Trisodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonatonaphthyl]azo]-2,5-dimethylphenyl]azo]benzene-1,4-disulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1974

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:

according to guideline

Guideline:

other: U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965)

Deviations:

not specified

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

(clinical observations for systemic effects were not done)

GLP compliance:

no

Remarks:

(Pre-GLP study)

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonatonaphthyl]azo]-2,5-dimethylphenyl]azo]benzene-1,4-disulphonate

EC Number:

274-410-5

EC Name:

Trisodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonatonaphthyl]azo]-2,5-dimethylphenyl]azo]benzene-1,4-disulphonate

Cas Number:

70210-13-8

Molecular formula:

C27H22ClN9O9S3.3Na

IUPAC Name:

trisodium 2-{[4-({4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulfonato-1-naphthyl}diazenyl)-2,5-dimethylphenyl]diazenyl}benzene-1,4-disulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Specific details on test material used for the study:

Name: FAT 40075/A
Purity: 74.5 %

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Bred on the premises of the testing laboratory
- Age at study initiation: 3 to 4 months
- Weight at study initiation: Males: 2.33 kg; Females: 2.06 kg
- Housing: Caged singly
- Diet: A commercial irradiated diet (Styles-Oxoid), ad libitum
- Water: Sterile filtered water, ad libitum
- Acclimation period: One week

ENVIRONMENTAL CONDITIONS
- Temperature: 20 ± 1 °C
- Humidity: 50-70 %
- Photoperiod: Animals were exposed to artificial light for 10 h daily from 08: 00 - 18:00 h

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated right eye

Amount / concentration applied:

- Amount(s) applied: 100 mg

Duration of treatment / exposure:

Unwashed eye: unspecified (not rinsed after single instillation)
Washed eye: 30 sec

Observation period (in vivo):

1, 6, 24, 48 and 72 h.

Number of animals or in vitro replicates:

Six animals (four males and two females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
After 30 seconds the test substance was, as far as possible, flushed out of the eyes of three rabbits with warm water for 1 min.

SCORING SYSTEM: The primary eye irritation index represents the sum of the mean values for cornea, iris and conjunctivae.

CORNEA:
A Opacity and degree of density (most dense area scored)
No opacity 0
Scattered or diffuse area, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque, iris invisible 4

B Area of cornea involved
One quarter (or less) but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4

IRIS:
A Values
Normal 0
Folds above normal, congestion, swelling, circumcornea injection (any or all 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:
A Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal 0
Vessels definitely injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

B Chemosis
No swelling 0
Any swelling above normal (includes nictitating membrane) 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

C Discharge
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, and considerable area around the eye 3
(A + B + C) x 2 Maximum possible score = 20

TOOL USED TO ASSESS SCORE: hand-slit lamp

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 10 d

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 6 d

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 6 d

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 6 d

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #4

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 6 d

Results 10

Irritation parameter:

iris score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 11

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #4

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 10

Results 12

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Results 13

Irritation parameter:

cornea opacity score

Basis:

animal: #3, 5 and 6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Test substance was washed 30 sec post application

Results 14

Irritation parameter:

iris score

Basis:

animal: #3, 5 and 6

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: Test substance was washed 30 sec post application

Results 15

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 6 d

Remarks on result:

other: Test substance was washed 30 sec post application

Results 16

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Test substance was washed 30 sec post application

Results 17

Irritation parameter:

conjunctivae score

Basis:

animal #5

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 6 d

Remarks on result:

other: Test substance was washed 30 sec post application

Results 18

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: Test substance was washed 30 sec post application

Results 19

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #6

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 6 d

Remarks on result:

other: Test substance was washed 30 sec post application

Results 20

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Test substance was washed 30 sec post application

Irritant / corrosive response data:

A moderate conjunctival reaction was seen in all animals 1 h after application of the test substance. All conjunctivae were normal by Day 10. A corneal reaction consisting mainly of small areas of opacity and damage to the corneal epithelium was present from 1 h after application of the test substance in 3/6 animals (all unwashed eyes) and in 1/6 (washed eye) from 6 h. In three animals these changes have resolved by Day 2 but in one case changes were present until Day 6 (unwashed eye).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the test substance was considered to be a non-irritant to rabbit eye.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance (at ca. 74.5 % purity) in rabbits according to U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965) which is equivalent to OECD Guideline 405 with deviations. A single dose of 100 mg of test substance was applied into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The untreated eye served as control. The eyes of three animals were rinsed with warm water for 1 minutes after 30 seconds of test substance instillation. Both with (3/6) and without (3/6) washing, ocular reactions were scored at 1, 6, 24, 48 and 72 h and 6 and 10 d post-instillation. Instillation of the test substance elicited conjunctival redness of a mean score of 2 in 2/3 eyes (unwashed) and had resolved within 10 d after instillation. Unwashed eyes (Day 10) returned to normal later than washed (Day 6). Corneal opacity and chemosis was reversible within 72 h. All the scores for Iritis were zero. Conjuctival redness was reversed within 6 days. Under the study conditions, the test substance was considered to be a non-irritant to rabbit eye.