trisodium 3-[(E)-2-(4-{[4-chloro-6-({2-[(4-{[4-(ethenesulfonyl)phenyl]amino}-6-fluoro-1,3,5-triazin-2-yl)amino]propyl}amino)-1,3,5-triazin-2-yl]amino}-5-sulfonaphthalen-1-yl)diazen-1-yl]naphthalene-1,5-disulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 July 2006 to 10 August 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Remarks:

RCC Ltd, 4452 Itingen, Switzerland

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 study report): FAT 40826/A
- Lot/batch No.: TZ 5604 BOP 01/06
- Expiration date: 1 February 2011
- Purity: Content of organic part (Na-salt): approx. 78%. Oligomers: 13%. Main component: approx. 48%.
- Stability of test item: Stable under storage conditions
- Storage conditions: At room temperature (range of 20 ± 5 °C) in a exsiccator.

Specific details on test material used for the study:

Identity: FAT 40826/A
Batch no.: TZ 5604 BOP 01/06
Expiration date: February 01, 2011
Purity: Content of organic part (Na-salt): approx. 78 %; Oligomers: 13 %; Main component: approx. 48 %
Solubility in water: Approx. >50 g/L at room temperature
Stability in water: Max. 7 days at room temperature
pH: 7.6 (1 g/L)
Aggregate state/physical form at room temperature: Solid (orange powder)
Storage conditions: At room temperature at about 20 °C, away from direct sunlight
Specific instructions: Store in desiccator

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands BV, Kreuzelweg 53, NL-5961 NM Horst / The Netherlands
- Age at treatment: 13-14 weeks
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Füllinsdorf) and haysticks 4642 (batch no. 77/05, Provimi Kliba AG) were provided for gnawing.
- Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 24/06) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- 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.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

21 days

Observation period (in vivo):

up to 21 days

Number of animals or in vitro replicates:

1 male, 2 females

Details on study design:

TREATMENT:
The eyes of the animals were examined one day prior to test item administration. Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of the results were not used in the test. On the day of treatment, 0.1 g of FAT 40826/A was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test item. The right eye remained untreated and served as the reference control. The treated eyes were not rinsed after instillation.

OBSERVATIONS:
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. No necropsy was performed on the animals sacrificed at termination of observation. The animals were killed by intravenous injection of Pentobarbitone into the ear vein at a dose of at least 1 mL/kg body weight and discarded.

SCORING SYSTEM:
The eye reactions was assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004 at approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14, 17, and 21 days after administration. Eye examinations were made with a Varta Cliptrix diagnostic-lamp (Roth AG, CH-4153 Reinach/Switzerland)

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0.88

Max. score:

3

Reversibility:

fully reversible within: 10 days

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0.44

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritant / corrosive response data:

- No abnormal findings were observed in the cornea or iris of any animal at any of the measurement intervals. Slight to moderate reddening of the conjunctivae was noted in all animals at the 1- and 24-hour reading and persisted as slight reddening in two animals until the 72-hour and 7 day reading. Slight swelling (chemosis) of the conjunctivae was observed in one animal at the 1- and 24-hour reading. Slight swelling to marked swelling with half closed lids was also noted in one animal from the 1- to the 48-hour reading. The reddening of the sclerea was not assessable in two animals for the first 24 hours after treatment due to staining produced by the test item and chemosis. When assessable one animal showed a slight reddening of the sclerea at the 48-hour reading. Slight to moderate ocular discharge was seen in two animals at the 1-hour reading and persisted as slight ocular discharge in one animal at the 24-hour reading. No abnormal findings were observed in the treated eye of any animal 10 days after treatment.
- No corrosion of the cornea was observed at any of the reading times.

Other effects:

- Slight to marked red staining was observed in all animals 1 hour after instillation and persisted as slight to marked yellow staining until the 17-day reading in two animals.
- No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
- The body weights of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

Individual eye score:

Animal No.	Time point	Corneal opacity	Area of Corneal Opacity	Iris	Conjunctiva		Sclera
					Redness	Chemosis
1	After 24 h	0	0	0	1	0	0
2		0	0	0	1	1	n.a.
3		0	0	0	2	2	n.a.
1	After 48 h	0	0	0	0	0	0
2		0	0	0	1	0	0
3		0	0	0	1	1	1
1	After 72 h	0	0	0	0	0	0
2		0	0	0	1	0	0
3		0	0	0	1	0	0

  n.a. = not assessable due to staining and/or swelling produced by the test item

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40826/A is considered to be not irritating or corrosive to the rabbit eye.

Executive summary:

A GLP-compliant eye irritation study with FAT 40826/A was investigated according to OECD guideline 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours, as well as 7, 10, 14, 17 and 21 days after test item instillation. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and Chemosis of the conjunctivae, separately, insofar as numerical scores were assessable. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.33, 1.00 and 0.33 for reddening and 0, 0.33 and 1 for Chemosis, respectively. The instillation of FAT 40826/A into the eye resulted in moderate, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and Chemosis. These effects were reversible and were no longer evident 10 days after treatment. Slight to marked red staining was observed in all animals 1 hour after instillation and persisted as slight to marked yellow staining until 17 days after treatment in two animals. No abnormal findings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. Thus, the test item did not induce significant or irreversible damage to the rabbit eye. Based on the study results, FAT 40826/A is considered to be "not irritating" to the rabbit eye.