Reaction mass of Cobaltate(3-), bis[2-[[[3-[2-[1-[[(2-chlorophenyl)amino]carbonyl]-2-(oxo-κO)propyl]diazenyl-κN1]-4-(hydroxy-κO)phenyl]sulfonyl]amino]benzoato(3-)]-, sodium (1:3) and tetrasodium bis[2-[[[3-[[1-[(2-chloroanilino)carbonyl]-2-oxopropyl]azo]-4-hydroxyphenyl]sulphonyl]amino]benzoato(3-)]cobaltate(4-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1974

Report date:

1974

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

(observation period was up to 7 d instead of 21 d; eyes were not examined at 1 h after test substance application; clinical observations for systemic effects was not done)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: the technique described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US Association of Food and Drug Officials (AFDO).

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Trisodium bis[2-({[3-({1-[(2-chlorophenyl)amino]-3-(hydroxy-kO)-1-oxobut-2-en-2-yl}diazenyl-kN1)-4-(hydroxy-kO)phenyl]sulfonyl}amino)benzoato(3-)]cobaltate(3-) and Trisodium [2-({[3-({1-[(2-chlorophenyl)amino]-3-(hydroxy-kO)-1-oxobut-2-en-2-yl}diazenyl-kN1)-4-(hydroxy-kO)phenyl]sulfonyl}amino)benzoato(3-)][2-({[3-({1-[(2-chlorophenyl)amino]-3-(hydroxy-kO)-1-oxobut-2-en-2-yl}diazenyl-kN2)-4-(hydroxy-kO)phenyl]sulfonyl}amino)benzoato(3-)]cobaltate(3-)

EC Number:

941-792-6

Cas Number:

not given

Molecular formula:

C46H32Cl2CoN8O14S2.3Na

IUPAC Name:

Reaction mass of Trisodium bis[2-({[3-({1-[(2-chlorophenyl)amino]-3-(hydroxy-kO)-1-oxobut-2-en-2-yl}diazenyl-kN1)-4-(hydroxy-kO)phenyl]sulfonyl}amino)benzoato(3-)]cobaltate(3-) and Trisodium [2-({[3-({1-[(2-chlorophenyl)amino]-3-(hydroxy-kO)-1-oxobut-2-en-2-yl}diazenyl-kN1)-4-(hydroxy-kO)phenyl]sulfonyl}amino)benzoato(3-)][2-({[3-({1-[(2-chlorophenyl)amino]-3-(hydroxy-kO)-1-oxobut-2-en-2-yl}diazenyl-kN2)-4-(hydroxy-kO)phenyl]sulfonyl}amino)benzoato(3-)]cobaltate(3-)

Test material form:

not specified

Details on test material:

- Name of test material (as cited in study report): FAT 20049/A
- Substance type: Dye
- Analytical purity: Approx. 89%
- Lot/batch No.: EN 14086

Specific details on test material used for the study:

Test Item: FAT 20049/A
Purity: approx. 89 %

Test animals / tissue source

Species:

rabbit

Strain:

other: Russian breed

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Housing: The rabbits were caged singly in V2A wire-mesh cages
- Diet: NAFAG, Gossau SG, rabbit food (ad libitum)
- Water: ad libitum
- Only rabbits showing normal ophthalmic findings were included in the test.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the right eye served as control

Amount / concentration applied:

- Amount applied: 100 mg
- Concentration: Undiluted

Duration of treatment / exposure:

The treated eyes of three (out of six) rabbits were each rinsed with 10 ml lukewarm water about 30 seconds after treatment.

Observation period (in vivo):

7 d

Number of animals or in vitro replicates:

6 (i.e., 3 males and 3 females)

Details on study design:

- The test substance in an amount of 100 mg was inserted into the conjunctival sac of the left eye of the rabbits and the eyelids were held open for a few seconds. The right eye was not treated and served as an untreated control. In 3 of the 6 rabbits approx. 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water.

- The eye irritation was evaluated with a slit-lamp on 1, 2, 3, 4 and 7 d (i.e., 24, 48, 72, 96 and 168 h).

SCORING SYSTEM:
- The ocular reactions were scored by the method described in Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the AFDO.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

1.9

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Fully reversible within 4 d when rinsed after 30 seconds

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

1.7

Max. score:

3

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Fully reversible within 7 d when rinsed after 30 seconds

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

2.1

Max. score:

4

Reversibility:

not fully reversible within: 7 d

Remarks on result:

other: Fully reversible within 7 d when rinsed after 30 seconds

Irritant / corrosive response data:

- The irritation index was found to be 15.3 for the cornea, 0 for the iris and 9 for the conjunctivae.
- The observed cornea and conjunctivae reactions in the unwashed eyes were not reversible within the observation period of 7 d.

Other effects:

none

Any other information on results incl. tables

Results of the eye irritation:

Eyes washed after application of test substance:

Days after application	Rabbit 1 (F)			Rabbit 2 (F)			Rabbit 3 (F)
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
1	0	0	8	15	0	10	10	0	12
2	0	0	6	5	0	6	10	0	8
3	0	0	6	0	0	0	5	0	6
4	0	0	0	0	0	0	0	0	6
7	0	0	0	0	0	0	0	0	0

 

Unwashed eyes after application of test substance:

 

Days after application	Rabbit 4 (M)			Rabbit 5 (M)			Rabbit 6 (M)
	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva	Cornea	Iris	Conjunctiva
1	30	0	16	20	0	16	20	0	16
2	30	0	14	30	0	14	20	0	14
3	40	0	14	30	0	12	30	0	14
4	40	0	12	30	0	12	30	0	12
7	40	0	12	5	0	12	20	0	12

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The test substance was considered to be irritating to the eyes of rabbits.

Executive summary:

An in vivo study was conducted to evaluate the acute eye irritation potential of the test substance (at ca. 89 % purity) in rabbits according to the procedure described in the Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics (1959) of the US Association of Food and Drug Officials (AFDO) which is similar to the OECD Guideline 405 but with few deviations. 100 mg of the test substance was placed into the conjunctival sac of the left eye of six rabbits (i.e. 3 male and 3 females). The right eye remained untreated and was used as control. In three rabbits, washing was done with 10 mL of lukewarm water after 30 seconds of test substance instillation. Assessment of ocular damage/irritation was made with a slit-lamp at 24, 48, 72, 96 and 168 h following treatment. The irritation index was calculated to be 15.3, 0 and 9 for the cornea, iris and conjunctivae, respectively. The mean scores for corneal opacity, iris, conjunctival redness and chemosis were considered to be 1.9, 0, 1.7 and 2.1. The score obtained for corneal opacity thus fulfils the criteria for classification as eye irritant category 2 as specified by the Regulation (EC) No. 1272/2008. Thus, the test substance was considered irritating to the eyes of rabbits.