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

Eye irritation

Administrative data

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.

Cross-reference

Reason / purpose for cross-reference:

reference to same 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 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in the study report): Reaktiv Rot F00-0124

Test animals / tissue source

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

Test system

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

Results and discussion

In vivo

Results

Results 1

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

Results 2

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

Results 3

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

Results 4

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.

Any other information on results incl. tables

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.

Applicant's summary and conclusion

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.