2-(3,5-dimethylhex-3-en-2-yloxy)-2-methylpropyl cyclopropanecarboxylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 25 June 2008 to 03 July 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

According to the OECD Guideline 405 and in compliance with GLP practices.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

August 1998

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Adopted April 24, 2002

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

April 29, 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss legislation on Good Laboratory Practice (signed in September 04, 2008)

Test material

Test material information

Constituent 1

Reference substance name:

2-{[(2R)-3,5-dimethylhex-3-en-2-yl]oxy}-2-methylpropyl cyclopropanecarboxylate; 2-{[(2S)-3,5-dimethylhex-3-en-2-yl]oxy}-2-methylpropyl cyclopropanecarboxylate

EC Number:

700-118-9

Cas Number:

676532-44-8

Molecular formula:

C16H28O3

IUPAC Name:

2-{[(2R)-3,5-dimethylhex-3-en-2-yl]oxy}-2-methylpropyl cyclopropanecarboxylate; 2-{[(2S)-3,5-dimethylhex-3-en-2-yl]oxy}-2-methylpropyl cyclopropanecarboxylate

Test material form:

liquid

Details on test material:

- Description: Colourless liquid
- Formula: C16H28O3
- Molecular weight: 268,4 g/mol

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands BV, Kreuzelweg 53, NL-5961 NM Horst / The Netherlands Postbus 6174
- Age at study initiation: 16-17 weeks (male); 14 weeks (females)
- Weight at study initiation: 2.6, 2.4 and 2.4 kg
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with autoclaved wood (RCC Ltd, Füllinsdorf) and haysticks 4642 for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch nos. 03/08 and 19/08) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 7 days under laboratory conditions after health examination.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23°C
- Humidity (%): 30-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
- Music was played during the light cycle.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

1, 24, 48 and 72 hours after instillation of test material.

Number of animals or in vitro replicates:

1 male and 2 females

Details on study design:

On the day of treatment, the undiluted test substance was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test article. The right eye remained untreated and served as the reference control.

REMOVAL OF TEST SUBSTANCE
- Washing: not reported

OBSERVATION TIME POINTS
- Viability/Mortality and clinical signs: Daily from acclimatization of the animals to the termination of test.
- Body weights: At start of acclimatization, on the day of application and at termination of observation.
- Clinical signs: Daily from acclimatization of the animals to the termination of test.
- Ocular reaction: approx 1, 24, 48 and 72 hours after the instillation.

SCORING SYSTEM: The eye reactions were assessed according to the following Draize score. Scleral reddening and ocular discharge were also assessed.

GRADING OF OCULAR LESIONS
CORNEA

Opacity: degree of density (area most dense taken for reading):
No ulceration or opacity = 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible = 1
Easily discernible translucent area, details of iris slightly obscured = 2
Nacreous area, no details of iris visible, size of pupil barely discernible = 3
Opaque cornea, iris not discernible through the opacity = 4

Area of cornea involved:
Zero = 0
One quarter (or less) but not zero = 1
Greater than one quarter, but less than half = 2
Greater than half, but less than three quarters = 3
Greater than three quarters, up to whole area = 4

IRIS
Normal = 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive) = 1
No reaction to light, hemorrhage, gross destruction (any or all of these) = 2

CONJUNCTIVAE
Redness (refers to most severe reading of palpebral and bulbar conjunctivae when compared with control eye)
Blood vessels normal = 0
Some blood vessels, definitely hyperemic (injected) = 1
Diffuse, crimson color, individual vessels not easily discernible = 2
Diffuse beefy red = 3

Chemosis: lids and/or nictitating membranes
No swelling = 0
Any swelling above normal (including nictitating membranes) = 1
Obvious swelling with partial eversion of lids = 2
Swelling with lids about half-closed = 3
Swelling with lids more than half-closed = 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

animal: #2 and #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Remarks on result:

probability of mild irritation

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

iris score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

chemosis score

Basis:

animal: #1, #2 and #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00, 0.67 and 0.67 for reddening and 0.00 for chemosis for all three animals.

No abnormal findings were observed in the cornea of iris of any animal at any of the measurement intervals.

A slight to moderate reddening of the conjunctivae was noted at the 1-hour reading and persisted as slight in two animals up to 48 hours after treatment. A slight swelling (chemosis) of the conjunctivae was observed in one animal 1 hour after treatement.

No abnormal findings were observed in the treated eye of any animal 72 hours after treatment, the end of the observation period for all animals.

Other effects:

No staining produced by the test item of the treated eye was observed.
No corrosion of the cornea was observed at any of the reading times.
The body weight of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

Individual findings:

Time after exposure	Rabbit No.	Sex	Corneal opacity	Iris	Conjunctivae		Sclera
					Redness	Chemosis
1 hour	69	M	0	0	1	0	0
	70	F	0	0	1	0	0
	71	F	0	0	2	1	0
24h	69	M	0	0	0	0	0
	70	F	0	0	1	0	0
	71	F	0	0	1	0	0
48h	69	M	0	0	0	0	0
	70	F	0	0	1	0	0
	71	F	0	0	1	0	0
72h	69	M	0	0	0	0	0
	70	F	0	0	0	0	0
	71	F	0	0	0	0	0

 

Mean score at 24, 48 and 72h after a single exposure of 0.1 mL

 

Animal number	Sex	Corneal opacity	N	Iris	N	Conjunctivae
						Redness	N	Chemosis	N
69	M	0.00	3	0.00	3	0.00	3	0	3
70	F	0.00	3	0.00	3	0.67	3	0	3
71	F	0.00	3	0.00	3	0.67	3	0	3

N=number of available data points.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, test substance is not classified as irritating to eyes according to the criteria of the Annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

Executive summary:

In an eye irritation study performed according to the OECD Guideline No. 405, and in compliance with GLP, 0.1 mL of undiluted test material was instilled into one eye of one male and two females New Zealand White rabbits. The other eye remained untreated and served as control. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h after treatment and graded according to the Draize method.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00, 0.67 and 0.67 for reddening and 0.00 for chemosis for all three animals.

No abnormal findings were observed in the cornea of iris of any animal at any of the measurement intervals. A slight to moderate reddening of the conjunctivae was noted at the 1-hour reading and persisted as slight in two animals up to 48 hours after treatment. A slight swelling (chemosis) of the conjunctivae was observed in one animal 1 hour after treatement.

No abnormal findings were observed in the treated eye of any animal 72 hours after treatment, the end of the observation period for all animals.

No staining produced by the test item of the treated eye was observed.
No corrosion of the cornea was observed at any of the reading times.
The body weight of all rabbits were considered to be within the normal range of variability.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0 / 0 / 0 for cornea score; 0 / 0 / 0 for iris score; 0 / 0.67 / 0.67 for conjunctivae score and 0 / 0 / 0 for chemosis score.

Under the test conditions, test substance is not classified as irritating to eyes according to the criteria of the Annex I of the Regulation EC No. 1272/2008 (CLP) and to the GHS. This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.