3,3'-[[4-[(2-chloro-4-nitrophenyl)azo]phenyl]imino]bis[propiononitrile]

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 24 to 27, 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3,3'-[[4-[(2-chloro-4-nitrophenyl)azo]phenyl]imino]bis[propiononitrile]

EC Number:

223-765-4

EC Name:

3,3'-[[4-[(2-chloro-4-nitrophenyl)azo]phenyl]imino]bis[propiononitrile]

Cas Number:

4058-30-4

Molecular formula:

C18H15ClN6O2

IUPAC Name:

3,3'-[[4-[(2-chloro-4-nitrophenyl)azo]phenyl]imino]bis[propiononitrile]

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST SYSTEM
- Species: Albino rabbit, New Zealand White (SPF-Quality).
- Original Source: Broekman Institute, Someren, The Netherlands.
- Selection of animals: healthy animals, not previously used, were selected prior to treatment. Both eyes of each animal were free of interfering lesions, irritation or defects.
- Total number of animals: 3 male rabbits.
- Allocation and age at start of treatment: animals 1298, 1302 and 1303; approx. 14 weeks.
- Identification: ear tag.
- Acclimatisation: at least five days under laboratory conditions after veterinary examination.

HUSBANDRY
- Conditions: standard laboratory conditions. Air-conditioned with 15 air changes per hour and controlled environment with optimal conditions considered as being a temperature of 21°C and a relative humidity of 55%. Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity. Lighting was 12 hours artificial fluorescent light and 12 hours dark per day. Music during the light period.
- Accommodation: individually in labelled cages with perforated floors and equipped with an automatic drinking system (ITL, Bergen, The Netherlands). 0.1 et standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms, Woerden, The Netherlands) approx. 100 g per day.
- Water: free access to tap-water diluted with decalcified water.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

100 mg per animal

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

TREATMENT
On day 1, prior to test substance administration, the animals were physically examined and the eyes were found to be normal. The test substance (100 ± 1 mg per animal) was instilled in the conjunctival sac of one eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control. Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

2

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

2

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

2

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

ca. 0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

IRRITATION
Instillation of the test substance into one eye of each of the three male albino rabbits affected the iris in two animals and the conjunctivae in all three animals. Slight dulling of normal lustre (corneal opacity grade 0) was observed in animal 1302 on day 1 only. Treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals. The iridic irritation had resolved within 24 hours in animals 1298 and 1302.
The irritation of the conjunctivae had resolved within 72 hours after instillation in all animals.
CORROSION
There was no evidence of ocular corrosion.

Other effects:

COLOURATION:
Red staining of the outside of the eyelids, caused by the test substance, was observed in two animals on days 2 and 3.

TOXIC SYMPTOMS/MORTALITY:
No toxic symptoms were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:

other: CLP criteria not met

Conclusions:

Not irritant

Executive summary:

Method:

The eye irritation of the test substance was determined in an irritation test on rabbits according to OECD Guideline 405 and EU Method B.5.

 

Observations:

The test substancewas instilled into one eye of each of three male albino rabbits, followed by four observations at approximately 1, 24, 48 and 72 hours after test substance administration. Under the conditions of this study,the test substanceresulted in adverse effects on the iris in two animals and on the conjunctivae in all three animals. Slight dulling of normal lustre (corneal opacity grade 0) was observed in one animal on day 1. The iridic irritation had resolved within 24 hours and the irritation of the conjunctivae had resolved within 72 hours after instillation in the animals. Ocular corrosion was not observed in any of the rabbits. No signs of systemic intoxication were observed during the study period.

 

Result:

Not irritant.