OO-tert-butyl monoperoxymaleate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993-01-18 to 1993-05-28

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)

Version / remarks:

Version 24 February 1987

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EEC Directive 84/449/EEC

Version / remarks:

EEC Publication No. L. 251, September 1984

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

OO-tert-butyl monoperoxymaleate

EC Number:

217-691-1

EC Name:

OO-tert-butyl monoperoxymaleate

Cas Number:

1931-62-0

Molecular formula:

C8H12O5

IUPAC Name:

(2Z)-4-(tert-butylperoxy)-4-oxobut-2-enoic acid

Test material form:

solid: particulate/powder

Details on test material:

SOURCE OF TEST MATERIAL
- CAS No.: 1931-62-0
- Bulk Density: 0.48 g/ml
- Source and lot/batch No.of test material: 221190 ex PDD
- Expiration date of the lot/batch: 1993-07-01
- Purity test date: 1993-01-18
- Purity: approx. 98%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: in refrigerator in the dark
- Stability under test conditions: stable

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: the powdery test substance was instilled as delivered by the sponsor

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

SPF Quality

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: approx. 13 weeks
- Weight at study initiation: 2419 - 2732 grams
- Housing: Individually labelled cages with perforated floors and equipped with an automatic drinking system. Standard laboratory conditions
- Diet (e.g. ad libitum): standard laboratory rabbit diet (LKK-20 pellet diameter 4 mm)
- Water (e.g. ad libitum): free access to tap-water diluted with decalcified water
- Acclimation period: 14 days before start of treatment under test conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C
- Humidity (%): 55%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 h dark, 12 h light

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 48 mg

Duration of treatment / exposure:

24 h

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): in day 2 after fluorescein examination, both eyes of all three animals were rinsed with approx. 50 ml tap-water per eye, using a velocity of flow which did not affect the eyes, to remove residual test substance.

SCORING SYSTEM: Scoring system for ocular lesions see chapter "Any other information on materials and methods"

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

21 d

Score:

1

Max. score:

1

Reversibility:

not reversible

Results 5

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 6

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

not fully reversible within: 21 days

Results 7

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

21 d

Score:

2

Max. score:

2

Reversibility:

not reversible

Results 8

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Results 10

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

21 d

Score:

1

Max. score:

1

Reversibility:

not reversible

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: 21 days

Results 12

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 21 days

Results 13

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

21 d

Score:

2

Max. score:

2

Reversibility:

not reversible

Results 14

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Results 15

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Results 16

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

21 d

Score:

1

Max. score:

1

Reversibility:

not reversible

Results 17

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 21 days

Irritant / corrosive response data:

Corneal opacity, varying from grade 1-3, was observed in animals 2 and 3 during the study, which persisted up to termination of the study {day 22). From day 15 onwards pannus formation (neovascularisation of the cornea) was seen in these two animals. In animal 1 slight dulling of normal lustre of the cornea was observed from day 2 until day 8. Treatment of the eyes with 2% fluorescein, 24 hours after instillation, revealed corneal epithelial damage in all animals. At following treatments with fluorescein, signs of recovery were noted in all animals and the corneal epithelial damage had completely resolved within 14 days after instillation in animal 1. In animals 2 and 3 the epithelial damage had not resolved on day 22.
The iridic irritation had resolved within 7 days after instillation in all three animals.
Moderate to severe irritation of the conjunctivae was observed in all animals over the first week. Recovery was noted from day 15 onwards, but the irritation had not completely resolved at day 22. Reduced elasticity of the eyelids, a reduction of the size of the pupil in the treated eye and/or gray/white discoloration of the conjunctivae, as a sign of necrosis, were observed in animals 2 and 3 during the study.
For more detailed information on the individual response please see Table 2 in "Any other information on results.

Other effects:

After the 24 hour observation the eyes were rinsed with water to remove remnants of the test substance. However the test substance could only be partly removed, which might have influenced further effects.
No systemic toxicity were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Table 2: Individual Irritation Scores

Animal No. 1	Eye Irritation Scores Post-Application after
Observation	24 h	48 h	72 h	average score (24, 48 and 72 h)	21 days after administration
Cornea - opacity	0	0	0	0,0	0
Iris	1	1	1	1,0	0
Conjunctivae redness	2	3	3	2,7	1
Conjunctivae chemosis	4	3	3	3,3	0
Animal No. 2	Eye Irritation Scores Post-Application after
Observation	24 h	48 h	72 h	average score (24, 48 and 72 h)	21 days after administration
Cornea - opacity	2	2	2	2,0	2
Iris	1	1	1	1,0	0
Conjunctivae redness	3	3	3	3,0	1
Conjunctivae chemosis	3	2	2	2,3	0
Animal No. 3	Eye Irritation Scores Post-Application after
Observation	24 h	48 h	72 h	average score (24, 48 and 72 h)	21 days after administration
Cornea - opacity	2	1	1	1,3	2
Iris	1	1	1	1,0	0
Conjunctivae redness	3	3	3	3,0	1
Conjunctivae chemosis	3	2	2	2,3	0

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

In an in vivo acute eye irritation/corrosion study in rabbits according to OECD 405, OO-tert-butyl monoperoxymaleate produced corrosive effects.

Executive summary:

In a primary eye irritation study according to OECD 405, 48 mg of OO-tert-butyl monoperoxymaleate (98% purity) was instilled into the conjunctival sac of one eye of three male New Zealand White Rabbits (13 weeks old) for 24 h. Both eyes were washed with tap-water after 24 h of exposure. Animals were observed for 21 days with examination at 1, 24, 48, 72 h and 7, 14 and 21 days after test substance administration. Eye lesions were scored by the method of Draize.

Under the conditions of the present study, a single ocular application of the test item to rabbits produced adverse effects on cornea, iris and conjunctivae in all animals. The corneal injury persisted at termination of the study in two animals. The iridic irritation had resolved within 7 days after instillation in all three animals. The irritation of the conjunctivae had not resolved completely at termination of the study in all three animals. Neither mortalities nor clinical signs of toxicity were observed.

In this study, OO-tert-butyl monoperoxymaleate is corrosive to the eye and does warrant for classification as Eye Dam. Category 1 according to CLP criteria.