Reaction mass of trisodium [m[7-[[4'-[[6-benzamido-1-hydroxy-3-sulpho-2-naphthyl]azo]-3,3'-dihydroxy[1,1'-biphenyl]-4-yl]azo]-8-hydroxynaphthalene-1,6-disulphonato(7-)]]dicuprate(3-) and trisodium [m[4-[[4'-[[6-benzamido-1-hydroxy-3-sulpho-2-naphthyl]azo]-3,3'-dihydroxy[1,1'-biphenyl]-4-yl]azo]-3-hydroxynaphthalene-2,7-disulphonato(7-)]]dicuprate(3-)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Direct Blue 251

IUPAC Name:

Direct Blue 251

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Housing: all animals were housed in single cages.
- Diet: a standard laboratory diet was provided ad libitum. The batch of the diet used for this study was analysed for chemical and microbiological contaminants.
- Water: water was provided ad libitum.
- Acclimation period: all animals were acclimatized to the environment for a period of five days prior to the start of the study.
- Health check: prior to the test, the eyes of each rabbit were examined for pre-existing casual injury.

ENVIRONMENTAL CONDITIONS
- Temperature: the room temperature was maintained within a median of 21 ± 2 °C recorded by a maximum-minimum thermometer.
- Humidity: 30 - 70 %, measured by a hygrometer.
- Air changes: the rate of air exchange was maintained at approximately 15 changes/hour.
- Photoperiod: the light/dark cycle was 12 hours.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg of the test material was placed into the conjunctival sac of the right eyes of the animals.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

Three male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: eyes unwashed.

SCORING SYSTEM
The eyes were inspected after 30 min, 24, 48 and 72 hours.
Reading scores for ocular lesions were made according to the recommendations of the USA Food and Drug officials, published in 1959 (Appraisal of the Safet of Chemicals in Foods, Drugs and Cosmetics).

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

Remarks on result:

no indication of irritation

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: 72 hrs

Results 4

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The irritation score resulted 0.67, thus the substance was considered to be non-irritant.

Any other information on results incl. tables

Individual reaction

Animal	Reaction	30 min	24 hrs	48 hrs	72 hrs	mean 24/48/72 hrs
Animal 1	Corneal opacity	0	0	0	0	0.00
Animal 2	Corneal opacity	0	0	0	0	0.00
Animal 3	Corneal opacity	0	0	0	0	0.00
Animal 1	Iris	0	0	0	0	0.00
Animal 2	Iris	0	0	0	0	0.00
Animal 3	Iris	0	0	0	0	0.00
Animal 1	Conjunctival redness	0	0	0	0	0.00
Animal 2	Conjunctival redness	0	1	0	0	0.33
Animal 3	Conjunctival redness	0	2	1	0	1.00
Animal 1	Conjunctival chemosis	0	0	0	0	0.00
Animal 2	Conjunctival chemosis	0	0	0	0	0.00
Animal 3	Conjunctival chemosis	0	0	0	0	0.00

Animal	Findings		30 min	24 hrs	48 hrs	72 hrs
Animal 1	Cornea	(A) density	0	0	0	0
		(B) area	0	0	0	0
		AxBx5	0	0	0	0
	Iris and aspect	(C)	0	0	0	0
		Cx5	0	0	0	0
	Conjuntiva	(D) secretion	0	0	0	0
		(E) swelling	0	0	0	0
		(F) redness	0	0	0	0
		(D+E+F) x 2	0	0	0	0
Animal 2	Cornea	(A) density	0	0	0	0
		(B) area	0	0	0	0
		AxBx5	0	0	0	0
	Iris and aspect	(C)	0	0	0	0
		Cx5	0	0	0	0
	Conjuntiva	(D) secretion	0	0	0	0
		(E) swelling	0	0	0	0
		(F) redness	0	1	0	0
		(D+E+F) x 2	0	2	0	0
Animal 3	Cornea	(A) density	0	0	0	0
		(B) area	0	0	0	0
		AxBx5	0	0	0	0
	Iris and aspect	(C)	0	0	0	0
		Cx5	0	0	0	0
	Conjuntiva	(D) secretion	0	0	0	0
		(E) swelling	0	0	0	0
		(F) redness	0	2	1	0
		(D+E+F) x 2	0	4	2	0

Applicant's summary and conclusion

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Non-irritant

Executive summary:

Three male rabbits were used to assay the eye irritation potential of the test item. 100 mg of the substance were placed into the conjunctival sac of the right eyes of the animals. Eye were not washed; they were were inspected after 30 min, 24, 48 and 72 hours. Reading scores for ocular lesions were made according to the recommendations of the USA Food and Drug officials, published in 1959 (Appraisal of the Safet of Chemicals in Foods, Drugs and Cosmetics).

The irritation score resulted 0.67, thus the substance was considered to be non-irritant.

Conclusion

Although the criteria for the evaluation of the eye irritation potential followed in the test are different from those reported into the OECD guideline, the scoring system is the same. Because the raw tables are included into the study report, it is possible to re-evaluate the results according to the CLP Regulation (EC) No 1272/2008. 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 the 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.