Reaction mass of copper complex of [(2,6-difluoroheterocycl-4-yl)amino]hydroxy{[2-hydroxy-3-sulfonato-5-(vinylsulfonyl)phenyl]diazenyl}naphthalene sulfonic acid, dialkali salt and copper complex of [(2,6-difluoroheterocycl-4-yl)amino]-hydroxy{[2-hydroxy-3-sulfonato-5-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl]diazenyl}naphthalene sulfonic acid, trialkali salt

Endpoint summary

Description of key information

A study was conducted to assess the skin irritation potential of the test substance to the rabbit skin according to EU Method B.4, EPA OPPTs 870.2500 and OECD Guideline 404 in compliance with GLP. An eye irritation study in rabbits was performed according OECD 405 and EU Method B.5, following GLP guidelines. The test compound caused irreversible discolorations in the eye of the animal. This is not considered an adverse effect but substance specific staining.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-10-21 to 2002-11-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Conducted in accordance with EU Method B.4, OECD 404 and EPA OPPTs 870.2500, in accordance with GLP.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, 88353 Kiβlegg
- Weight at study initiation: 2.5 - 2.9 kg
- Diet: ssniff K-H (V2333), ad libitum and hay (approx. 15 g daily)
- Water: water from automatic water dispensers, ad libitum
- Acclimation period: 1 wk

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3 °C except for short lasting deviations due to disturbance of air condition
- Humidity (%): 50±20 °C except for short lasting deviations due to disturbances of air condition
- Photoperiod (hrs dark / hrs light): 12 h dark/12 h light

IN-LIFE DATES: From: Oct 29 2002 To: Nov 08 2002

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 h

Observation period:

24, 48 and 72 h after removal of the patches

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: Dorsal region of the body
- % coverage: 25 cm2
- Type of wrap if used: Yes, The test substance was evenly distributed over the whole surface of a 2.5 x 2.5 cm cellulose patch on a piece of surgical plaster. The plaster was fixed to the prepared skin area and then covered with a semi-occlusive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, with warm water
- Time after start of exposure: 4 h

SCORING SYSTEM: Erythema, eschar formation and edema were evaluated numerically according to the score of DRAIZE

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24, 48 and 72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24, 48 and 72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

Reversibility of any observed effect: Changes fully reversible within 1 days

Other effects:

No signs of systemic toxicity.

1 animal out of 3 showed a very mild erythrema 30-60 minutes after patch removal, which was fully reversibly until day 1 observation (24 hours after patch removal).

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, the test substance was considered to be non-irritating to rabbit skin

Executive summary:

A study was conducted to assess the skin irritation potential of test substance to the rabbit skin according to EU Method B.4, EPA OPPTs 870.2500 and OECD Guideline 404 in compliance with GLP.

Three animals were treated with 0.5 g of test substance pasted in deionised water using semi-occlusive patches for 4 h. After the exposure period all remnants of the test substance were carefully removed from the skin with warm tap water. The skin was evaluated for erythema, eschar formation and edema numerically according to the score of Draize system for up to 72 h after patch removal.

Very slight signs of erythema was observed in one animal 30 to 60 mintes after removal of the patch which was reversible.

Under the test conditions, the test substance was considered to be non-irritating to rabbit skin.

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:

13 November 2002 to 06 January 2003

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Conducted in accordance with OECD 405, EU Method B.5 and EPA OPPTs 870.2400 and GLP; only one animal tested.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH, 88353 Kiβlegg
- Weight at study initiation: 3.4 kg
- Diet: ssniff K-H (V2333), ad libitum and hay (approx. 15 g daily)
- Water: From automatic water dispensers, ad libitum
- Acclimation period: 1 wk

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±3 °C (except short lasting deviations doe to disturbances of air condition)
- Humidity (%): 50±20 °C (except short lasting deviations doe to disturbances of air condition)
- Photoperiod (hrs dark / hrs light): 12 h dark/12 h light

IN-LIFE DATES: From: Nov. 19, 2002 To: Dec. 10, 2002

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

1, 24, 48 and 72 hours after administration of drug. The eye was also further examined after 7, 14 and 21 days due to effects being observed after 72 hours.

Number of animals or in vitro replicates:

1

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, 24 h after administration the treated eyes were washed out thoroughly with isotonic saline at approx. 37 °C
- Time after start of exposure: 24 h

SCORING SYSTEM: Lesions in cornea, iris or conjunctivae were graded numerically "Scale for scoring ocular reactions"

TOOL USED TO ASSESS SCORE: 0.01 % fluorescein-sodium solution

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24, 48 and 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48h

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 1 h

Score:

0

Max. score:

2

Remarks on result:

other: all other time points were not assessable, due to dark violet discoloration caused by the test substance

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24, 48, and 72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24, 48 and 72 h

Score:

0

Max. score:

3

Remarks on result:

other: not assessable, due to dark violet discoloration causes by the test substance

Irritant / corrosive response data:

From one hour up to 2 days after application the animal showed slight up to severe swellings with more than half closed lids. The cornea of the animal showed diffuse areas of opacity 1 day after application.
The irritations were attended by dark violet discolored eye discharge 1 hour after administration.
Additionally, conjunctiva, iris, nictitating membrane and sclera were discolored violet or dark violet from day two up to the end of the study.
Due to the violet or dark violet discoloration of the conjunctiva and iris the animal could not be assessed for redness of conjunctiva and iris.

Other effects:

Discoloration of conjunctiva, iris, nictitating membrane and sclera not fully reversible within 21 days.

From one hour up to 2 days after application the animal showed slight up to sever swellings with more than half closed lids. The irritations were attended by dark violet discoloured eye discharge 1 hour after administration. Additionally conjunctiva, iris, nictitating membrane and sclera were discoloured violet or dark violet from day two up to the end of the study and therefore the conjunctiva and iris could not be assessed for redness.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The applicant concluded that the persistent discolouration of the eye is a dye specific effect and does not require classification.

Executive summary:

A study was conducted to assess the eye irritation potential of test substance to the rabbit eye according to EU Method B.5., US EPA OPPTs 870.2400 and OECD Guideline 405 in compliance with GLP.

One animal received 0.1 g of undiluted test substance into the conjunctival sac of the left eye. The other eyes served as untreated control. 24 h after the administration the eyes were washed thoroughly with isotonic saline at 37 °C. The eyes were also washed out at designated examination times at which discharge was observed or a corneal examination with fluorescein was performed.

The eyes were examined 1, 24, 48 and 72 h after administration of the test substance and due to observation of continued effects at 72 hours, the eyes were also examined after 7, 14 and 21 days. At 24h, 72 h, 7 days, 14 days and 21 days, the eyes were further examined for corneal lesions under UV light after instillation of one drop of a 0.01 % fluorescein-sodium solution. Lesions in cornea, iris or conjunctivae were graded numerically (Scale for scoring ocular reactions). All other changes or toxic effects were recorded when apparent.

From one hour up to 2 days after application the animal slight up to severe swellings with more than half closed lids. The cornea showed diffuse areas of opacity after application. The irritations were attended by a dark violet discoloured eye discharge and the conjunctiva, iris, nictitating membrane and sclera were discoloured violet or dark violet from day 2 up to the end of the study. Only scores for the opacity of the cornea and chemosis of the conjunctiva were able to be calculated. These were means from 24, 48 and 72 hours of 0.33 and 1.33 respectively. Due to the continued violet discoloration of the eye, the animals could not be assessed for redness of the conjunctiva and iris.

Under the test conditions, the test substance showed potential risk of serious damage to eyes according to the classification criteria of Directive 2001/59/EC. However, the irreversible discoloration of the conjunctiva is not considered an adverse effect according to Regulation (EC) 1272/2008, as this discoloration does not lead to an impaired sight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

A study was conducted to assess the skin irritation potential of test substance to the rabbit skin according to EU Method B.4, EPA OPPTs 870.2500 and OECD Guideline 404 in compliance with GLP.

Three animals were treated with 0.5 g of the test substance pasted in deionised water using semi-occlusive patches for 4 h. The skin was washed and evaluated for erythema, eschar formation and edema numerically according to the score of Draize system for up to 72 h after patch removal.

Very slight signs of erythema was observed in one animal 30 to 60 minutes after removal of the patch which was reversible. However, under the test conditions, the test substance was considered to be non-irritating to rabbit skin.

Eye irritation

A study was conducted to assess the eye irritation potential of test substance to the rabbit eye according to EU Method B.5., US EPA OPPTs 870.2400 and OECD Guideline 405 in compliance with GLP.

1 animal received 0.1 g of an undiluted test substance into conjunctival sac of the left eye. 24 h after the administration the eyes were washed. Eyes were examined 1, 24, 48 and 72 h after administration of the test substance and due to observation of continued effects at 72 hours, the eyes were also examined after 7, 14 and 21 days. At 24h, 72 h, 7 days, 14 days and 21 days, the eyes were further examined for corneal lesions under UV light after instillation of one drop of a 0.01 % fluorescein-sodium solution. Lesions in cornea, iris or conjunctivae were graded numerically and all other changes or toxic effects were recorded when apparent.

From one hour up to 2 days after application the animal slight up to severe swellings with more than half closed lids. the cornea showed diffuse areas of opacity after application. The irritations were attended by a dark violet discoloured eye discharge and the conjunctiva, iris, nictitating membrane and sclera were discoloured violet or dark violet from day 2 up to the end of the study. Only scores for the opacity of the cornea and chemosis of the conjunctiva were able to be calculated. These were means from 24, 48 and 72 hours of 0.33 and 1.33 respectively. Due to the continued violet discoloration of the eye, the animals could not be assessed for redness of the conjunctiva and iris.

Under the test conditions, the test substance showed potential risk of serious damage to eyes according to the classification criteria of Directive 2001/59/EC. However, the irreversible discoloration of the conjuctiva is not considered an adverse effect according to Regulation 1272/2008, as this discoloration does not lead to an impaired sight.

Justification for selection of skin irritation / corrosion endpoint:
GLP guideline study

Justification for selection of eye irritation endpoint:
GLP guideline study

Justification for classification or non-classification

The available data suggest that test substance is not classified as an irritant to the skin or eyes.