Tetrasodium 4,4'-bis[[4-[bis(2-hydroxypropyl)amino]-6-[(4-sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-stilbene-2,2'-disulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 1 to 4, 1986.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium 4,4'-bis[[4-[bis(2-hydroxypropyl)amino]-6-[(4-sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-stilbene-2,2'-disulphonate

EC Number:

267-097-1

EC Name:

Tetrasodium 4,4'-bis[[4-[bis(2-hydroxypropyl)amino]-6-[(4-sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-stilbene-2,2'-disulphonate

Cas Number:

67786-25-8

Molecular formula:

C44H48N12Na4O16S4

IUPAC Name:

tetrasodium 2,2'-ethene-1,2-diylbis[5-({4-[bis(2-hydroxypropyl)amino]-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)benzenesulfonate]

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Nottingham University, School of Agriculture, Sutton Bonington, Leicestershire, U.K.
- Age at study initiation: approx. 12 to 16 weeks old.
- Weight at study initiation: 2.4 - 2.69 kg.
- Housing: animals were individually housed in suspended metal cages.
- Diet: free access, Rabbit Diet (Special Diet Services Limited, witham, Essex, U.K).
- Water: free access, driking water.
- Acclimation period: minimum five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 - 23 °C
- Humidity: 50 - 60 %
- Air changes: approx. 15 changes per hour.
- Photoperiod: 12 hours light and 12 hours darkness.

Test system

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 ml (which was found to weight 94 mg).

Number of animals or in vitro replicates:

Three.

Details on study design:

CONTROL BEFORE STARING
Within 24 hours of commencement of the test both eyes of each rabbit provisionally selected, were examined for evidence of ocular irritation or defect using an ophthalmoscope. Animals showing evidence of ocular lesions were rejected and replaced.

APPLICATION
The test substance was placed into the right eye of each rabbit by gently pulling the lower lid away from the eyeball to form a cup into which the test material was dropped. The upper and lower eyelids were held together for about one second immediately after application to prevent loss of the test material from the eye. The left eye remained untreated and was used for control purposes.
Following treatment of the first rabbit one crop or local anaesthetic (Ophthaine) was instilled into both eyes of the remaining two rabbits approximately 1 - 2 minutes before dosing.

SCORING SYSTEM
The eye irritation was assessed at 24, 48, 72 hours and 4 and 7 days after treatment.

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

IRIS:
A Values
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, 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 palpebral and bulbar conjunctivae excluding cornea and iris)
Blood vessels normal: 0
Some blood vessels definitely hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse beefy red: 3

CHEMOSIS:
No swelling: 0
Any swelling above normal (includes nictitating membrane): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling with lids about half closed to completely closed: 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Results 2

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible

Irritant / corrosive response data:

A dulling of the normal lustre of the corneal surface was noted in one treated eye one hour after treatment; no corneal opacity was noted in this eye at subsequent observations. The cornea of the other treated eyes appeared normal throughout the study period.
Iridial inflammation was noted in two treated eyes one hour after treatment. The iris of the remaining treated eye appeared normal at this time. No iridial inflammation inflammation was noted at the 24, 48 and 72-hour observations.
Conjunctivitis was noted in all treated eyes one and twenty-four hours after treatment. All treated eyes were normal at the 48 and 72-hour observations.

Any other information on results incl. tables

Animal	1 hr	24 hrs	48 hrs	72 hrs	Mean 24, 48, 72 hrs
Cornea
1 (6)	0	0	0	0	0.00
2 (64)	0	0	0	0	0.00
3 (71)	0	0	0	0	0.00
Iritis
1 (6)	1	0	0	0	0.00
2 (64)	1	0	0	0	0.00
3 (71)	0	0	0	0	0.00
Conjunctival - Redness
1 (6)	2	1	0	0	0.33
2 (64)	2	1	0	0	0.33
3 (71)	1	1	0	0	0.33
Conjunctival - Chemosis
1 (6)	1	0	0	0	0.00
2 (64)	1	0	0	0	0.00
3 (71)	1	0	0	0	0.00

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information according to the CLP Regulation Criteria used for interpretation of results: EU

Conclusions:

Not irritating.

Executive summary:

Method

The test was performed on 3 New Zealand White rabbits. The test material in an amount of 0.1 g was inserted into placed into the right eye of the rabbits.

The eye irritation assessment was made at 1, 24, 48 and 72 hours.

Results

A dulling of the normal lustre of the corneal surface was noted in one treated eye one hour after treatment; no corneal opacity was noted in this eye at subsequent observations. The cornea of the other treated eyes appeared normal throughout the study period.

Iridial inflammation was noted in two treated eyes one hour after treatment. The iris of the remaining treated eye appeared normal at this time. No iridial inflammation inflammation was noted at the 24, 48 and 72-hour observations.

Conjunctivitis was noted in all treated eyes one and twenty-four hours after treatment. All treated eyes were normal at the 48 and 72-hour observations.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in all animals.

Therefore the substance does not meet the criteria to be classified as eye irritating.

In conclusion, the test item can be classified as NON irritating, according to the the CLP (EC 1272/2008) Regulation.