Disodium ar,ar'-[(9,10-dihydro-9,10-dioxo-1,4-anthrylene)diimino]bis[(4-chlorophenoxy)benzenesulphonate]

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March from 09 to 20, 1984

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:

1984

Report date:

1984

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted May 12, 1981

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Acid Green 040

IUPAC Name:

Acid Green 040

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 FueIllinsdorf/SwitzerIand.
- Age at study initiation: 14 - 15 weeks.
- Weight at study initiation: 2.4 - 2.8 kg.
- Housing: individually in stainless steel cages equipped with an automatic cleaning and drinking system.
- Diet: pelleted standard KIiba 341, Batch 94/84 rabbit maintenance diet, ad Iibitum.
- Water: community tap water from Itingen, ad libitum.
- Acclimation period: 4 days under laboratory conditions after veterinary examination.

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

Test system

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A single dose was administered to the left eye of each animal, individual dose volume was 0.1 g per animal.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

2 males and 1 female

Details on study design:

TOOL USED TO ASSESS SCORE: eye examinations were made with a slit-lamp 30 SL.

OBSERVATIONS
- Viability / Mortality: daily.
- Body weights: pre-test, day 1 and at termination.

SCORING SYSTEM
The eyes of each animal were examined 1, 24, 48, 72 hours and 7 days after administration.
The irritation was assessed according to the OECD guideline 405 "Acute Eye Irritation/Corrosion " adopted May 12, 1981.
The corrosive properties of the test article and the color of the treated eye were described and recorded.
The following subjective numerical scoring system was used to grade the eye lesions.

CORNEAL IRRITATION
Opacity, degree of density (densest area used for assessment)
No ulceration of opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Nacreous area for details of iris visible, size of pupil banely discernible 3
Opaque cornea, iris not discernible through the opacity 4

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

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrar and bulbar conjunctivae, cornea and iris)
Blood vessels normal 0
Some blood vessels definitely hyperaemic (injected) 1
Diffuse, crimson color, individual vessels not easily discernible 2
Diffuse beefy red 3

Chemosis, lids and/or nictating membranes
No swelling 0
Any swelling above normal (include nictating 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

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 - <= 1.7

Reversibility:

fully reversible within: 7 days

Results 2

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible within: 48 hrs

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

>= 2.3 - <= 3

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 72 hrs

Irritant / corrosive response data:

Test item showed a primary irritation score of 5.0 when applied to the rabbit eye mucosa.
In the area of application a bluish discoloration of the cornea 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.

Other effects:

TOXIC SYMPTONS / 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

MEAN SCORES PER ANIMAL

Animal N. / sex	Reaction	1 hr	24 hrs	48 hrs	72 hrs	7 days	Mean 24/48/72 hrs
323 M	Corneal opacity	1*	1*	1*	1*	0	1.00
324 M	Corneal opacity	2*	2*	1*	1*	0	1.33
325 F	Corneal opacity	1*	2*	2*	1*	0	1.66
323 M	Iris	0	1*	0	0	0	0.33
324 M	Iris	0	1*	0	0	0	0.33
325 F	Iris	0	1*	0	0	0	0.33
323 M	Conjunctival redness	1	3*	3*	3*	1	3.00
324 M	Conjunctival redness	1	3*	3*	2*	0	2.67
325 F	Conjunctival redness	1	3*	2*	2*	0	2.33
323 M	Conjunctival chemosis	2*	2*	1	0	0	1.00
324 M	Conjunctival chemosis	3*	1	1	0	0	0.67
325 F	Conjunctival chemosis	2*	1	1	0	0	0.67

INDIVIDUAL FINDINGS

Animal No 323, male

After 1 hr	Cornea: opacity, slight, whole; discolored. Conjunctivae: reddened, slight, dorsal; edema, clear visible; discolored, including nictating membrane, substance remainders, greenish mucous diffuse. Sclera: not visible. Discharge: severe.
After 24 hrs	Cornea: opacity, whole, diffuse. Iris: red. Conjunctivae: reddened severe; edema. Sclera: reddened, severe. Discharge: moderate to severe.
After 48 hrs	Cornea: opacity ventral. Conjunctivae: reddened severe; edema slight. Sclera: reddened, severe. Discharge: minimal
After 72 hrs	Cornea: opacity ventral, diffuse, hardly visible. Conjunctivae: reddened severe. Sclera: reddened, severe.
After 7 days	Conjunctivae: reddened, conjunctival sac, hardly visible.

Animal No 324, male

After 1 hr	Iris: darkened. Conjunctivae: reddened, slight; edema, severe, discolored including nictating membrane; substance remainders, greenish mucous, diffuse. Sclera: not visible. Discharge: severe.
After 24 hrs	Cornea: opacity, whole. Iris: red. Conjunctivae: reddened, severe, whole; edema, slight. Sclera: reddened, severe. Discharge: severe.
After 48 hrs	Cornea: opacity, whole. Conjunctivae: reddened, severe, whole; edema, slight. Sclera: reddened, severe.
After 72 hrs	Cornea: opacity, whole. Conjunctivae: reddened, clear visible. Sclera: reddened, ventral
After 7 days	No abnormal changes visible.

AnimaI No 325, female

After 1 hr	Cornea: opacity, slight, whole; discolored. Conjunctivae: reddened, slight, dorsal; edema, clear visible; discolored, including nictating mernbrane, substance remainders, greenish rnucous, diffuse. Sclera: not visible. Discharge: severe.
After 24 hrs	Cornea: opacity, whole. Iris: red, slight. Conjunctivae: reddened, severe; edema, slight. Sclera: reddened, slight. Discharge: moderate.
After 48 hrs	Cornea: opacity, diffuse. Conjunctivae: reddened, clear visible; edema slight. Sclera: reddened, clear visible. Discharge: minimal.
After 72 hrs	Cornea: opacity, ventral slight. Conjunctivae: reddened, clear visible. Sclera: reddened, clear visible. Discharge: moderate.
After 7 days	No abnormal changes visible.

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

Test item is able to cause eye irritation.

Executive summary:

The primary eye irritation study was investigate according to the OECD guideline 405. The irritation potential was assessed placing the test item in the conjunctival sac of rabbit eyes. In the area of application a greenish discoloration of the cornea and conjunctivae was observed within 1 hour after treatment. This could be related to effects of the test article. The mean values from gradings at 24, 48 and 72 hours were between 1 and 1.7 for corneal opacity, lower than 1 for iritis, between 2.3 and 3 for conjunctival redness and lower than 2 for conjunctival oedema, in all of the tested animals. No corrosion was observed at each of the measuring intervals.

The substance was found to cause a primary irritation score of 5.0 when applied to the rabbit eye mucosa.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were between 1 - 1.7 for corneal opacity, lower than 1 for iritis, between 2.3 - 3 for both conjunctival redness and lower than 2 for conjunctival oedema, in all of the tested animals. All the reactions were fully recovered within 7 days.

The test item meets the criteria to be classified as capable to causes serious eye irritation (i.e. Eye Irrit. 2, H319), according to the CLP Regulation No (EC 1272/2008).