Disodium 3-[[4-amino-9,10-dihydro-9,10-dioxo-3-[sulphonato-4-(1,1,3,3-tetramethylbutyl)phenoxy]-1-anthryl]amino]-2,4,6-trimethylbenzenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March from 13 to 20, 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

May 12, 1981

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Acid Violet 048

IUPAC Name:

Acid Violet 048

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH 4414 Fuellinsdorf / Switzerland.
- Age at study initiation: 14 - 15 weeks.
- Weight at study initiation: 2.1 - 2.5 kg
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system.
- Diet: pelleted standand Kliba 341, Batch 94/84 rabbit maintenance diet ("Kliba", Kringentalmuehle AG, Switzerland), ad Iibitum.
- Water: community tap water from Itingen, ad libitum.
- Acclimation period: four days under laboratory conditions after veterinary exarnination.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 55 ± 10 %
- Air changes: 10-15 air changes per hour.
- Photoperiod: 12 hours artificial fluorescent Iight/12 hours dark.
- Other: at least 8 hours music/Iight period.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

The individual dose volume was 0.1 g per animal.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: eyes were not rinsed.

OBSERVATIONS
Viability / Mortality: daily.
Body weights: pre-test, day 1 and at termination of test on a Mettler PK 16 balance.

TOOL USED TO ASSESS SCORE: eye examinations were made with a slit-Iamp 30 SL (c. Zeiss AG, Zuerich / Switzerland) and a Varta Cliptrix diagnostic-Iamp (A. Riegger, BaseI / Switzerland).

SCORING SYSTEM
The eye of each animal were examined 1-, 24-, 48-, 72 hours and 7 days after administration.
The irritation was assessed according to the OECD Guidelines for testing of Chemicals, Section 4, number 405 "Acute Eye irritation/corrosion " adopted May 12, 1981. The corrosive properties of the test article the treated and the color of eye were described and recorded.

Corneal irritation
Opacity degree of density (denest area used for assessment)
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than sIight dulling of normal luster), details of iris clearly visible 1
EasiIy discernible translucent area, details of iris slightly obscured 2
Nacrous area, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible through the opacity 4

Iridic irritation
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination thereof, iris still reacting to Iight (sluggish reaction is positive) 1
No reaction to light, hemorrhage, gross destruction (any or aII of these) 2

Conjunctival irritation
Redness (refers to palpebrar and bulbar conjunctivae, cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperemic (injected) 1
Diffuse, crimson coIor, individual vessels not easily discernible 2
Diffuse beefy red 3

Chemosis, lids and/or nictating membranes
No sweIIing 0
Any swelling above normal (includes nictating membranes) 1
Obvious swelling with partial eversion of lids 2
SweIIing with Iids about half closed 3
SweIIing with Iids more than half 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:

1.3 - 1.7

Results 2

Irritation parameter:

iris score

Basis:

animal: 2/3

Time point:

24/48/72 h

Score:

1

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

2 - 3

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 2/3

Time point:

24/48/72 h

Score:

< 2

Irritant / corrosive response data:

Test item showed a primary irritation score of 5.6 when applied to the rabbit eye mucosa.
In the area of application a violet discoloration of the corneal, scIera and conjunctivae was observed within 1 hour after treatment. This could be related to effects of the test article.
No corrosion of the cornea was observed at each of the measuring intervals.

INDIVIDUAL FINDINGS
Animal No 326, male
1 hour - Cornea: opacity, whole, discoloured, ventral. Conjunctivae: edema, clear visible; discoloured, severe, including nictating membrane and conjunctival sac; substance remainders, violet mucous. Sclera: not visible. Discharge: severe.
24 hours - Cornea: opacity, whole, discoloured, ventral. Iris: slight reddened. Conjunctivae: reddened, severe; edema severe. Sclera: not visible. Discharge: severe.
48 hours - Cornea: opacity, severe. Iris: slight reddened. Conjunctivae: reddened, severe; edema severe. Sclera: not visible. Discharge: severe.
72 hours - Cornea: opacity, severe. Conjunctivae: reddened, severe; edema severe. Sclera: not visible. Discharge: severe.
7 days - Cornea: opacity, ventral. Iris: reddened, ventral, slight. Conjunctivae: reddened. Discharge: minimal.

Animal No 327, female
1 hour - Cornea: opacity, whole, discoloured, ventral. Conjunctivae: edema, clear visible; discoloured, severe, including nictating membrane and conjunctival sac; substance remainders, violet mucous. Sclera: not visible. Discharge: severe.
24 hours - Cornea: opacity, whole. Iris: reddened. Conjunctivae: reddened; edema. Sclera: reddened. Discharge: severe.
48 hours - Cornea: opacity, whole. Conjunctivae: reddened; edema. Sclera: reddened. Discharge: severe.
72 hours - Cornea: opacity, slight, diffuse. Conjunctivae: reddened; edema slight. Sclera: reddened. Discharge: moderate.
7 days - Cornea: opacity, clear visible, marginal area. Conjunctivae: reddened, slight.

Animal No 328, female
1 hour - Cornea: reddened, slight; edema, clear visible; discoloured including nictating membrane; substance remainders, violet mucous. Discharge: severe.
24 hours - Cornea: opacity, whole. Iris: reddened. Conjunctivae: reddened; edema. Sclera: reddened. Discharge: severe.
48 hours - Cornea: opacity. Iris: reddened, slight. Conjunctivae: reddened, clear visible; edema, slight. Sclera: reddened. Discharge: severe.
72 hours - Cornea: opacity. Iris: reddened, slight. Conjunctivae: reddened, clear visible; edema slight. Sclera: reddened. Discharge: minimal.
7 days - Cornea: opacity, ventral, minimal.

Other effects:

TOXIC SYMIPTONS / MORTALITY
No acute toxic symptoms were observed in the animals during the test period and no mortality occurred.

BODY WEIGHT
The body weight gain of all rabbits was similar.

Any other information on results incl. tables

Eye irritation score

Animal	Reaction	After					Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs	7 days
326 M	Cornea opacity	1	1	2	2	2	1.67
327 F	Cornea opacity	1	1	2	1	1	1.33
328 F	Cornea opacity	0	1	2	2	1	1.67
326 M	Iris	0	1	1	1	1	1.00
327 F	Iris	0	1	0	0	0	0.33
328 F	Iris	0	1	1	1	0	1.00
326 M	Conjunctival redness	0	3	3	3	2	3.00
327 F	Conjunctival redness	0	2	2	2	1	2.00
328 F	Conjunctival redness	1	2	2	2	0	2.00
326 M	Conjunctival chemosis	2	2	3	2	0	2.33
327 F	Conjunctival chemosis	3	2	2	1	0	1.67
328 F	Conjunctival chemosis	2	2	1	1	0	1.33

Applicant's summary and conclusion

Interpretation of results:

other: Eye Irrit 2 (H319), according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Eye 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 g 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, 72 hours and 7 days. The ocular reactions were scored by the method described into the OECD Guidelines for testing of Chemicals, Section 4, number 405 "Acute Eye irritation/corrosion" adopted May 12, 1981.

Under the conditions of the experiment, test item was found to cause a primary irritation score of 5.6 when applied to the rabbit eye mucosa. In the area of application, a violet discoloration sclera and conjunctivae was observed within 1 hour after treatment. This could be related to effect of the test article. No corrosion was observed at each of the measuring intervals.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were higher than 1, but lower than 3 for corneal opacity (i.e. 1 .3 - 1.7) in all the treated animals; equal to 1 for iritis in 2 out of 3 rabbits, equal to 2 in two rabbits and equal to 3 in the third one for conjunctival redness; lower than 2 in 2 out of 3 tested rabbits for conjunctival oedema. Although the experiment was termined after 7 days and some reactions were still visible, there are no resons to suspect the possibility of non-reversibility of the effects over an observation period of normally 21 days: based on the description of the individual findings provided in the report, the severity of the effects resulted to be reduced respect to severity recorded after 72 hours.

In conclusion, a classification in category 2 of CLP Regulation (EC) No 1272/2008 seems to be appropriate.