Trisodium 1-amino-4-[[3-[[4-chloro-6-[(sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-2,4,6-trimethyl-5-sulphonatophenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

data is from NTRL reports

Data source

Reference

Reference Type:

secondary source

Title:

Acute Oral Toxicity (LD50) in rats, Acute Toxicity (MLD) in rabbits, Acute dermal toxicity (LD50) in rabbits, Dermal Irritation test in rabbits, eye irritation test in rabbits and Inhalation toxicity in rats

Author:

NTRL report

Year:

1983

Bibliographic source:

OTS0215154, NTRL report, last updated 1983

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

To determine the primary irritation index of the test chemical when instilled to the eyes of rabbits

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 5-amino-3-[[4'-[(7-amino-1-hydroxy-3-sulphonato-2-naphthyl)azo][1,1'-biphenyl]-4-yl]azo]-4-hydroxynaphthalene-2,7-disulphonate

EC Number:

219-384-8

EC Name:

Trisodium 5-amino-3-[[4'-[(7-amino-1-hydroxy-3-sulphonato-2-naphthyl)azo][1,1'-biphenyl]-4-yl]azo]-4-hydroxynaphthalene-2,7-disulphonate

Cas Number:

2429-73-4

Molecular formula:

C32H24N6O11S3.3Na

IUPAC Name:

trisodium 5-amino-3-[(E)-2-(4-{4-[(E)-2-(7-amino-1-hydroxy-3-sulfonatonaphthalen-2-yl)diazen-1-yl]phenyl}phenyl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate

Test material form:

solid

Details on test material:

- Name of test material (as cited in study report): trisodium 5-amino-3-[(E)-2-(4-{4-[(E)-2-(7-amino-1-hydroxy-3-sulfonatonaphthalen-2-yl)diazen-1-yl]phenyl}phenyl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate- Common Name: Chlorazol Black BH- Molecular formula : C32H21N6Na3O11S3- Molecular weight : 830.717g/mole- Substance type: organic - Physical State: Solid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS- Housing: housed individually in metal cages elevated above the droppings with, feed- Diet (e.g. ad libitum): Purina Rabbit Chow, ad libitum- Water (e.g. ad libitum): water, ad libitum

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1ml (62.0mg)

Duration of treatment / exposure:

The lids were held closed for 1-second for even distribution of the test substance

Observation period (in vivo):

The eyes were examined and graded for irritation and corneal damage at 1, 24, 48 and 72 hours after instillation

Duration of post- treatment incubation (in vitro):

no data available

Number of animals or in vitro replicates:

Initial test 6 and confirmatory test 6

Details on study design:

REMOVAL OF TEST SUBSTANCE- Washing (if done): noSCORING SYSTEM: The eyes were examined and the grade of ocular reaction was recorded at 1, 24, 48 and 72 hours, using the scale for scoring ocular lesions as outlined by Lehman. A.J. Lehman in Appraisal of the Safety of Chemicals in Foods. Drugs and Cosmetics Assoc., Food and Drug Officials of the U.S. The evaluation of the data was done in accordance with Section 191.12 Chapter 1, Title 21, Code of Federal Regulations under the FHSA

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24 h

Score:

10.2

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Remarks:

Initial test

Results 2

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24 h

Score:

30.3

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Remarks:

Confirmatory test

Irritant / corrosive response data:

Initial test: Ocular damage in the form of erythema, chemosis, discharge, iritis (1 animal) and corneal opacity (2 animals) was observed. Minimal to slight conjunctival irritation (6 animals) and corneal opacity (1 animal) persisted through 72 hours post-treatment.A maximum average score of 10.2 was recorded at 24 hoursConfirmatory test: ocular damage in the form of erythema, chemosis, discharge and corneal opacity was observed. Minimal to slight conjunctival irritation (6 anims18) and corneal opacity (5 animals) persisted through 72 hours post-treatment.A maximum average score of 30.3 was recorded at 24 hours.

Any other information on results incl. tables

Table: Individual scores for the acute eye irritation study         

Group I - Initial test

Rabbit number	Structure	Duration
		1 hour*	24 hours	48 hours	72 hours
1	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	8	6	4	4
2	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	10	6	4	4
3	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	8	8	4	4
4	Cornea	0	10	5	10
	Iris	0	5	0	0
	Conjunctivae	10	6	6	6
5	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	8	6	8	6
6	Cornea	0	10	5	5
	Iris	0	0	0	0
	Conjunctivae	10	6	6	0
Average		9.3	10.2	7.0	6.5

Bluish coloration (slight to deep) of the conjunctival sac(particularly the lower portion) and/or the cornea(all or part)was evident in all animals at each observation.

Group II –confirmatory test

Rabbit number	Structure	Duration
		1 hour*	24 hours	48 hours	72 hours
7	Cornea	0	30	30	30
	Iris	0	0	0	0
	Conjunctivae	10*	8	4	4
8	Cornea	0	20	20	20
	Iris	0	0	0	0
	Conjunctivae	12	6	4	4
9	Cornea	0	20	20	20
	Iris	0	0	0	0
	Conjunctivae	10*	6	4	4
10	Cornea	0	45	45	45
	Iris	0	0	0	0
	Conjunctivae	12	6	4	6
11	Cornea	0	5	5	0
	Iris	0	0	0	0
	Conjunctivae	8	6	4	4
12	Cornea	0	20	30	20
	Iris	0	0	0	0
	Conjunctivae	12	10	6	4
Average		10.7	30.3	29.3	26.8

 

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

A maximum average score of 10.2 was recorded at 24 hours in the initial test. This score was not sufficient to classify the test chemical as per F.H.S.A Guidelines, so a confirmatory test was performed using 6 additional rabbits. In the confirmatory test, ocular damage in the form of erythema, chemosis, discharge and corneal opacity was observed. Minimal to slight conjunctival irritation (6 anims18) and corneal opacity (5 animals) persisted through 72 hours post-treatment.A maximum average score of 30.3 was recorded at 24 hours.Based on the observations and score in the confirmatory test, the test chemical can be considered to be irritating to eyes.

Executive summary:

A study was performed in rabbits to determine the ocular irritation potential of the test chemical.

6 New Zealand white rabbits were used for the study. Both eyes of each animal were examined with Fluorescein Sodium Ophthalmic solution U.S.P. 96 hours before testing and only those animals without observable eye defects or irritation were used. 0.1 ml undiluted test chemical was instilled into the conjunctival sac of 6 New Zealand White rabbits. The lids were held closed for 1-second for even distribution of the test substance. The other eye served as an untreated control. The eyes were not washed following instillation of the test material. The animals were immobilized in a suitable animal restrainer 1 hour subsequent to treatment. After the initial examination at 24 hours all eyes were washed with sufficient clean tap water at body temperature to remove the residual test sample. All eyes were further examined by placing one drop of Fluorescein Sodium Ophthalmic solution U.S.P. on the cornea. After 5 minutes the excess fluorescein was flushed out with sufficient tap water. Fluorescein Sodium in 2% aqueous solution when applied topically to the eye is used as an ophthalmic diagnostic aid for the detection of corneal lesions, injured areas of the cornea appear greenish yellow following application of fluorescein. The eyes were examined and the grade of ocular reaction was recorded at 1, 24, 48 and 72 hours, using the scale for scoring ocular lesions as outlined by Lehman. The evaluation of the data was done in accordance with Section 191.12 Chapter 1, Title 21, Code of Federal Regulations under the FHSA.

Since the results from the initial test were unsatisfactory to classify the substance according to F.H.S.A Guidelines, an additional group of 6 rabbits were selected and tested using the same procedure to obtain suitable information to classify the test chemical. The eyes of animals of Group II were examined with fluorescein approximately 4 hours prior to treatment.

A maximum average score of 10.2 was recorded at 24 hours in the initial test. This score was not sufficient to classify the test chemical as per F.H.S.A Guidelines, so a confirmatory test was performed using 6 additional rabbits. In the confirmatory test, ocular damage in the form of erythema, chemosis, discharge and corneal opacity was observed. Minimal to slight conjunctival irritation (6 anims18) and corneal opacity (5 animals) persisted through 72 hours post-treatment.

A maximum average score of 30.3 was recorded at 24 hours.

Based on the observations and score in the confirmatory test, the test chemical can be considered to be irritating to eyes.