NOTED

Staining: NO STAINING PRESENT

After 72 hours:

Erythema: NO ABNORMAL FINDINGS NOTED

Oedema: NO ABNORMAL FINDINGS NOTED

Flaking: NO ABNORMAL FINDINGS NOTED

Staining: NO STAINING PRESENT

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), the test item is considered to be “not irritating” to rabbit skin.
Executive summary:

The primary skin irritation potential of the test item was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours after removal of the dressing.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately.

The test item did not elicit any skin reactions at the application site of any animal at any of the observation times (all scores 0). The individual mean score for erythema/eschar and oedema for each of the three animals was therefore 0.

The test item caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed.

Thus, the test item did not induce significant or irreversible damage to the skin.

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), the test item is considered to be “not irritating” to rabbit skin.