pentasodium 4-hydroxy-7-[(sulfonatomethyl)amino]-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]-8-[(2-sulfonato-4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2-sulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From Jul. 02, 2002 to Aug. 29, 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study was conducted according to EU Method B.5. OECD Guideline 405 and OPPTS 870.2400 in compliance with GLP. This study was carried out on one rabbit only, as the classification according to Directive 67/548/EEC states that the irreversible discoloration of the eye leads to a classification as R41. As the number of animals is not sufficient according to the guidelines if the substance does not cause damage to eyes, another study for eye irritation was added as read-across for this endpoint. This substance is structurally very similar to the registered substance and showed the same effect on eyes. The classification according to Council Directive 67/548/EEC and according to Regulation (EC) No 1272/2008 differs due to changes in the criteria for eye irritation in the new CLP regulation. The substance was classifies as "Xi Risk phrases: R41" due to persistent (21 days) discoloration of the nictitating membrane of one rabbit. In the new CLP regulation, the criteria to serious eye damage was reversed to "discoloration of the cornea", hence, this dye substance has no longer to be classified to be hazardeous to eyes according to REGULATION (EC) No 1272/2008.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): Reaktiv-Scharlach F01-0467

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: 3.79 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): 20±3 °C
- Humidity (%): 50±20 °C
- Photoperiod (hrs dark / hrs light): 12 h dark/12 h light

IN-LIFE DATES: From: Aug. 06, 2002 To: Aug. 27, 2002

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg

Duration of treatment / exposure:

Single treatment

Observation period (in vivo):

72 h. Since effects were still present in the treated eye after 72 h, further examinations were carried out after 7, 14 and 21 d.

Number of animals or in vitro replicates:

1 rabbit

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

Remarks:

(116)

Time point:

other: 24, 48, 72 h (mean)

Score:

0

Max. score:

4

Reversibility:

other: no corneal score observed

Remarks on result:

other: -

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

(116)

Time point:

other: 24, 48, 72 h (mean)

Score:

0

Max. score:

2

Reversibility:

other: no iris score observed

Remarks on result:

other: -

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

(116)

Time point:

other: 24, 48, 72 h (mean)

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: (after 4 d)

Remarks on result:

other: -

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

(116)

Time point:

other: 24, 48, 72 h (mean)

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: (after 7 d)

Remarks on result:

other: -

Results 5

Irritation parameter:

other: Discharge

Basis:

animal #1

Remarks:

(116)

Time point:

other: 1 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: (after 1 d)

Remarks on result:

other: -

Irritant / corrosive response data:

- 1 h after administration the conjunctiva of the animal showed slight swelling (including nictitating membrane) and some blood vessels were definitely hyperaemic (injected). Additionally, serous discharge discolored by the test compound with moistening of the lids and hairs just adjacent to the lids was noted.
- 24 h up to 48 h after administration the conjunctiva of the animal still showed some blood vessels which were definitely hyperaemic (injected).
- 72 h after administration a slight swelling of the conjunctiva was observed again. 24 h up to 21d after administration the nictitating membrane was discoloured by the test compound. This discoloration was not reversible, the study was terminated at Day 21 after administration.

Other effects:

None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Council Directive 67/548/EEC: Under the test conditions, the test substance showed potential risk of serious damage to the rabbit eye due to the persistent discoloration of the eye.

Regulation (EC) No 1272/2008: not irritating

The classification according to Council Directive 67/548/EEC and according to Regulation (EC) No 1272/2008 differs due to changes in the criteria for eye irritation in the new CLP regulation. The substance was classifies as "Xi Risk phrases: R41" due to persistent (21 days) discoloration of the nictitating membrane of one rabbit. In the new CLP regulation, the criteria to serious eye damage was reversed to "discoloration of the cornea", hence, this dye substance has no longer to be classified to be hazardeous to eyes according to REGULATION (EC) No 1272/2008.

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., OECD Guideline 405 and OPPTS 870.2400 in compliance with GLP.

 

One animal received 100 mg of an undiluted test substance into 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. At 24 and 72 h and after 7, 14 and 21 d, the eyes were further examined for cornea/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.

One hour after administration the conjunctiva of the animal showed slight swelling (including nictitating membrane) and some blood vessels were definitely hyperaemic (injected). Additionally, serous discharge discolored by the test compound with moistening of the lids and hairs just adjacent to the lids was noted. 24 h up to 48 h after administration the conjunctiva of the animal still showed some blood vessels which were definitely hyperaemic (injected). 72 h after administration a slight swelling of the conjunctiva was observed again. 24 h up to 21d after administration the nictitating membrane was discolored by the test compound. This discoloration was not reversible and the study was terminated at Day 21 after administration.

 

Under the test conditions, the test substance showed potential risk of serious damage to the rabbit eye due to the persistent discoloration of the eye according to Council Directive 67/548/EEC.

With the new CLP regulation (Regulation (EC) No 1272/2008), the susbtance has to be evaluated as not irritating to eyes.

The classification according to Council Directive 67/548/EEC and according to Regulation (EC) No 1272/2008 differs due to changes in the criteria for eye irritation in the new CLP regulation. The substance was classifies as "Xi Risk phrases: R41" due to persistent (21 days) discoloration of the nictitating membrane of one rabbit. In the new CLP regulation, the criteria to serious eye damage was reversed to "discoloration of the cornea", hence, this dye substance has no longer to be classified to be hazardeous to eyes according to REGULATION (EC) No 1272/2008.