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

Endpoint summary

Description of key information

Two in-vivo irritation studies were carried out for the test item. A skin irritation and corrosivity study in the rabbit was performed during December 1984 according to OECD Guideline 404 and GLP. No adverse skin reactions were noted during the study period and no corrosive effects were noted.
A primary irritation index of 0 was obtained. The test item was regarded as non-irritating to rabbit skin.
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.
However, the test item LZ649 is not irritating to the eye and does not require labelling in accordance with European Commission regulation 1272/2008.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

during December 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study conducted in accordance with international guidelines.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Three young adult female New Zealand White rabbits obtained from Ranch Rabbits, Crawley, were used for the study.
At the start of the study the rabbits weighed between 3.82 and 4.65 kg and were at least 12 weeks old. They were acclimatised to the
laboratory environment for at least 3 days and were examined for signs of ill health or injury shortly before the study commenced.
All animals appeared healthy and no animals were rejected.
The animals were individually housed in grid floor cages in a single air conditioned room at a temperature between 19 and 25°C and a
relative humidity between 40 and 70%. The animals were exposed to a constant daily photoperiod of 12 hours artificial light and 12 hours
darkness. Temperature and humidity were recorded twice daily on weekdays and once daily at week-ends.
The animals were allowed free access to mains water and food (Standard Rabbit Diet, Special Diets Services Ltd., Stepfield,) throughout the study.
The animals were individually identified by means of a number on a metal ear tag. Each cage was identified by a coloured card giving details of
HLE project number, test article, dosage, sex, animal number, date of treatment, route of administration and the name of the
Home Office licensee responsible for the study.

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 mL were applied to each treatment site.

Duration of treatment / exposure:

contact period 4 hours

Observation period:

One hour after removal, the evaluation was repeated 24, 48 and 72 hours after removal of the patches.

Number of animals:

3

Details on study design:

The back of each animal was clipped free of hair the day prior to treatment using electric veterinary clippers.
A portion of test article (0.5 ml) was applied to an area of skin approximately 6 cm2 on each animal and covered with a gauze patch.
The patch was occluded and secured using a strip of impermeable adhesive tape. A plastic collar was placed around the neck of each
animal to prevent premature removal of the wrappings and ingestion of the test article.
Four hours after application the patches were removed and the skin wiped with a disposable paper towel moistened with water to remove
any test article still remaining.

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No adverse skin reactions were noted during the study period. No corrosive effects were noted.

Other effects:

Not other effects observed.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

No adverse skin reactions were noted during the study period. A primary irritation index of 0 was obtained. P5306 was regarded as
non-irritating to rabbit skin. No corrosive effects were noted.

Executive summary:

The skin irritation and corrosivity study in the rabbit was performed during December 1984 according to OECD Guideline 404 and GLP. No adverse skin reactions were noted during the study period and no corrosive effects were noted.

A primary irritation index of 0 was obtained. P5306 was regarded as non-irritating to rabbit skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

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

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

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

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)

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.

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.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.6

Max. score:

1

Reversibility:

fully reversible

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.

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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:
GLP study conducted in accordance with international guidelines, reliable without restriction.

Justification for selection of eye irritation endpoint:
GLP study conducted in accordance with international guidelines, reliable without restriction.

Justification for classification or non-classification

Based on the irritation/corrosion data available, the substance is not classified as irritating and does not require labelling according to Regulation 1272/2008/EEC (CLP) and according to Directive 67/548/EEC (DSD).