2-{4-[4-[4-fluoro-6-(2-(2-vinylsulfonylethoxy)ethylamino)-1,3,5-triazin-2-ylamino]phenylazo]phenylazo}naphthalene-4,6,8-trisulfonate, trisodium salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 June 2001 to 16 July 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

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)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

442-230-8

EC Name:

-

Cas Number:

321679-52-1

Molecular formula:

Hill formula: C31 H25 F N9 Na3 O12 S4 CAS formula: C31 H28 F N9 O12 S4 · 3 Na

IUPAC Name:

trisodium 7-(2-{4-[2-(4-{[4-({2-[2-(ethenesulfonyl)ethoxy]ethyl}amino)-6-fluoro-1,3,5-triazin-2-yl]amino}phenyl)diazen-1-yl]phenyl}diazen-1-yl)naphthalene-1,3,5-trisulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): FAT 40'800/A
- Lot/batch No.: WP 2/01
- Purity: approx. 70%
- Stability: Stable under storage conditions
- Expiry date: 14 February 2007
- Storage: room temperature (range of 20 ±3 °C), away from direct sunlight

Specific details on test material used for the study:

Identity: FAT 40'800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Stability of test item: Stable under storage conditions
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne / France
- Age at study initiation: Male: 10-11 weeks, Females: 8-9 weeks
- Weight at study initiation: Male: 2012 g, Female: 1942-2044 g
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood (RCC Ltd., Füllinsdorf) and haysticks for gnawing.
- Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 90/01) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Haysticks (QS no. 56/01) provided by Eberle Nafag AG, CH-9200 Gossau.
- Water : Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: Under laboratory conditions after health examination.

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

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1g

Duration of treatment / exposure:

21 days

Observation period (in vivo):

up to 21 days

Number of animals or in vitro replicates:

3; (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, FAT 40800/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 delivery of the animals to the termination of test.
Clinical signs: Daily from delivery of the animals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation.

- The primary irritation score was calculated by totaling the mean cumulative scores at 24, 48 and 72 hours and then divided by the number of data points.

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

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0.77

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

- Irritation: Application of the test item to healthy rabbit conjunctivae resulted in a primary irritation score of 0.78. The eye reactions (mean values from 24 to 72 hours) consisted of grade 0.00 corneal opacity, grade 0.00 iris lesions, grade 0.78 redness of the conjunctivae and grade 0.00 chemosis of the conjunctivae. No abnormal findings were observed in the cornea or iris of any animal at any reading. Slight reddening of the conjunctivae was noted in all animals 1 hour after treatment and persisted in two animals up to the 72-hour reading and in the other animal up to the 24-hour reading. Slight swelling of the conjunctivae was also observed in all animals 1 hour after treatment A slight watery discharge was evident in all animals at the 1-hour reading. The sclera could not be assessed in one female at the 1-hour reading due to marked orange staining caused by the test item. Slight reddening was however observed in the other two animals at this reading.
- Corrosion: No corrosion of the cornea was observed at any of the reading times.

Other effects:

- Viability/mortality and clinical signs: No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
- Coloration: Slight to marked orange staining produced by the test item was noted in all animals 1 hour after treatment. Slight staining persisted in the male up to the 72-hour reading, in one female up to the 7-day reading and in the other female up to termination on day 21.
- 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 scores

Animal Number	Time point	Cornea	Iris	Conjunctiva
				Redness	Chemosis
22	24 h	0	0	1	0
23		0	0	1	0
24		0	0	1	0
22	48 h	0	0	1	0
23		0	0	0	0
24		0	0	1	0
22	72 h	0	0	1	0
23		0	0	0	0
24		0	0	1	0

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Conclusions:

FAT 40800/A was found to be irritating to rabbit's eyes basis of irreversible light orange staining produced by the test item.

Executive summary:

In a GLP-compliant eye irritation study, performed according to OECD guideline 405, the test substance (0.1g) was placed in the conjunctival sac of the left eye of three New Zealand White rabbits. The eyes were examined for 21 days. 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 application. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion. The eye reactions (mean values from 24 to 72 hours) consisted of grade 0.00 corneal opacity, grade 0.00 iris lesions, grade 0.78 redness of the conjunctivae and grade 0.00 Chemosis of the conjunctivae. No abnormal findings were observed in the cornea or iris of any animal at any reading. Slight reddening of the conjunctivae was noted in all animals 1 hour after treatment and persisted in two animals up to the 72-hour reading and in the other animal up to the 24-hour reading. Slight swelling of the conjunctivae was also observed in all animals 1 hour after treatment. A slight watery discharge was evident in all animals at the 1-hour reading. The sclera could not be assessed in one female at the 1-hour reading due to marked orange staining caused by the test item. Slight reddening was however observed in the other two animals at this reading. Slight to marked orange staining produced by the test item was noted in all animals 1 hour after treatment. Slight staining persisted in the male up to the 72-hour reading, in one female up to day 7 and in the other female up to termination on day 21. No corrosion was observed at any of the measuring intervals.In conclusion, FAT 40800/A was found to be irritating to rabbit's eyes basis of irreversible light orange staining produced by the test item.