Methyl 3-aminocrotonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4 November 1999 - 1 December 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Feb. 1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 1992

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

August 1998

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid

Details on test material:

Expiry date: 21 May 2000
Batch: 624

Specific details on test material used for the study:

The test material, a crystalline solid, was stored in the dark at ambient temperature.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Three male young adult New Zealand White rabbits were used. They were within the weight range 2.10-2.34 kg on arrival, and were supplied by Harlan
UK. They arrived at lnveresk on 20 October 1999.The animals were allowed to acclimatise for at least 5 days prior to commencement of the study.
The animals were housed individually in stainless cages (approximate dimensions 77 x 70 x 48 cm) with a Noryl moulded dual level interior and
perforated floor beneath which was an absorbent paper lined tray.
Each cage was supplied with an automatic watering valve and a food hopper. Mean environmental maximum and minimum temperatures were 20°C and
19°C and mean relative humidity was 44%. A 12 h light/dark cycle was in operation (light hours 0700-1900 h) with a minimum of 15 air changes per hour.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

40.1 mg, 39.2 mg and 39.5 mg

Duration of treatment / exposure:

1, 24, 48 and 72 h and extended up to day 28 (to evaluate reversibility). Eyes were not rinsed.

Observation period (in vivo):

Examinations after 1, 24, 48 and 72 h and extended up to day 28 (to evaluate reversibility)

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

Both eyes of each rabbit were examined prior to dosing and no ocular defects were detected.
The requisite quantity of finely ground test material was instilled into the right eye using the following technique:
The test material was placed into the right eye by gently pulling the lower eyelid away from the eyeball to form a sac into which the test material
was dropped. The lids were then gently held together for 1-2 s. The left eye remained untreated to serve as a control.
One rabbit was treated first and observed for signs of severe irritation for 6 days after instillation. As none were noted, the remaining 2 rabbits were
treated in the same way. All the animals were checked for viability early in the morning and again as late as possible on each day. The eyes were examined using a hand held magnifier and pen torch 1, 24,
48 and 72 h after instillation of the test material. Observations were extended up to Day 28, in order to fully evaluate the reversibility of reactions.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.66

Max. score:

3.66

Reversibility:

not fully reversible within: 28 day

Remarks:

still grade 1

Remarks on result:

positive indication of irritation

Remarks:

stained with fluorescein

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 9 days

Remarks on result:

probability of moderate irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 9 days

Remarks on result:

probability of moderate irritation

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 6 days

Remarks on result:

probability of moderate irritation

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

0.33

Reversibility:

fully reversible within: 48 hours

Remarks:

stained with fluorescein

Remarks on result:

probability of weak irritation

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

0.33

Reversibility:

fully reversible within: 48 hours

Remarks on result:

probability of weak irritation

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

1.33

Reversibility:

fully reversible within: 6 days

Remarks on result:

probability of moderate irritation

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.66

Max. score:

0.66

Reversibility:

fully reversible within: 72 hours

Remarks on result:

probability of mild irritation

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.66

Max. score:

1.66

Reversibility:

fully reversible within: 72 hours

Remarks:

stained with fluorescein

Remarks on result:

probability of mild irritation

Results 10

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.66

Max. score:

0.66

Reversibility:

fully reversible within: 72 hours

Remarks on result:

probability of mild irritation

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

1.33

Reversibility:

fully reversible within: 9 days

Remarks on result:

probability of moderate irritation

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 6 days

Remarks on result:

probability of moderate irritation

Irritant / corrosive response data:

One animal (Animal 1) reacted more severely than the other two. An easily discernible translucent area of the cornea (score 2) was noted from 1 h after instillation of the test material, up to Day 28.
The area of opacity covered between a half and the whole surface of the cornea (score 3-4) up to 72 h, and covered a quarter of the cornea (score 1) up to Day 28. Iridial responses and
slight to moderate conjunctival redness were noted up to Day 6. Slight to moderate conjunctival chemosis and moderate discharge were also noted up to 72 h after instillation.
In the 2 remaining animals, scattered or diffuse areas of opacity were noted up to 48 h after instillation, covering up to a quarter of the cornea in one animal and up to three quarters in the other (score 2-3).
Iridial responses were noted in both animals up to 48 h after instillation. Slight to moderate conjunctival redness was noted in both animals, up to either 72 h or Day 6. Both animals
displayed very slight to slight conjunctival chemosis up to 72 h after instillation. Slight or moderate discharge was noted up to 24 h after instillation.

Other effects:

A thick white patch covering the lower right third of the cornea was noted in Animal 1 from Day 9 up to Day 28.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Based on the non-reversible effects of the one animal in a valid and reliable GLP and Guideline-conform study a classification with Eye Corr. 1; H318 is required for methyl 3-aminocrotonate.

Executive summary:

In the eye irritation study one animal showed a prolonged and stronger reaction than the other 2. The reactions in the eye of this animal were still existent on day 28 and therefore, require classification with Eye Corr. 1; H318, respectively. All other animals within the study showed slight signs of irritation but no classification would result from this.

Based on the non-reversible effects of the one animal in a valid and reliable GLP and Guideline-conform study a classification with  Eye Corr. 1; H318 is required for methyl 3-aminocrotonate.