Cyclohexene

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 October, 2010

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 438(Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

110-83-3

IUPAC Name:

110-83-3

Constituent 2

Reference substance name:

Cyclohexene

EC Number:

203-807-8

EC Name:

Cyclohexene

Cas Number:

110-83-8

Molecular formula:

C6H10

IUPAC Name:

cyclohexene

Details on test material:

Lot No. 10817K
Purity: 99.83%

Test animals / tissue source

Species:

other: chicken

Test system

Vehicle:

other: Sodium chloride (Salsol solution 0.9%)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
Amount(s) applied (volume with unit): 30μL

Duration of treatment / exposure:

10 seconds

Observation period (in vivo):

The control eye and test eyes were evaluated pre-treatment and at approximately 30, 60, 75, 120, 180 and 240 minutes after the post-treatment rinse. Minor variations within ±5 minutes were considered acceptable.

Number of animals or in vitro replicates:

N/A

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: Maximum corneal swelling at up to 75min (%)

Basis:

mean

Score:

-2

Remarks on result:

other: ICE Class: Ⅰ

Results 2

Irritation parameter:

other: Maximum corneal swelling at up to 240min (%)

Basis:

mean

Score:

-9

Remarks on result:

other: ICE Class: Ⅱ

Results 3

Irritation parameter:

other: Maximum corneal opacity

Basis:

mean

Score:

0.17

Remarks on result:

other: ICE Class: Ⅰ

Results 4

Irritation parameter:

other: Fluorescein retention

Basis:

mean

Score:

0.17

Remarks on result:

other: ICE Class: Ⅰ

Results 5

Irritation parameter:

other: Other observation

Remarks on result:

other: None

Results 6

Irritation parameter:

other: Overall ICE Class

Remarks on result:

other: 2×Ⅰ, 1×Ⅱ

Any other information on results incl. tables

RESULTS OF COTROL TEST

- Positive Control: Trichloroacetic acid 30 (w/v) %

Observation	Value	ICE Class
Mean maximum corneal swelling at up to 75 min	-14%	Severe
Mean maximum corneal swelling at up to 240 min	-21%	Severe
Mean maximum corneal opacity	3.83	Ⅳ
Mean fluorescein retention	3.00	Ⅳ
Other Observations	Immediate cornea opacity score 4. Severe thinning (up to -21%) was observed in all eyes (3/3) at during the observation period.
Overall ICE Class	-	, 2×Ⅳ

The positive control Trichloroacetic acid 30 (w/v) % was classed as severely irritating, GHS Classification: Category 1.

- Negative Control: Sodium chloride

Observation	Value	ICE Class
Mean maximum corneal swelling at up to 75 min	0%	Ⅰ
Mean maximum corneal swelling at up to 240 min	0%	Ⅰ
Mean maximum corneal opacity	0.00	Ⅰ
Mean fluorescein retention	0.00	Ⅰ
Other Observations	None	-
Overall ICE Class	-	3×Ⅰ

The negative control Sodium chloride 0.9% had no significant effects on the chicken eye in this study.

※APPENDIX 1 TABLES OF ICE CLASSIFICATION

Table 2.1 ICE classification criteria for corneal class

Mean Corneal Swelling (%)	ICE Class
0 to 5	Ⅰ
>5 to 12 †	Ⅱ
>12 to 18 ( >75 min after treatment )	Ⅱ
>12 to 18 ( ≦75 min after treatment )	Ⅲ
>18 to 26	Ⅲ
>26 to 32 ( >75 min after treatment )	Ⅲ
>26 to 32 ( ≦75 min after treatment )	Ⅳ
>32	Ⅳ

† Negative numbers -5% to -12% are also given a grade II. Values below -12% are not classified.

Table 2.2 ICE classificartion for opacity

Mean Maximum Opacity Score	ICE Class
0.0 – 0.5	Ⅰ
0.6 – 1.5	Ⅱ
1.6 – 2.5	Ⅲ
2.6 – 4.0	Ⅳ

Table 2.3 ICE classification criteria for mean fluorescein retention

Mean Fluorescein Retention Score at 30 minutes post - treatment	ICE Class
0.0 – 0.5	Ⅰ
0.6 – 1.5	Ⅱ
1.6 – 2.5	Ⅲ
2.6 – 3.0	Ⅳ

※APPENDIX 2 CLASSIFICATION

Table 3.1 Assessment of the general IN VITRO eye irritancy and regulatory GHS classification.

The following table is used to identify the probably eye irritancy potential of test items. In the case where the result indicates Severely Irritating, then the test item can be classified as Severe. In all other cases the probable level of irritancy can be reported, but a regulatory in vivo rabbit eye irritation test is required for regulatory classification and labelling purposes.

Classification	Combinations of the three ICE Classes
A = Not irritating	3×I 2×I, 1×II 2xII, 1xI
B = Slight irritating (GHS category 2B: Mild irritant / causes eye irritation)	3×II 2×II, 1×III 1×I, 1×II, 1×III
C = Moderate irritating (GHS category 2A: Irritant / causes eye irritation)	3×III 2×III, 1×II 2xI, 1xIV 2×III, 1×IV 2×III, 1×I 2×II, 1×IV 1×II, 1×III, 1×IV
D = Severely irritating (GHS category 1: Irreversible effects on the eye / serious damage to the eye)	3×IV 2×IV, 1×III 2×IV, 1×II 2×IV, 1×I1 Immediate cornea opacity score 3 or 4 Severe loosening of epithelium

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information

Conclusions:

In vitro eye irritation study in the Isolated Chicken Eyes model with Cyclohexene, the results indicate that the test item was not irritating.