Reaction mass of [5-methyl-2-[[4-(4-methylanilino)-9,10-dioxo-1-anthryl]amino]phenyl]sulfonyloxypotassium and [5-methyl-2-[[4-(4-methyl-2-potassiooxysulfonyl-anilino)-9,10-dioxo-1-anthryl]amino]phenyl]sulfonyloxypotassium

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

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)

Version / remarks:

1981

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Acid Green 025

IUPAC Name:

Acid Green 025

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: 12 to 16 weeks old.
- Weight at study initiation: 2.36 - 2.83 kg.
- Housing: rabbits were individually housed in suspected metal cages.
- Diet: rabbit diet, A.W. Tindall Ltd, ad libitum.
- Water: drinking water, ad libitum.
- Acclimation period: minimum 5 days.
- Health check: within 24 hours of commencement of the test both eyes of each rabbit were examined for evidence of ocular irritation or defect using an ophthalmoscope. Animals showing evidence of ocular lesions were rejected and replaced.

ENVIRONMENTAL CONDITIONS
- Temperature: 19 - 20 °C
- Relative humidity: 45 - 50 %
- Air changes: approximately 15 changes per hour.
- Photoperiod: lighting was controlled by a time switch to give 12 hours light and 12 hours of darkness.

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

test item was used as supplied

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml (weight 54 mg) of test item was placed into the right eye of each rabbit.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

Three animals

Details on study design:

TOOL USED TO ASSESS SCORE: examination was facilitated by use of a standard ophthalmoscope.

SCORING SYSTEM
Assessment of damage/irritation was made 1, 24, 48 and 72 hours following treatment, according to the numerical evaluation given by Draize J.H., 1959 Association of Food and Drug Officials of the United States, Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics".

Cornea
Opacity: degree of density (area most dense taken for reading)
No opacity: 0
Scattered or diffuse area of opacity (other than slight dulling of normal lustre) ditails of iris clearly visible: 1
Easily discernible translucent area, 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

IRITIS
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 lids 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: lids and/or nictitating membranes.
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

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 24 hrs

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 48 hrs

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No evidence of corneal opacity was noted during the study period. Iridial inflammation was noted in all treated eye one hour after treatment. Minimal conjunctivitis continued to be noted on day one in one treated eye; the remaining two treated eyes were normal. All treated eyes were normal on day two.

Any other information on results incl. tables

Individual and mean scores for cornea, irirs and conjuntivae

Animal	Reaction	1 hrs	24 hrs	48 hrs	72 hrs	Mean 24/48/72 hrs
Animal 1	Cornea	0	0	0	0	0.0
Animal 2	Cornea	0	0	0	0	0.0
Animal 3	Cornea	0	0	0	0	0.0
Animal 1	Iris	0	0	0	0	0.0
Animal 2	Iris	1	0	0	0	0.0
Animal 3	Iris	1	0	0	0	0.0
Animal 1	Conjunctival redness	1S	1	0	0	0.3
Animal 2	Conjunctival redness	1S	0	0	0	0.0
Animal 3	Conjunctival redness	2S	0	0	0	0.0
Animal 1	Conjunctival chemosis	1	0	0	0	0.0
Animal 2	Conjunctival chemosis	1	0	0	0	0.0
Animal 3	Conjunctival chemosis	2	0	0	0	0.0

S: areas of green staining

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Not irritating

Executive summary:

The eye irritation potential of the test substance was investigated according to thhe method and procedures outlined into the OECD guideline 405. 0.1 ml (weight 54 mg) of test item was placed into the right eye of each rabbit; the other eye remaining untreated, served as a control. The eyes were examined and the grade of ocular reaction was recorded at 1, 24, 48 and 72 hours. The ocular reactions were scored by the method described by Draize J.H., 1959 Association of Food and Drug Officials of the United States, Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics".

No evidence of corneal opacity was noted during the study period. Iridial inflammation was noted in all treated eye one hour after treatment. Minimal conjunctivitis continued to be noted on day one in one treated eye; the remaining two treated eyes were normal. All treated eyes were normal on day two.

Conclusion

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