Sodium 1-amino-9,10-dihydro-9,10-dioxo-4-[(2,2,4-trimethylcyclohexyl)amino]anthracene-2-sulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 24th to March 06th, 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

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 Blue 278

IUPAC Name:

Acid Blue 278

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, Fuellinsdorf / Switzerland.
- Age at study initiation: 14 - 15 weeks.
- Weight at study initiation: 2.4 - 2.5 kg.
- Housing: individually in steinless steel cage, equipped with automatic cleaning and drinking system.
- Diet: pelleted standard Kilba 341, batch 93/83, rabbit maintenance diet, ad libitum.
- Water: community tap water, 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 hrs artificial light / hrs darkness.
- Other: at least 8 hours of music / light period.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g per animal

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

1 maes and 2 females

Details on study design:

TOOL USED TO ASSESS SCORE: hand-slit lamp 30 SL and a Varta Cliptrix diagnostic-lamp.

OBSERVATIONS
- Viability/mortality: daily
- Body weight: pre-treatment, day 1 and at termination of the test.

SCORING SYSTEM
The rabbits were examined 1, 24, 48, 72 hours and 7 days after application of the test compound. The ocular reactions were scored according to the OECD guideline 405.

Opacity-degree of density (area most dense taken for reading)
No opacity 0
Scattered or diffuse area (other than slight dulling of normal luster) details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Nacros 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 circumcorneal 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 palpebral and bulbar conjunctivae excluding cornea and iris)
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 nictating membranes
No swelling 0
Any swelling above normal (includes nictitating membranes) 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 within: 72 hrs

Results 2

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

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: 48 hrs

Irritant / corrosive response data:

Test substance showed a primary irritation score of 2.1 when applied to the rabbits eye mucosa.
In the area of application a bluish discoloration of the cornea, sclera and conjunctivae was observed within 1 hour after treatrnent. This could be related to effects of the test article
No corrosion of the cornea was observed at each of the measuring intervals..

TOXIC SYMPTOMS / 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 scores

Animal N / sex	Reaction	1 hr	24 hrs	48 hrs	72 hrs	7 days	Mean 24/48/72 hrs
281 M	Cornea opacity	0	1*	1*	0	0	0.67
282 F	Cornea opacity	0	1*	1*	0	0	0.67
283 F	Cornea opacity	1*	1*	1*	0	0	0.67
281 M	Iris	0	0	0	0	0	0.00
282 F	Iris	0	0	0	0	0	0.00
283 F	Iris	0	0	0	0	0	0.00
281 M	Conjunctivae redness	0	1	1	1	0	1.00
282 F	Conjunctivae redness	0	1	1	1	0	1.00
283 F	Conjunctivae redness	0	1	1	1	0	1.00
281 M	Conjunctivae chemosis	2*	1	0	0	0	0.33
282 F	Conjunctivae chemosis	2*	2*	0	0	0	0.67
283 F	Conjunctivae chemosis	2*	1	0	0	0	0.33

Primary irritation score

Time point	Mean scores
1 hr	2.3
24 hrs	3.3
48 hrs	2.0
72 hrs	1.0

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:

A study was performed to assess the irritation of the test material to the eye of the New Zealand White rabbit. The method followed OECD Guidelines No. 405. A single application 0.1 g of the test material was administered to the eye of three rabbits. Ocular effects were recorded 1, 24, 48 and 72 hours and 7 days after administration.

Test substance showed a primary irritation score of 2.1 when applied to the rabbits eye mucosa. In the area of application a bluish discoloration of the cornea, sclera and conjunctivae was observed within 1 hour after treatrnent. This could be related to effects of the test article. No corrosion of the cornea was observed at each of the measuring intervals.

No acute toxic symptoms were observed in the animals during the test period and no mortality occurred. The body weight gain of all rabbits was similar.

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 of three tested animals. Therefore, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC) No 1272/2008.