A mixture of: 4-(2,2,3-trimethylcyclopent-3-en-1-yl)-1-methyl-2-oxabicyclo[2.2.2]octane; 1-(2,2,3-trimethylcyclopent-3-en-1-yl)-5-methyl-6-oxabicyclo[3.2.1]octane; spiro[cyclohex-3-en-1-yl-[(4,5,6,6a-tetrahydro-3,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]furan]; spiro[cyclohex-3-en-1-yl-[4,5,6,6a-tetrahydro-4,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]]furan]

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 19 April 1993 and 3 May 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

A mixture of: 4-(2,2,3-trimethylcyclopent-3-en-1-yl)-1-methyl-2-oxabicyclo[2.2.2]octane; 1-(2,2,3-trimethylcyclopent-3-en-1-yl)-5-methyl-6-oxabicyclo[3.2.1]octane; spiro[cyclohex-3-en-1-yl-[(4,5,6,6a-tetrahydro-3,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]furan]; spiro[cyclohex-3-en-1-yl-[4,5,6,6a-tetrahydro-4,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]]furan]

EC Number:

422-040-1

EC Name:

A mixture of: 4-(2,2,3-trimethylcyclopent-3-en-1-yl)-1-methyl-2-oxabicyclo[2.2.2]octane; 1-(2,2,3-trimethylcyclopent-3-en-1-yl)-5-methyl-6-oxabicyclo[3.2.1]octane; spiro[cyclohex-3-en-1-yl-[(4,5,6,6a-tetrahydro-3,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]furan]; spiro[cyclohex-3-en-1-yl-[4,5,6,6a-tetrahydro-4,6',6',6'a-tetramethyl)-1,3'(3'aH)-[2H]cyclopenta[b]]furan]

Cas Number:

426218-78-2

Molecular formula:

C16H26O

IUPAC Name:

1-methyl-4-(2,2,3-trimethylcyclopent-3-en-1-yl)-2-oxabicyclo[2.2.2]octane; 3,6',6',6'a-tetramethyl-2',3'a,4',5',6',6'a-hexahydrospiro[cyclohexane-1,3'-cyclopenta[b]fura]-3-ene; 4,6',6',6'a-tetramethyl-2',3'a,4',5',6',6'a-hexahydrospiro[cyclohexane-1,3'-cyclopenta[b]fura]-3-ene; 5-methyl-1-(2,2,3-trimethylcyclopent-3-en-1-yl)-6-oxabicyclo[3.2.1]octane

Test material form:

liquid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Three healthy adult rabbits of the New Zealand White strain were obtained from Froxfield (U .K.) Ltd., Petersfield, Hampshire, England.
The animals were in the weight range of 2.7 to 3.5 kg and approximately 11 to 15 weeks of age, prior to treatment (Day 1) . All rabbits were acclimatised to the experimental environment.
The rabbits were selected without conscious bias for the study. They were housed individually in metal cages with perforated floors in Building R14 Room 5 .
A standard laboratory diet SDS Stanrab (P) Rabbit Diet and drinking water were provided ad libitum.
The batch of diet used for the study was not analysed for nutrients, contaminants or micro-organisms.
Results of routine chemical examination of drinking water at source as conducted usually weekly by the supplier, are made available to Huntingdon Research Centre Ltd . as quarterly summaries .
Animal room temperature was maintained at approximately 19°C and relative humidity at 30 - 70% .
These environmental parameters were recorded daily . Air exchange was maintained at approximately 19 air changes per hour and lighting was controlled by means of a time switch to give 12 hours of artificial light (0700 - 1900 hours) in each 24 hours period.
Each animal was identified by a numbered aluminium tag placed through the edge of one ear . This number was unique within the HRC Industrial Toxicology Department throughout the duration of the study . Each cage, was identified by a coloured label displaying the study schedule number, animal
number and initials of the Study Director and Home Office licensee .

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A 0.1 ml amount of the test substance was placed into the lower everted lid of one eye of each animal.

Duration of treatment / exposure:

Up to 1 hour

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3

Details on study design:

TEST SUBSTANCE PREPARATION
The substance was administered as supplied by the Sponsor. The absorption of the substance was not determined. The homogeneity, stability and purity of the substance were the responsibility of the Sponsor .

TREATMENT PROCEDURE
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 or conjunctival inflammation.
One animal was treated in advance of the others, to ensure that if a severe response was produced, no further animals would be exposed (pilot animal see Table 1).
A 0.1 ml amount of the test substance was placed into the lower everted lid of one eye of each animal.
The eyelids were then gently held together for one second before releasing. The contralateral eye remained untreated.

OBSERVATIONS
Clinical signs: All animals were observed daily for signs of ill health or toxicity.
Ocular responses: Examination of the eyes was made after 1 hour and 1, 2, 3 (equivalent to 24, 48 and 72 hours after instillation), 4, and 7 days after instillation for two animals and 1 hour and 1,2,3,4, 7 and 14 days after instillation for the remaining animal. Observation of the eyes was aided by the use of a handheld light.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

1791 (female)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

1789 (female)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

1790 (female)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

1791 (female)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

1789 (female)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

1790 (female)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

1791 (female)

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Injected blood vessels on nictating membrane

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Remarks:

1789 (female)

Time point:

24/48 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Remarks:

1790 (female)

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

1791 (female)

Time point:

24/48/72 h

Score:

2.3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

1789 (female)

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

1790 (female)

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

OCULAR RESPONSES
The numerical values given to the ocular reactions elicited by the substance are shown in Table 1 ('Any other informations on results incl. tables').
No corneal damage or iridial inflammation was observed.
A diffuse crimson colouration of the conjunctiva accompanied by slight to obvious swelling was observed one hour after instillation. Ocular reactions had increased in severity one day after instillation, with a diffuse crimson to beefy red colouration with obvious to marked swelling being observed.
Reactions subsequently ameliorated in one animal, resolving fully three days after instillation, but persisted in the two remaining animals with blood vessels of the nictating membrane appearing injected in one animal three days. after instillation.
The eyes of these two remaining animals were normal 7 to 14 days after instillation.

Other effects:

CLINICAL SIGNS
There were no signs of toxicity or ill health in any rabbit during the observation period.

Any other information on results incl. tables

Table 1                       Ocular Reactions observed after instillation of the substance

 

Rabbit no. and sex	Region of eye		One hour
				1	2	3	4	7	14
1791 female*	Cornea		0	0	0	0	0	0	0
	Iris		0	0	0	0	0	0	0
	Conjunctiva	Redness	2	2	2	2^	2^	2^	0
		Chemosis	2	3	2	2	2	2	0
1789 female	Cornea		0	0	0	0	0	0	-
	Iris		0	0	0	0	0	0	-
	Conjunctiva	Redness	2	2	2	0	0	0	-
		Chemosis	1	2	1	0	0	0	-
1790 female	Cornea		0	0	0	0	0	0	-
	Iris		0	0	0	0	0	0	-
	Conjunctiva	Redness	2	3	3	2	2	0	-
		Chemosis	2	3	3	3	2	0	-

 

* Pilot animal

^ Injected blood vessels on nictating membrane

Applicant's summary and conclusion

Interpretation of results:

other: Eye irritant

Remarks:

in accordance with EU CLP (EC no 1272/2008 and its amendments)

Conclusions:

The substance is an eye irritant in an in vivo acute eye irritation study (OECD TG 405). Based on this result, the substance shall be classified as eye irritation category 2, using the phrase: Causes serious eye irritation.

Executive summary:

The substance was tested in an in vivo acute eye irritation study (OECD TG 405). Three female rabbits were exposed to the substance for a duration of 1 hour using a single application of 0.1 mL. The treatment was followed by an observation period of 14 days. No corneal damage or iridial inflammation was observed. Redness and chemosis were observed (scores 2, 2 and 3 for redness and scores 2, 3 and 3 for chemosis) increasing in severity from 1 hour to 3 days in one animal and from 1 hour to 7 -14 days after installation in the other two animals. There were no signs of toxicity or ill health in any rabbit during the observation period. Based on these results, the substance causes serious eye irritation.