trisodium 3-amino-4-[4-[4-(2-(2-ethenylsulfonylethoxy)ethylamino)-6-fluoro-1,3,5-triazine-2-ylamino]-2-sulfophenylazo]-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:

Experiment start date - 07 April 1999; Experiment completion date - 03 May 1999; Study completion date - 11 May 1999.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

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:

429-240-8

EC Name:

-

Cas Number:

212652-59-0

Molecular formula:

Hill formula: C25 H22 F N8 Na3 O13 S4 CAS formula: C25 H25 F N8 O13 S4 · 3 Na

IUPAC Name:

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

Details on test material:

Nature of substance: powder
Test item name FAT 40574/B
Solid
Approx 75% purity
Batch WP 23/99

Specific details on test material used for the study:

Identity: FAT 40574/B
Batch: WP 23/99
Purity: Approx 75 %
Appearance: Solid, dark-red powder
Storage: At room temperature at about 20 °C
Expiration Date: 08 February 2006

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Age when treated: Females: 14 weeks; Males: 15 weeks
Identification: By unique cage number and corresponding ear number.
Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, with target ranges for room temperature 22 ± 3 °C and relative humidity 40-70 % (values above 70 % during cleaning process possible). Room environment was monitored continuously with hourly recordings. The room was illuminated by fluorescent light on a 12 hour light/dark cycle. Recorded music was played for approximately 8 hours during the light period.
Accommodation: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
Water: Community tap water from Itingen, available ad libitum.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

Single application in the conjunctival sac

Observation period (in vivo):

1, 24, 48 and 72 hours as well as 7, 14 and 21 days after treatment.

Number of animals or in vitro replicates:

3

Details on study design:

SCORING SYSTEM:
CORNEA
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity..................... 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible................ 1
Easily discernible translucent area, details of iris slightly obscured........................... 2
Nacreous area, no details of iris visible, size of pupil barely discernible..................... 3
Opaque cornea, iris not discernible through the opacity........................................... 4

IRIS
Normal..................................... 0
Markedly deepened rugae, congestion, swelling, moderate circumcomeal hyperemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive)........................................... 1
No reaction to light, hemorrhage, gross destruction (any or all of these)............ 2

CONJUNCTIVAE
Redness (refers to most severe reading of palpebral and bulbar conjunctivae when compared with control eye)
Blood vessels normal.......................................................................... 0
Some blood vessels definitely hyperemic (injected)................................ 1
Diffuse, crimson color, individual vessels not easily discernible............... 2
Diffuse beefy red................................................................................. 3

Chemosis: lids and/or nictitating membranes
No swelling.............................................................................. 0
Any swelling above normal (including nictitating membranes)........ 1
Obvious swelling with partial eversion of lids................................. 2
Swelling with lids about half-closed............................................. 3
Swelling with lids more than half-closed....................................... 4

TOOL USED TO ASSESS SCORE: hand-slit lamp

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animals

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 animals

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 animals

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Reversibility of any observed effect: Changes not fully reversible within 21 days

Other effects:

COLORATION: Irreversible pink staining of the nictitating membrane and the sclera was noted in all animals until study termination on day 21.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Pink staining nictitating membrane and the sclera by the test article was observed in all animals until 21 days after treatment. Based on the findings of this study, FAT 40574/B was classified as Eye corrosion - 1.

Executive summary:

The primary irritation potential of the test article was investigated by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. This study was conducted according to OECD test guideline 405 in a GLP-certified laboratory. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours as well as 7, 14 and 21 days after test article 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 24/48/72h eye irritation scores were recorded. In all animals, slight to moderate swelling of the conjunctivae, as well as slight watery discharge were observed one hour after treatment. In two animals, conjunctival reddening was noted after 24 hours and persisted in one animal until 48 hours after treatment. All signs of irritation were reversible after 72 hours. The 24/48/72 h corneal opacity, iris, conjuctivae and chemosis score was 0, 0, 0.33, 0, respectively. Pink staining nictitating membrane and the sclera by the test article was observed in all animals until 21 days after treatment. Based on the findings of this study, FAT 40574/B was classified as Eye corrosion - 1.