Leucophor 1111X

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From August 24th to September 06th, 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted according to internationally accepted testing guidelines and performed in compliance with Good Laboratory Practice. Justification for read across approach is given in the endpoint summary and in the read across justification report attached to the Section 13 of this dossier.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted April 24, 2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

April 29, 2004

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH Stolzenseeweg 32-36 D-88353 Kisslegg / Germany.
- Age at study initiation: 11-12 weeks (male); 10-11 weeks (females).
- Weight at study initiation: 1687 - 2454 g
- Housing: individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4642 (batch no. 35/05, Provimi Kliba AG) were provided for gnawing.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 34/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water: community tap water from Füllinsdorf, ad libitum.
- Acclimation period: under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature: ranges for room temperature 17-23 °C.
- Humidity: 30-70 %
- Air changes: approximately 10-15 air changes per hour.
- Photoperiod: animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark.
- Other: music was played during the daytime light period.
Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at testing laboratory.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST ITEM PREPARATION
0.5 g (per animal) of the test item was weighed as delivered by the Sponsor and then moistened with approximately 0.5 ml of purified water before application.
The pH of the test item was measured before the study initiation date. A formulation of a 1 % (w/w) solution was prepared. The pH was found to be 8.07.

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 (animals of both sexes were used)

Details on study design:

As it was suspected that the test item might produce irritancy, a single animal (one female) was treated first. As neither a corrosive effect nor a severe irritant effect was observed after the 4-hour exposure, the test was completed using the two remaining animals for an exposure period of four hours.

TEST SITE
- Area of exposure: 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.
- Type of wrap if used: on the day of treatment, 0.5 g of test item was placed on a surgical gauze patch (ca. 4 cm x 4 cm). The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing: the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site. The test item could not be removed completely, so that remnants were still present at the 1-hour reading in all animals. However, any reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4 hours.

OBSERVATIONS
- Viability/Mortality: daily from acclimatization of the animals to the termination of test.
- 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.

NECROPSY
No necropsy was performed on the animals sacrificed at termination of observation.
All rabbits were sacrificed by an intravenous injection of Vetanarcol into the ear vein at a dose of at least 1 ml/kg body weight (equivalent to 162 mg sodium pentobarbitone/kg body weight) and discarded.

SCORING SYSTEM
The skin reaction was assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004, approximately 1, 24, 48 and 72 hours, as well as 7 days after the removal of the dressing, gauze patch and test item.
If evident, corrosive or staining properties of the test item were described and recorded.

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 (beet 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

Results 1

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 2.3

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 2.3

Irritant / corrosive response data:

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 mean erythema/eschar score of the three animals was 0.67, 1.00 and 1.33, respectively and the mean oedema score was 0.00 for all three animals, respectively.
Well-defined to very slight erythema was noted in all animals from the 1- to the 48-hour reading and very slight erythema persisted in two animals up to the 72-hour observation.
Very slight swelling was observed in two animals at the 1-hour reading.
Scaling was present in one animal at the 72-hour reading.
No abnormal findings were observed on the treated skin of any animal 7 days after treatment, the end of the observation time.

COLORATION
No staining produced by the test item of the treated skin was observed.

CORROSION
Neither alterations of the treated skin were 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.
The body weights of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

SKIN IRRITATION SCORES

Animal N.	Sex	Reaction	Evaluation interval					Mean 24, 48 and 72 hrs
			1 hr	24 hrs	48 hrs	72 hrs	7 days
19	M	Erythema	2	1	1	0	0	0.67
20	F	Erythema	2	1	1	1	0	1.00
21	F	Erythema	2	2	1	1	0	1.33
19	M	Oedema	1	0	0	0	0	0.00
20	F	Oedema	0	0	0	0	0	0.00
21	F	Oedema	1	0	0	0	0	0.00
19	M	Scaling	0	0	0	0	0
20	F	Scaling	0	0	0	0	0
21	F	Scaling	0	0	0	X	0

x = scaling present

INDIVIDUAL FINDINGS

	ANIMAL N. 19, MALE	ANIMAL N. 20, FEMALE	ANIMAL N. 21, FEMALE
After 1 hour	Erythema: well-defined erythema Oedema: very slight swelling Flaking: no abnormal findings noted Staining: no abnormal findings noted Other: remnants of the test item present	Erythema: well-defined erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted Other: remnants of the test item present	Erythema: well-defined erythema Oedema: very slight swelling Flaking: no abnormal findings noted Staining: no abnormal findings noted Other: remnants of the test item present
After 24 hours	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: well-defined erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted
After 48 hours	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted
After 72 hours	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: scaling present Staining: no abnormal findings noted
After 7 days	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

not irritating

Executive summary:

The primary skin irritation potential of the substance 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, as well as 7 days 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 mean erythema/eschar score of the three animals was 0.67, 1.00 and 1.33, respectively and the mean oedema score was 0.00 for all three animals, respectively.

The application of test item to the skin resulted in mild signs of irritation. These included erythema, oedema and scaling. These effects were reversible and were no longer evident 7 days after treatment, the end of the observation period for all animals. The test item caused no staining of the treated skin, but could not be removed completely by washing with lukewarm water at the removal of the dressing, so that remnants were still present at the 1-hour reading in all animals. However, any reactions (erythema) were clearly visible at that time. 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.

Conclusion

Mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions.

In conclusion, the test item does not meet the criteria ti be classified as irritating, according to the CLP (EC 1272/2008) Regulation.