2-[(2-methyl-1-oxoallyl)oxy]ethyl acetoacetate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 October - 24 October 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in accordance with international guidelines

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, Eye Irritation (2-1-5), 12 Nohsan No. 8147, Agricultural Production Bureau, November 24 2000

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

other: Colourless to yellowish clear liquid

Details on test material:

- Name of test material (as cited in study report): LZ649
- Physical state: Colourless to yellowish clear liquid
- Expiration date of the lot/batch: 23 November 2014
- Storage condition of test material: Room temperature in the dark, desiccated

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Animals for this study were selected from a stock supply of healthy adult rabbits of the New Zealand White strain. They were in the weight range of 4.12 to 4.23 kg and 37 or 44 weeks of age, prior to treatment (Day 1). All rabbits were acclimatised to the experimental environment for a period of 18 or 23 weeks prior to the start of the study.

Each animal was housed individually in a plastic cage with perforated floors and was offered 125 g of a standard laboratory rabbit diet per day; drinking water was provided ad libitum. The batch of diet used for the study is analysed for nutrients, possible contaminants and micro-organisms likely to be present in the diet and which, if in excess of specified amounts, might have an undesirable
effect on the test system. A dietary supplement of hay was offered during acclimatisation until two days prior to dose instillation, for the remainder of acclimatisation and throughout the study observation period wholemeal bread was offered.

During the acclimatisation and study period the animals were given small soft white untreated wood blocks for environmental enrichment.
Animal room environmental controls were set to maintain temperature within the range 16 to 20°C, and relative humidity within 40 to 70%. These environmental parameters were recorded and the permanent record archived with other departmental raw data. Lighting was controlled by means of a time switch to give 12 hours of artificial light (06:00 to 18:00 GMT) in each 24 hour period.

Each animal was identified by a numbered tag placed through the edge of one ear. This identification was unique within the Department throughout the duration of the study. Each cage was identified by a coloured label displaying the study number and animal number.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

As supplied by the sponsor. A single ocular dose of a volume of 0.1 mL was administered.

Duration of treatment / exposure:

Single instillation into the eye of the rabbit

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3

Details on study design:

The study was designed to assess eye irritation potential of LZ649 following a single instillation into the eye of the rabbit. The test substance may come into contact with the eye during handling or use.
The eyes of each animal were examined prior to instillation of the test substance to ensure that there was no pre-existing corneal damage, iridial inflammation or conjunctival irritation. Each animal was gently restrained. The dose was instilled into the right eye by pulling the lower eyelid away from the eye ball to form a cup into which the test substance was dropped. The eyelids were then gently held together for one second before releasing; the left eye remained untreated.

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:

0

Reversibility:

fully reversible

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.6

Max. score:

1

Reversibility:

fully reversible

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.1

Max. score:

0.3

Reversibility:

fully reversible

Irritant / corrosive response data:

There was no sign of toxicity or ill health in any rabbit during the observation period.

Other effects:

Injection of the conjunctival blood vessels or a crimson-red conjunctival appearance was evident in all animals throughout the first 24 hours after instillation, persisting in two animals for a further 24 hours. Slight discharge was apparent in all animals one hour after instillation, chemosis was evident in one animal 24 hours after instillation. All animals were overtly normal 72 hours after instillation.
Instillation of the test substance gave rise to practically no initial pain response.

Any other information on results incl. tables

Mean values for ocular lesions for EC (Regulation 1272/2008) and GHS classification

24, 48 and 72 hours after instillation of LZ649

Animal number and sex	Corneal Opacity	Iridial lesions	Redness of Conjunctiva	Chemosis
180 F	0.0	0.0	0.3	0.0
181 F	0.0	0.0	1.0	0.3
182 F	0.0	0.0	0.7	0.0

F Female

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The highest total mean score was 4.7 occurring at the 24 hour observation; accordingly under the criteria of Kay and Calandra (1962) LZ649 was classified as “practically non-irritating” to the eye.

LZ649 did not require labelling in accordance with European Commission
regulation 1272/2008.

Executive summary:

The eye irritation study to the rabbit was performed in October 2012 according to OECD Guideline 405 and GLP. There was no sign of toxicity or ill health in any rabbit during the observation period. The highest total mean score was 4.7 occurring at the 24 hour observation; accordingly under the criteria of Kay and Calandra (1962) LZ649 was classified as “practically non-irritating” to the eye.

LZ649 did not require labelling in accordance with European Commission regulation 1272/2008.