Methyl N-[4-[(2-bromo-6-chloro-4-nitrophenyl)azo]phenyl]-N-(3-methoxy-3-oxopropyl)-β-alaninate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: ETAD test method 14.10.76

Version / remarks:

Hazleton Manual of Standard Operating Procedures as applied to ETAD test method 14.10.76 from Code of Federal Regulations (HLE protocol no. 524/6)

Deviations:

no

GLP compliance:

no

Remarks:

Study pre-dates GLP

Test material

Test material information

Constituent 1

Reference substance name:

Methyl N-[4-[(2-bromo-6-chloro-4-nitrophenyl)azo]phenyl]-N-(3-methoxy-3-oxopropyl)-β-alaninate

EC Number:

261-874-9

EC Name:

Methyl N-[4-[(2-bromo-6-chloro-4-nitrophenyl)azo]phenyl]-N-(3-methoxy-3-oxopropyl)-β-alaninate

Cas Number:

59709-38-5

Molecular formula:

C20H20BrClN4O6

IUPAC Name:

methyl N-[4-[(2-bromo-6-chloro-4-nitrophenyl)azo]phenyl]-N-(3-methoxy-3-oxopropyl)-β-alaninate

Test material form:

solid: granular

Details on test material:

Disperse Brown 19

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Six New .Zealand White rabbits obtained from Buxted Rabbit Co. Ltd., Great Totease Farm, Buxted, Nr. Uckfield, Sussex were used for the study. At commencement the animals were free from any eye irritation or defects.

Diet
The animals were allowed unlimited access to filtered mains water and food (Standard Rabbit Diet, BP Nutrition (U.K.) Ltd., Stepfield, Witham, Essex) throughout the study.

Environment
The animals were housed individually in grid floor cages in a single room maintained under conditions of ambient humidity and temperature except that a lower temperature of 14°C is automatically imposed.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

72 hours (no washing following installation of test article)

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

6 animals

Details on study design:

The animals were identified by an indelible number marked on the inner surface of the pinna. The test article (100 mg) was placed in one eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test substance was dropped.
The lids were then gently held together for about one second. The other eye, remaining untreated, served as a control. The eyes were not washed following instillation of the test article.
The eyes were examined 1 hour, 24 hours, 48 hours and 72 hours after instillation and the degree of irritation was scored according to: Draize, J.H. (1959), Assoc. of Food and Drug Officials of the U.S., Austin, Texas, "The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics". Where a substance caused irritation within the first three days after administration the ocular reaction was also assessed on the seventh day. This value was not included in the calculation of the irritant score.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

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

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

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.28

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No further details specified in the study report.

Any other information on results incl. tables

Individual total eye irritation scores

	Individual total scores
Rabbit number	24 hours (a)	48 hours (b)	72 hours (c)
347 348 536 540 579 581	2 4 4 6 8 6	2 2 2 2 2 2	0 0 0 0 0 0
Total	30	12	0
Grand total (a + b + c) = 42

 

Irritation score =          a + b + c

                                   --------------- = 2.3

                                          18

 

Individual conjunctival, iridial, corneal irritation scores

Rabbit No.	TEST ARTICLE Y0154/001
	Time	Conjunctivae			Iris	Cornea		(A+B+C) x2	Dx5	(ExF) x5	Total
		A	B	C	D	E	F
347	1 hr 24 hr 48 hr 72 hr 7 days	2 1 1 0 -	2 0 0 0 -	2 0 0 0 -	0 0 0 0 -	0 0 0 0 -	0 0 0 0 -	12 2 2 0 -	0 0 0 0 -	0 0 0 0 -	12 2 2 0 -
348	1 hr 24 hr 48 hr 72 hr 7 days	2 1 1 0 -	2 1 0 0 -	2 0 0 0 -	0 0 0 0 -	0 0 0 0 -	0 0 0 0 -	12 4 2 0 -	0 0 0 0 -	0 0 0 0 -	12 4 2 0 -
536	1 hr 24 hr 48 hr 72 hr 7 days	2 1 1 0 -	2 1 0 0 -	2 0 0 0 -	0 0 0 0 -	0 0 0 0 -	0 0 0 0 -	12 4 2 0 -	0 0 0 0 -	0 0 0 0 -	12 4 2 0 -
540	1 hr 24 hr 48 hr 72 hr 7 days	2 1 1 0 -	1 1 0 0 -	1 1 0 0 -	0 0 0 0 -	0 0 0 0 -	0 0 0 0 -	8 6 2 0 -	0 0 0 0 -	0 0 0 0 -	8 6 2 0 -
579	1 hr 24 hr 48 hr 72 hr 7 days	2 1 1 0 -	2 1 0 0 -	2 2 0 0 -	0 0 0 0 -	0 0 0 0 -	0 0 0 0 -	12 8 2 0 -	0 0 0 0 -	0 0 0 0 -	12 8 2 0 -
581	1 hr 24 hr 48 hr 72 hr 7 days	2 1 1 0 -	2 1 0 0 -	2 1 0 0 -	0 0 0 0 -	0 0 0 0 -	0 0 0 0 -	12 6 2 0 -	0 0 0 0 -	0 0 0 0 -	12 6 2 0 -

- = observations not made

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The substance did not cause any signs of irritation in rabbit eyes (cornea opacity, iris). Very weak signs of irritation (chemosis, conjunctivae) were fully reversible within 48 and 72 h, respectively. The substance is not classifiable according to CLP criteria.

Executive summary:

A study was carried out to determine the eye irritation score of test article in the albino rabbit.

The study was performed in accordance with Standard Operating Procedures as applied to ETAD test method 14.10.76 from Code of Federal Regulations.

 

Six New Zealand White rabbits were used for the study. At commencement the animals were free from any eye irritation or defects.

The test article (100 mg) was placed in one eye of each animal by gently pulling the lower lid away from the eyeball to form a cup into which the test substance was dropped. The lids were then gently held together for about one second. The other eye, remaining untreated, served as a control. The eyes were not washed following instillation of the test article.

The eyes were examined 1 hour, 24 hours, 48 hours and 72 hours after instillation and the degree of irritation was scored.

Where a substance caused irritation within the first three days after administration the ocular reaction was also assessed on the seventh day.

This value was not included in the calculation of the irritant score.

 

Result

An eye irritation score of 2.3 was obtained. The test article is therefore regarded as a non irritant.