5-[(4-chloro-2-nitrophenyl)azo]-1-ethyl-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Start of experiment: November 7, 1989; End of experiment: November 14, 1989; Study completion date: 14 November 1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

.

Deviations:

no

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

5-[(4-chloro-2-nitrophenyl)azo]-1-ethyl-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile

EC Number:

274-660-5

EC Name:

5-[(4-chloro-2-nitrophenyl)azo]-1-ethyl-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile

Cas Number:

70528-90-4

Molecular formula:

C15H12ClN5O4

IUPAC Name:

5-[(4-chloro-2-nitrophenyl)diazenyl]-1-ethyl-6-hydroxy-4-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

Test material form:

not specified

Specific details on test material used for the study:

Test article: FAT 36019/F
Batch No.: EN 162190.96
Storage conditions: room temperature
Validity: October 1994
Safety precautions: gloves and face masks
Test material received: October 26, 1999

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

The albino rabbit is the recommended species for Acute Eye Irritation/Corrosion studies.
- Animal strain: New Zealand white rabbits (Chbb:NZW)
- Breeder: Dr. K. Thomae GMBH Chemisch-pharmazeutische Fabrik D-7950 Biberach/Riss
- Acclimatisation period: at least 5 days-

Group size and husbandry:
The test was performed on 3 female rabbits, checked for normal eye conditions, weighing between 2630 to 2860 g (approx. 9-13 weeks old) . The animals were housed individually in metal cages, identified by ear tattoo, kept at a constant room temperature of 20+3 °C, at a relative humidity of 30-70 % and on a 12 hours light cycle day. The rabbits received ad libitum standard rabbit pellet - Nafag No. 814, Gossau, Switzerland - and fresh water.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml (42 mg)

Duration of treatment / exposure:

one single application

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 female rabbits

Details on study design:

0.1 ml (42 mg) of FAT 36091/F 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 the test article. 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 FAT 36091/F. A slit-lamp was used to facilitate the evaluation.

SCORING SYSTEM:
CORNEA
- Opacity: degree of density (area most dense taken for reading).
- No ulceration or opacity 0
- Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible 1
- Easily discernible translucent area, details of iris slightly obscured 2
- Nacrous area, no details of iris visible, size of pupil barely discernible 3
- Opaque cornea, iris not discernible through the opacity 4

IRIS
- Normal 0
- Markedly deepended rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive) 1
- No reaction to light, haemorrhage, gross destruction ( any or all of these ) 2

CONJUNCTIVAE
Redness: (refers to palpebral and bulbar conjunctivae, cornea and iris)
- Blood vessels normal 0
- Some blood vessels definitely hyperaemic (injected) 1
- Diffuse, crimson colour, individual vessels not easily discernible 2
- Diffusy beefy red 3

CHEMOSIS
lids and/or nictating membranes
- No swelling 0
- Any swelling above normal (includes nictating membranes ) 1
- Obvious swelling with partial eversion of lids 2
- Swelling with lids about half closed 3
- Swelling with lids more than, half closed 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.78

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Other effects:

No abnormal increase in body weight was noted.

Any other information on results incl. tables

Individual eye scores

	CORNEA		IRIS		CONJUNCTIVA
Animal number 310/F					Redness	Chemosis
	ce	te	ce	te	ce/te	ce/te
After 1 hour	0	0	0	0	0/2	0/1
After 24 hours	0	0	0	0	0/1	0/0
After 48 hours	0	0	0	0	0/1	0/0
After 72 hours	0	0	0	0	0/1	0/0
Mean 24 -72 hours	0	0	0	0	0/1	0/0
After 7 days	0	0	0	0	0/0	0/0

	CORNEA		IRIS		CONJUNCTIVA
Animal number 304/F					Redness	Chemosis
	ce	te	ce	te	ce/te	ce/te
After 1 hour	0	1	0	0	0/2	0/2
After 24 hours	0	0	0	0	0/1	0/0
After 48 hours	0	0	0	0	0/0	0/0
After 72 hours	0	0	0	0	0/0	0/0
Mean 24 -72 hours	0	0	0	0	0/0.33	0/0
After 7 days	0	0	0	0	0/0	0/0

	CORNEA		IRIS		CONJUNCTIVA
Animal number 318/F					Redness	Chemosis
	ce	te	ce	te	ce/te	ce/te
After 1 hour	0	0	0	0	0/2	0/2
After 24 hours	0	0	0	0	0/1	0/0
After 48 hours	0	0	0	0	0/1	0/0
After 72 hours	0	0	0	0	0/1	0/0
Mean 24 -72 hours	0	0	0	0	0/1	0/0
After 7 days	0	0	0	0	0/0	0/0

ce=control eye

te= test eye

M=Male

F=Female

Body weights (q)

Animal N°	310/F	404/F	318/F
At start of test	2860	2630	2670
After 3 days	2870	2690	2840
After 7 days (end)	2970	2760	2980

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions employed FAT 36091/F induced irritation of the cornea and conjunctiva when instilled into the conjunctival sac of albino rabbits.

Executive summary:

The eye irritation in New Zaeland White after single application was performed to determined the irritant potency of the test item on the rabbit eyes. This test was based on the OECD Guideline No. 405, adopted May 12, 1981, by the OECD council, and on Annex V, part B of Council Directive 79/831/EEC (Commission Directive 84/449/EEC of April 25, 1984; Official Journal L251, September 19, 1984). 0.1 ml (42 mg) of FAT 36091/F 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 the test article. The right eye remained untreated and served as a control. The animals were checked daily for systemic symptoms and mortality. Because reactions were observed within 72 hours after instillation of the test article, the observation period was extended to 7 days to determine the reversibility of the eye reactions. No effects were observed on corneal opacity, iris and chemosis. Mean 24/48/72h conjuctiva score was found to be 0.78. According to the EEC classification of the results obtained 24/48.72 hours after instillation FAT 36091/F can be classified as non-irritant to albino rabbit's eye. The eye reactions observed were reversible within 72 hours.