2,2'-[(1-methylethylidene)bis(cyclohexane-4,1-diyloxymethylene)]bisoxirane

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 August 2004 to 23 September 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

1998

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF Testing Guideline of 12 Nousan No. 8147

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[(1-methylethylidene)bis(cyclohexane-4,1-diyloxymethylene)]bisoxirane

EC Number:

236-502-3

EC Name:

2,2'-[(1-methylethylidene)bis(cyclohexane-4,1-diyloxymethylene)]bisoxirane

Cas Number:

13410-58-7

Molecular formula:

C21H36O4

IUPAC Name:

2-({[4-(2-{4-[(oxiran-2-yl)methoxy]cyclohexyl}propan-2-yl)cyclohexyl]oxy}methyl)oxirane

Test material form:

liquid: viscous

Details on test material:

- Appearance: Liquid, viscous / colourless
- Storage: Room temperature

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 4 - 5 months
- Weight at study initiation: 3.10 - 3.13 kg
- Housing: individually housed in stainless steel wire mesh cages with grating, floor area: 3000 cm²
- Diet: about 130 g/animal per day
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 24°C
- Humidity: 30 - 70%
- Photoperiod: 12 h light (6.00 a.m. - 6.00 p.m.) 12 h darkness (6.00 p.m. - 6.00 a.m.)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 mL

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 animals (one female, two males)

Details on study design:

REMOVAL OF TEST MATERIAL
- About 24 hours after application of the liquid test material the treated eye of the animal(s) was rinsed with 3 to 6 mL of hand warm tap water for 1 to 2 minutes using a syringe with a blunt probe.

SCORING SYSTEM:
- Readings were made at approx. 1, 24, 48 and 72 h after application.

- Assessment of ocular reactions:
Cornea opacity (op):
Degree of density (the most dense area is taken for reading)
0 = No ulceration or opacity
1 = Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 = Easily discernible translucent area, details of iris slightly obscured
3 = Nacreous area, no details of iris visible, size of pupil barely discernible
4 = Opaque cornea, iris not discernible through the opacity
Area of cornea involved (ar):
1 = > 0 ≤ ¼
2 = > ¼ < ½
3 = > ½ < ¾
4 = > ¾

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

Conjunctivae redness (red):
(Refers to palpebral and bulbar conjunctivae, not to cornea and iris)
0 = Blood vessels normal
1 = Some blood vessels definitely hyperaemic (injected)
2 = Diffuse, crimson colour, individual vessels not easily discernible
3 = Diffuse beefy red

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

Discharge (di):
0 = No discharge
1 = Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 = Discharge with moistening of the lids and hairs just adjacent to lids
3 = Discharge with moistening of the lids and hairs, and considerable area around the eye

EVALUATION OF RESULTS
- For evaluation, the calculation of the mean values of corneal opacity, iris, conjunctival redness and chemosis for readings 24, 48 and 72 hours were quoted. Additionally, the reversibility of the observed findings was taken into account.

TOOL USED TO ASSESS SCORE: Daylight tubes "Lumilux" were used for illumination when reading the eye.

OTHER OBSERVATIONS
- Animals were checked for dead/moribund animals twice each workday (beginning and end) and once animals: on Saturdays, Sundays and on public holidays.
- Body weights were recorded just before application of the test material and after the last reading.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.1

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

- Moderate conjunctival redness (grade 2) was observed in all animals after 1 hour and decreased to slight conjunctival redness (grade 1) in one animal after 24 hours. Slight conjunctival chemosis (grade 1) was noted in two animals and slight discharge (grade 1) was observed in all animals one hour after application only.
- In addition, injected scleral vessels in a circumscribed area or circular were observed in all animals at the 1-hour reading.
- The ocular reactions were reversible in two animals within 24 hours and in one animal within 48 hours after application.
- Mean scores calculated for each animal over 24, 48 and 72 hours were 0 .0 for corneal opacity and for iris lesions, 0.0, 0.3 and 0.0 for redness of the conjunctiva and 0.0 for chemosis.

Any other information on results incl. tables

Table 1: Individual ocular examinations and mean values

Readings	Animal	Cornea		Iris	Conjunctiva			Additional findings
		Opacity	Area of cornea involved		Redness	Chemosis	Discharge
1 h	1	0	0	0	2	0	1	*
	2	0	0	0	2	1	1	#
	3	0	0	0	2	1	1	#
24 h	1	0	0	0	0	0	0
	2	0	0	0	1	0	0
	3	0	0	0	0	0	0
48 h	1	0	0	0	0	0	0
	2	0	0	0	0	0	0
	3	0	0	0	0	0	0
72 h	1	0	0	0	0	0	0
	2	0	0	0	0	0	0
	3	0	0	0	0	0	0
Mean 24-72 h	1	0.0	-	0.0	0.0	0.0	-
	2	0.0	-	0.0	0.3	0.0	-
	3	0.0	-	0.0	0.0	0.0	-
Mean	-	0.0	-	0.0	0.1	0.0	-

 * scleral vessels injected, circumscribed area

# scleral vessels injected, circular

Applicant's summary and conclusion

Interpretation of results:

other: Not classified in accordance with EU Criteria

Conclusions:

Under the conditions of this study, the test material is not irritating to the eye.

Executive summary:

The potential of the test material to cause irritation to the eye was investigated in accordance with the standardised guidelines OECD 405, EU Method B5, OPPTS 870.2400 and JMAFF, under GLP conditions.

During the study, 0.1 mL of test material was applied to one eye of three White New Zealand rabbits (stepwise procedure starting with one animal and supplementing two additional animals). About 24 hours after application the eye was rinsed with tap water. The ocular reactions were assessed approximately 1, 24, 48 and 72 hours after application.

Slight or moderate conjunctival redness, slight conjunctival chemosis and slight discharge were observed in the animals up to 24 hours after application at latest. In addition, injected scleral vessels in a circumscribed area or circular were noted in all animals 1 hour after application. The average score (24 to 72 hours) for irritation was calculated to be 0.0 for corneal opacity and iris, 0 .1 for conjunctival redness and 0.0 for chemosis.

Under the conditions of this study, the test material was concluded to be not irritating to eyes.