Reaction products of 2,4-diaminobenzensulfonic acid and 2,3-dibromopropanoyl chloride, diazotised, subsequently coupled with 4-amino-5-hydroxynaphthalene-2,7-disulfonic acid, reacted with reaction products of 2,4,6-trichloro-1,3,5-triazine and N-ethylaniline, sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Test substance is equal to one component of the UVCB substance under registration and it only slightly differs from the other main identified components. Details on the read-across are available in section 13. Source study has reliability 1.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reactive Red 272 - ELINCS

IUPAC Name:

Reactive Red 272 - ELINCS

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Chemisch-pharmazeutische Fabrik, D-88397 Biberach
- Weight at study initiation: 2300-2450 g
- Housing: individually in metal cages
- Diet: ad libitum standard rabbit pellet - Nafag No.814, Gossau, Switzerland
- Water: drinking water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

not specified

Controls:

no

Amount / concentration applied:

32 mg (equals 0.1ml in volume)

Duration of treatment / exposure:

Up to 7 days

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

The test substance was placed into the conjunctival sac of the left eye of each animal after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of test substance. The right eye remained untreated and served as a control. The animals were checked daily for systemic symptoms and mortality. The ocular reactions were evaluated 1, 24, 48, and 72 hours after the instillation of test substance. In order to determine the reversibility of the eye reactions additional evaluations of the ocular reactions were needed in this study. A slit-lamp was used to facilitate the evaluation.

SCORING SYSTEM: Draize J.H. 1959
Cornea opacity and degree of density (most dense area scored)
0 no opacity
1 scattered or diffuse area, details of iris clearly visible
2 easily discernible translucent areas, details of iris slightly obscured
3 nacrous areas, no details of iris visible, siz eof pupils barely discernible
4 opaque, iris invisible

Iris
0 normal
1 folds above normal, congestion, swelling, circumcornea injection (nay or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive)
2 no reaction to light, hemorrage, gross destruction (any or all of these)

Conjunctivae redness (refers to palpebral and bulbar conjunctiva excluding cornea and iris)
0 vessels normal
1 vessels definitely injected above normal
2 more diffuse, deeper crimson red, individual vessels not easily discernible
3 diffuse beefy red

Conjuctivae chemosis
0 no swelling
1 any swelling above normal (includes nictitating membrane)
2 obvious swelling with partial eversion of lids
3 swelling with lids about half closed
4 swelling with lids about half to completely closed

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

4

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24 hours

Score:

0.66

Max. score:

4

Results 3

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 48 and 72 hours and 7 days

Score:

0

Max. score:

4

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 and 24 hours

Score:

1

Max. score:

2

Results 5

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 48 and 72 hours and 7 days

Score:

0

Max. score:

2

Results 6

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 1 and 24 hours

Score:

2

Max. score:

3

Results 7

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 48 hours

Score:

1.33

Max. score:

3

Results 8

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 72 hours

Score:

1

Max. score:

3

Results 9

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 7 days

Score:

0

Max. score:

3

Results 10

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 1 hour

Score:

2

Max. score:

4

Results 11

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24 hours

Score:

1.67

Max. score:

4

Results 12

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 48 and 72 hours

Score:

1

Max. score:

4

Results 13

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 7 days

Score:

0

Max. score:

4

Other effects:

- One animal vocalised upon instillation.
- At the end of the observation period (day 7) the conjunctiva of all animals was slightly stained by the test article. This slight staining of the conjunctiva was considered not to represent any risk of serious damage to the eyes because of its minimal intensity and the fact that no functional relevant parts of the eye were involved. In the course of the physiological regeneration of the conjunctiva such a slight staining will most probably disappear without permanent lesions.

Any other information on results incl. tables

animal	777/M				923/M				959/M
time / obs	cornea	iris	redness	chemosis	cornea	iris	redness	chemosis	cornea	iris	redness	chemosis
1 h	1	1	2	2	1	1	2	2	1	1	2	2
24 h	0	1	2	2	1	1	2	2	1	1	2	1
48 h	0	0	2	1	0	0	1	1	0	0	1	1
72 h	0	0	1	1	0	0	1	1	0	0	1	1
mean 24-72 h	0	0.33	1.67	1.33	0.33	0.33	1.33	1.33	0.33	0.33	1.33	1
7 d	0	0	0	0	0	0	0	0	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

other: not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

Under experimental conditions, test substance was found to be not irritant or corrosive.

Executive summary:

Method

In a eye irritation study, performed according to OECD guideline 405, test substance (32 mg) was placed in the conjunctival sac of the left eye of 3 New Zealand White rabbits. The eyes were examined for 7 days.

Results

A mean cornea score of 0.66 was observed at 24 hours; iris score was 1 at 24 hours; chemosis scores were 2, 1.33 and 1 at 24, 48 and 72 hours, respectively. Chemosis scores observed were 2, 1.67, and 1 at 1, 24, and up to 72 hours, respectively.

Scores at the other time points were 0. Although irritation was observed in the eye, this was completely reversible within 7 days. Under test conditions, test substance was found to be not irritating or corrosive.