tetrasodium 3-amino-4-{[4-({4-fluoro-6-[phenyl(2-{[2-(sulfonatooxy)ethyl]sulfonyl}ethyl)amino]-1,3,5-triazin-2-yl}amino)-2-sulfonatophenyl]diazenyl}-5-hydroxynaphthalene-2,7-disulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From October 12, 2004 to November 02, 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

457-630-8

EC Name:

-

Cas Number:

2138836-26-5

Molecular formula:

Hill formula: C29H23FN8Na4O16S5

IUPAC Name:

tetrasodium 3-amino-4-[(1E)-2-[4-({4-fluoro-6-[phenyl({2-[2-(sulfonatooxy)ethanesulfonyl]ethyl})amino]-1,3,5-triazin-2-yl}amino)-2-sulfonatophenyl]diazen-1-yl]-5-hydroxynaphthalene-2,7-disulfonate

Test material form:

solid: particulate/powder

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 Kißlegg
- Body weight at treatment: 2.8 kg
- Housing: In fully air-conditioned rooms in separate cages arranged in a battery
- Diet: Ssnif K-H (V2333), ad libitum and hay (i.e., approximately 15 g daily)
- Water: Tap water in plastic bottles, ad libitum
- Acclimation period: 1 week under study conditions
- Animal identification: Ear tags

ENVIRONMENTAL CONDITIONS
- Temperature: 18±3°C (short lasting deviations are permissible, e.g., during cleaning processes)
- Humidity: 50±20% (short lasting deviations are permissible, e.g., during cleaning processes)
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: October 12, 2004 to November 02, 2004

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48 , 72 h and after 7, 14 and 21 d following administration of the test substance

Number of animals or in vitro replicates:

1 female

Details on study design:

ROUTE OF ADMINISTRATION: Conjunctival

FREQUENCY OF ADMINISTRATIONS: Single dose

PREPARATION OF THE TEST SUBSTANCE:
The animal was treated with 0.1 g (i.e., 100 mg) of test substance

TEST PROCEDURE
An initial test was performed using one animal.

As the test substance showed potential risk of serious damage to the eye only one animal was used in the study.

About 24 h before the start of the study both eyes of the animal was examined under UV light for corneal lesions after instillation of one drop of a 0.01% fluorescein-sodium solution. Only animals without ocular abnormalities were used for the study.

0.1 g of test substance was administered once to the conjunctival sac of the left eye of one rabbit. In each case the untreated eye served as a control.

24 h after administration the treated eye was washed out thoroughly with isotonic saline at approximately 37°C. The eye was also washed out at designated examinationtimes at which discharge was observed or a corneal examination with fluorescein was performed.

The eye was 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 eye 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. All other changes or toxic effects were recorded when apparent. Since effects were still present in the eye after 72 h, further examinations were carried out after 7, 14 and 21 d.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Max. score:

3

Remarks on result:

not determinable

Remarks:

Due to the violet discoloration of the eye the animal could not be assessed for redness of conjunctiva

Irritant / corrosive response data:

From 1 h up to 1 d after application the animal showed slight swellings of lids. The irritations were attended by eye discharge discolored by the test substance (i.e., violet) 1 h after administration. Additionally, conjunctiva, nictitating membrane and iris were discolored by the test substance (i.e., violet) up to the end of the study (i.e., Day 22).

Other effects:

No clinical signs of systemic toxicity were observed.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the substance caused violet discoloration of the conjunctiva, nictating membrane and iris of one rabbit eye until end of the study period. However, no signs of irritation was found and the substance is therefore not considered irritating to the eye according to CLP criteria.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in female NZW White (Crl:KBL(NZW) rabbits according to OECD Guideline 405 and EU Method B.5, in compliance with GLP.

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of one animal. The treated eye was washed out 24 h after administration. The untreated eye served as a control.

The conjunctiva, nictitating membrane and iris were discolored (violet) by the test substance up to the end of the study (Day 22). Overall mean scores for eye irritation were calculated to be 0.0 for iris and opacity of cornea and 0.33 for chemosis of conjunctiva. However, due to the violet discoloration of the eye, the animal could not be assessed for redness of conjunctiva. No clinical signs of systemic toxicity were seen.

Under the test conditions, the substance caused violet discoloration of the conjunctiva, nictating membrane and iris of one rabbit eye until end of the study period.