Reaction mass of Cobaltate(2-), [2-[[[4-hydroxy-3-[[2-oxo-1-[(phenylamino)carbonyl]propyl]azo]phenyl]sulfonyl]amino]benzoato(3-)][2-[[2-hydroxy-5-[(phenylamino)sulfonyl]phenyl]azo]-3-oxo-N-phenylbutanamidato(2-)]-, disodium and Cobaltate(3-), bis[2-[[[4-hydroxy-3-[[2-oxo-1-[(phenylamino)carbonyl]propyl]azo]phenyl]sulfonyl]amino]benzoato(3-)]-, trisodium and sodium bis[2-[[2-hydroxy-5-[(phenylamino)sulphonyl]phenyl]azo]-3-oxo-N-phenylbutyramidato(2-)]cobaltate(1-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November from the 01st to the 08th, 1985

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:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

May 12, 1981

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Acid Yellow 235

IUPAC Name:

Acid Yellow 235

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: 15 - 16 weeks.
- Weight at study initiation: 2.4 - 2.9 kg
- Housing: individually in stainless cages equipped with an automatic cleaning and drinking system.
- Diet: pelleted standard Kliba 341, Batch 10/85 rabbit maintenance diet ("KIiba" Klingentalmuehle AG, Switzerland), ad Iibitum.
- Water: community tap water from Itigen, ad libitum.
- Acclimation period: four days under test conditions after veterinary examination.

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

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Application volume: 0.1 g per animal

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

OBSERIVATIONS
- Viability / mortality: daily.
- Bodyweights: Day 1 and at termination of test on a Mattler PK 16 balance.

TOOL USED TO ASSESS SCORE: slit-lamp 30 SL.

OBSERVATION TIME POINTS
For each individual animal, the irritation scores for erythema and edema were recorded at 60 minutes, 24, 48 and 72 hours after administration; in addition, any lesions, a description of the degree and nature of irritation, corrosion or reversibility and any toxic effect observed were recorded.

SCORING SYSTEM
Cornea Grade
Opacity: degree of density (readings should be taken from most dense area)
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 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

Iris
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia; or injection; iris reactive to light (a sluggish reaction is positive) 1
Hemorrhage, gross destruction, or no reaction to light 2

Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and iris)
Normal 0
Some blood vessels hyperaemic (injected) 1
Diffuse,crimson colour; individual vessels not easily discernible 2
Diffuse beefy red 3

Swelling (refers to lids and/or nictating membranes)
Normal 0
Some swelling above normal 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: 2/3

Time point:

24/48/72 h

Score:

< 1

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

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Irritant / corrosive response data:

Under the conditions of the experiment, the test item was found to cause a primary irritation score of 2.4 when applied to rabbit eye mucosa.
In the area of application a slight yellow discoloration of the cornea and conjunctivae was observed which could be related to effects of the test article.
No corrosion effect had occurred on the skin at each measuring interval.

Other effects:

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

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

Any other information on results incl. tables

Animal N. / sex	Reaction	Observation at				Mean 24/48/72 hrs
		1 hr	24 hrs	48 hrs	72 hrs
295 M	Cornea opacity	1	1	1	0	0.67
296 F	Cornea opacity	1	1	1	1	1.00
297 F	Cornea opacity	0	0	0	0	0.00
295 M	Iris	0	0	0	0	0.00
296 F	Iris	0	0	0	0	0.00
297 F	Iris	0	0	0	0	0.00
295 M	Conjunctival redness	2	1	1	1	1.00
296 F	Conjunctival redness	2	2	1	1	1.33
297 F	Conjunctival redness	2	1	0	0	0.33
295 M	Conjunctival chemosis	3	1	0	0	0.33
296 F	Conjunctival chemosis	2	1	0	0	0.33
297 F	Conjunctival chemosis	1	0	0	0	0.00

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 purpose of the primary eye irritation study was to assess possible irritation potential when single doses of test item are placed in the conjunctival sac of rabbit eyes. The experiment was conducted in accordance with the method and procedures outlined into the OECD guideline 405.

Under the conditions of the experiment, the test item was found to cause a primary irritation score of 2.4 when applied to rabbit eye mucosa. In the area of application a slight yellow discoloration of the cornea and conjunctivae was observed which could be related to effects of the test article.

No corrosion effect had occurred on the skin at each measuring interval.

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 at least 2 out of three tested animals. Thus, the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC) No 1272/2008.