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Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 25, 2007 - May 09, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study conform with Directive 2004/73/EC, B.4, OECD Guideline No. 404

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report date:
2007

Materials and methods

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

Test material

Constituent 1
Details on test material:
Test system: Young Adult New Zealand White Rabbit, SPF
Rationale: Recongnized by the international guidelines as the recommended test system.
Source: Harlan Netherlands BV, Kreuzelweg 53, NL-5961 NM Horst / The Netherlands Postbus 6174, NL-5960 AD
Horst / The Netherlands
See also information below "Details on test animals"

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Test system: Young Adult New Zealand White Rabbit, SPF
- Number of animals per test: 3 (Animals of both sexes were used)
- Age at treatment: 13 weeks (male), 12, 14 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. 61, Female Nos. 62, 63
- Weight at study initiation: No data
- Housing: Individual in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4642 were provided for gnawing
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum.
- Water (e.g. ad libitum): Community tap water

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23°C (Air-conditioned)
- Humidity (%): 30-70% , 10-15 air changes per hour
- Photoperiod: 12 hours light and 12 hours dark
Music was played during the daytime light period.

Test system

Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
water
Controls:
no
Amount / concentration applied:
0.5 g/ animal
Duration of treatment / exposure:
1-and 4 hours
Observation period:
1, 24, 48 and 72 hours, as well as 7 days after exposure.
Number of animals:
3 (Animals of both sexes were used)
Details on study design:
TREATMENT:
Four days before treatment, the left and /or right 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 prior to test item administration and re-grown fur was clipped on the selected application site.
On the day of treatment, 0.5 g of AES 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 1- and 4 hours, respectively. Then the dressing was removed and the skin was flushed with lukewarm tap water to
clean the application site so that any reactions (erythema) were clearly visible at that time.
OBSERVATIONS.
Viability/Mortality and Clinical signs: Daily from acclimatization of the animals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation.
SKIN OBSERVATION TIME POINTS AND TERMINATION:
The skin reaction was assessed at approximately 1, 24 , 48 and 72 hours, as well as 7 days after exposure (removal of the dressing, gauze patch and test item).
NECROPSY:
All rabbits were sacrificed by an intravenous injection of Pentobarbitone into the ear vein at a dose of at least 1 mL/kg body weight (equivalent to 16mg sodium pentobarbitone/kg body weight) and discarded.



Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 4- hours exposure
Score:
0 - 0.33
Max. score:
0.67
Reversibility:
fully reversible
Remarks on result:
other: no longer evident 72 hours or 7 days after treatment.
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 4-hour exposure
Score:
0
Reversibility:
fully reversible
Irritant / corrosive response data:
No abnormal findings were observed on the treated skin of any animal 7 days after treatment, the end of the observation time.
Neither alterations of the test item of the treated skin was observed nor were corrosive effects evident on the skin.
Other effects:
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
No staining produced by the test item of the treated skin was observed.
The body weights of all rabbits were considered to be within the normal range of variability.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Base upon the referred classification criteria (commission Directive 2001/59/EC of August 2001), AES is considered to be "Not Irritation" to rabbit
skin.
Executive summary:

The mean score was calculated across 3 scoring time for each animal for erythema/eschar grades and for oedema grades, seperately. The erythema / eschar score after 1- hour exposure in one animal was 1.00 and the oedema score was 0.00. The mean erythema/eschar score after a 4- hours exposure in the three animals was 0.33, 0.67 and 0.00, respectively and mean oedema score was 0.00 for each of the three animals.