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

Endpoint summary

Description of key information

Not skin/eye irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted April 24, 2002

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

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.

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

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

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.

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.

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).

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March from 04 to 24, 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted April 24, 2002

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

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: 11 weeks male and 12 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.

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:

14 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

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible within: 72 hours

Remarks on result:

no indication of irritation

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 of the three animals was 0.67, 0.67 and 0.00, respectively and the mean oedema score was 0.00, 0.00 and 0.00, respectively.
Very slight erythema was observed in two animals at the 1-hour reading. Very slight erytnema was observed in two animals 24 and 48 hours after treatment.
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

COLORATION
Green staining produced by the test item of the treated skin was observed in all animals from 1 to 48 hours after treatment and persisted in one animal up to 72 hours and in another animal up to the end of the observation time.

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.

Skin irritation individual scores

Animal N / sex	Reaction	1 hr	24 hrs	48hrs	72 hrs	7 days	10 days	14 days	Mean 24, 48, 72 hrs
67 M	Erythema	1	1	1	0	0	0	0	0.67
68 F	Erythema	0	1	1	0	0	0	0	0.67
69 F	Erythema	1	0	0	0	0	0	0	0.00
67 M	Oedema	0	0	0	0	0	0	0	0.00
68 F	Oedema	0	0	0	0	0	0	0	0.00
69 F	Oedema	0	0	0	0	0	0	0	0.00
67 M	Staining	*	*	*	-	-	-	-	-
68 F	Staining	*	*	*	*	-	-	-	-
69 F	Staining	*	*	*	*	*	*	*	-

*light green staining produced by the test item

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

Note: Observations continued for 14 days due to staining present on the skin.

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, 10 and 14 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, 0.67 and 0.00, respectively and the mean oedema score was 0.00, 0.00 and 0.00, respectively.

The application of test item to the skin resulted in mild signs of irritation. These included erythema. These effects were reversible and were no longer evident 72 hours after treatment. The test item caused green staining of the treated ski in one animal throughout the observation time. No corrosive effects were ed skin of any animal at any of the measuring intervals and no other clinical signs of substance 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).

Reference 3

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 30 to April 08, 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted April 24, 2002

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

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: 11 weeks.
- 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.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Amount: 0.5 g of test item was moistened with approximately 0.1 ml of purified water before application.
- Application: the test item was prepared in a dark room, then covered with an aluminium foil before treatment. lt was applied in the dark animal room (main light out).
- pH check: before the study initiation date, formulation ol 1 % in water was prepared and the pH was found to be 5.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

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 and 72 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

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Reversibility:

fully reversible within: 72 hours

Remarks on result:

no indication of irritation

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 was 0.00 for all three animals. No erythema and no swelling (oedema) was observed 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 at the 1-hour reading and persisted in one animal up to the 24-hour examination.

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.

Skin irritation individual scores

Animal no. / sex	Reaction	1 hr	24 hrs	48 hrs	72 hrs	Mean 24, 48, 72 hrs
10 M	Erythema	0	0	0	0	0.00
11 F	Erythema	0	0	0	0	0.00
12 F	Erythema	0	0	0	0	0.00
10 M	Oedema	0	0	0	0	0.00
11 F	Oedema	0	0	0	0	0.00
12 F	Oedema	0	0	0	0	0.00
10 M	Staining	*	*
11 F	Staining	*
12 F	Staining	*

* slight yellow staining produced by the test item

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 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 was 0.00 for all three animals.

The application of test item to the skin resulted in no signs of irritation. However slight yellow staining of the treated skin was observed in all animals at the 1 -hours reading and persisted in one animal up to the 24 -hour examination. No corrosive effects were observed on skin of any animal at any of the measuring intervals and no other clinical signs of substance 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).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January from 12 to 15, 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

other: FDA guideline, federal register 38, No. 187, 27.9.1973, p 27019

GLP compliance:

no

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 15 - 19 kg
- Housing: single cage.
- Diet. standard diet (ERKA 8300), ad libitum.
- Water: ad libitum.

Vehicle:

other: polyethyene glycol 400

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg of the substance powder-form (mixed with 0.25 ml of polyethyene glycol 400)

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

6 rabbits

Details on study design:

APPLICATION
- Treatment: test compound was instilled into the conjunctival sac of the left eye. The eyelids were then held closed for 1 second.

REMOVAL OF TEST SUBSTANCE
- Washing: physiological saline.
- Time after start of exposure: 24 hours.

TOOL USED TO ASSESS SCORE: magnifying glass and at 48 and 72 hours after additional installation of a drop of flurescein sodium in the dilution 1: 10000, i.e. 0.01%

SCORING SYSTEM
According to the FDA guideline, federal register 38, No. 187, 27.9.1973, p 27019.
The assessment was perfromed after 1, 7, 24, 48 and 72 hours after application.

Cornea
A Opacity-degree of density (area most dense taken for reading)
No opacity 0
Scattered or diffuse area, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris invisible 4

B Area of cornea involved
One quarter (or less) but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4

A × B × 5 Total maximum = 80

Iris
A Values
Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, hemorrhage, gross destruction (any or all of these) 2

A × 5 Total maximum = 10

Conjunctivae

A Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

B Chemosis
No swelling 0
Any swelling above normal (includes nictitating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids about half closed to completely closed 4

C Discharge
No discharge 0
Any amount different From normal (does not include small amounts observed in inner cunthas of normal animals) 1
Discharge with moistening oi the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, and considerable area around the eye 3

(A + B + C) × 2 Total maximum = 20

To determine the irritation index, the scores of the cornea (a), the iris (b) and the conjunctiva (c) were added for each rabbit at the respective reading time. From the values of all rabbits the mean value was formed at the respective reading time. The highest index determined at one of the readings was used for the classification according to the following verbal evaluation classes:
0 - 10 non irritant
11 - 25 slightly irritant
26 - 56 moderately irritant
57 - 110 severely irritant

Irritation parameter:

cornea opacity score

Basis:

animal:

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Remarks:

score reasonably expected < 1

Irritation parameter:

iris score

Basis:

animal:

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Remarks:

score reasonably expected < 1

Irritation parameter:

conjunctivae score

Basis:

animal:

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Remarks:

score reasonably expected < 2

Irritation parameter:

chemosis score

Basis:

animal:

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Remarks:

score reasonably expected < 2

Irritant / corrosive response data:

The highest irritation index was 10 after 1 hour.

Interpretation of results:

other: non irritant

Conclusions:

Non irritant

Executive summary:

The test substance eye irritation potential was assayed in 6 rabbits. The test was carried out in accordance with FDA guideline, federal register 38, No. 187, 27.9.1973, p 27019. 100 mg of the substance powder-form (mixed with 0.25 ml of polyethyene glycol 400) were instilled into the conjunctival sac of the eye. After 24 hours, eyes were rinsed with physiological saline. The rabbits were examined at 1, 7, 24, 48 and 72 hours after application of the test compound.

The highest irritation index resulted to be 10 after 1 hour, thus substance can be considered as not eye irritating.

Conclusion

Non irritant

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

SKIN IRRITATION

The skin irritation potential was investigated in several experiments, according to OECD guideline 404.

In all cases, only slight irritating effects were seen, which were recorded after observation period. Yellow/green staining of the treated skin was observed. 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.

The additional supporting study reports comparable results; the test was carried out according to the FDA guideline 1500.41, federal register 38, No. 187, 27.9.1973, p 27019. Two test sites of the back were used on each rabbit; before the application of the test compound, one hand site was abraded. Primary irritation index resulted to be 0.8, thus the substance was considered as slighlty irritating.

EYE IRRITATION

Only one experiment conducted with Fluorescent Brightener 219 is available on the eye irritation potential. The experiment was conducted in 6 rabbits, in accordance with FDA guideline, federal register 38, No. 187, 27.9.1973, p 27019. The highest irritation index resulted to be 10 after 1 hour, thus substance was judged to be not eye irritating.

In order to support the study outcomes, the available data on the one of the substance's constituents (i.e. constituent 02) have also been taken into conderation.

The study was performed with the New Zealand White rabbit, in accordance with OECD guideline 405 and EU Method B.5. A single application of 0.1 ml (67.3 mg) of the test material was administered to the non irrigated eye of three rabbits. The Maximum Group Mean Score was 4.1 and the related classification was as mild irritant (class 4 on a 1 -8 SCALE).

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.2 Skin corrosion/irritation section, skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

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

According to the CLP Regulation (EC 1272/2008), serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance, which is not fully reversible within 21 days of application.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all of the three tested animals. All the reactions resulted to be fully reversible withing 72 hours. Therefore, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC 1272/2008).