Reaction mass of 2,2'-(vinylenedi-p-phenylene)bis[5-methylbenzoxazole] and 2,2'-(vinylenedi-p- phenylene)bisbenzoxazole and 2-[4-[2-[4-(benzoxazol-2- yl)phenyl]vinyl]phenyl]-5-methylbenzoxazole

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October from 14 to 30, 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Fluorescent Brightener 219

IUPAC Name:

Fluorescent Brightener 219

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes, Chatillon sur Chalaronne (France).
- Age at study initiation: 9 weeks male and 13 - 14 weeks females.
- Housing: individually in stainless steel cages.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet, ad libitum.
- 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.
- Health check. 4 days before treatment, one flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm × 10 cm). 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.

ENVIRONMENTAL CONDITIONS
- Temperature: air-conditioned with target ranges for room temperature 17-23 °C.
- Relative humidity: 30 - 70 %
- Air changes: approximately 10 - 15 air changes per hour.
- Photoperiod: automatically controlled light cycle of 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:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Amount: 0.5 g (per animal) of test item was weighed and moistened with approximately 0.1 ml of purified water before application.
- pH check: before the study initiation date, formulation ol 1 % in water was prepared and the pH was found to be 6.

Duration of treatment / exposure:

4 hours

Observation period:

10 days

Number of animals:

1 male and 2 females

Details on study design:

TEST SITE
- Area of exposure: on the day of treatment, test item was placed on a surgical gauze patch (ca.4 cm × 4 cm). This gauze patch was applied to the intact skin of the clipped area.
- Type of wrap if used: 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: skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4 hours.

OBSERVATION TIME POINTS
- skin reaction: assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69|EEC, July 31, 1992 approximately 1, 24, 48 and 72 hours, as well as 7 and 10 days after the removal of the dressing, gauze patch and test item. To allow further examination of the test sites animal Nos. 58 and 59 were re-clipped on completion of the 48-hour examination. lf evident, corrosive or staining properties of the test item were described and recorded.
- 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.

SCORING SYSTEM
To evaluate the irritation of the test item the mean score was calculated across 3 scoring times (24,48 and72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. An animal is positive when the mean score is 2 or greater. The test is positive for irritation when at least 2 animals are positive for the same endpoint (erythema/eschar or oedema).

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 i 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:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The mean erythema/eschar score and the mean oedema score of the three animals were 0.00. No erythema and no swelling (oedema) was noted in any animal at any time.
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

COLORATION
Slight yellow staining produced by the test item of the treated skin was observed in all animals from the 1-hour reading up to 48 hours after treatment. Slight yellow staining was still evident in two animals up to the 72-hour examination and persisted in one animal up to 7 days after treatment.

Other effects:

VIABILITY/MORTALITY/CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

BODY WEIGHTS
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 individual scores

Animal no. / sex	Reaction	1 hr	24 hrs	48 hrs	72 hrs	7 days	10 days	Mean 24, 48, 72 hrs
58 M	Erythema	0	0	0	0	0	0	0.00
59 F	Erythema	0	0	0	0	0	0	0.00
60 F	Erythema	0	0	0	0	0	0	0.00
58 M	Oedema	0	0	0	0	0	0	0.00
59 F	Oedema	0	0	0	0	0	0	0.00
60 F	Oedema	0	0	0	0	0	0	0.00
58 M	Staining	*	*	*	*			-
59 F	Staining	*	*	*	*	*		-
60 F	Staining	*	*	*				-

*slight yellow staining produced by the test item

Examinations were performed at the specified times after removal of the dressing.

Note: Observations continued for 10 days due to staining present on the skin.

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC 1272/2008)

Conclusions:

Not irritating

Executive summary:

The primary skin irritation potential of the substance was investigated according to OECD guideline 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three oung 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 and 10 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 and the mean oedema score of the three animals were 0.00. The application of test item to the skin resulted in no signs of irritation.

However, yellow staining produced by the test item was present in all animals. This effect was reversible and was no longer evident 10 days after treatment, the end of the observation period for all animals. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no other clinical signs of test item related effects were observed.

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