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

Endpoint summary

Description of key information

Eye Irritation (Remmele, 2005)

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

Skin (Remmele, 2005)

Under the conditions of the study, the test material was not irritating to the skin.

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:

19 July 2004 to 23 September 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nousan No. 8147

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 7 - 8 months
- Weight at study initiation: 3.67 - 3.91 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.)

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 mL undiluted test material applied to a 2.5 x 2.5 cm patch

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 animals (2 males and 1 female)

Details on study design:

TEST SITE
- Area of exposure: Flank, dorsolateral part of the trunk
- % coverage: a 2.5 x 2.5 cm patch moistened with the test material was applied to the untreated clipped skin.
- Type of wrap if used: semiocclusive dressing (patch and fleece)

REMOVAL OF TEST SUBSTANCE
- The test material was removed at the end of the exposure period (4 hours) with polyethylenglycol and polyethylenglycol/water (1:1).

OBSERVATION TIME POINTS
- Immediately after removal of the patch, approx. 1, 24, 48 and 72 h after removal of the patch and then in weekly intervals maximally up to day 14.
- Body weights were determined before application of the test material and after the last reading.
- Daylight tubes "Lumilux" were used for illumination for assessing the treatment sites.
- Twice each workday (beginning and end) and once on Saturdays, Sundays and on public holidays the cages were checked for dead/moribund animals.

SCORING SYSTEM:
- Erythema and eschar formation:
No erythema = 0
Very slight erythema (barely perceptible = 1
Well defined erythema = 2
Moderate to severe erythema = 3
Severe erythema (beet redness) to eschar formation preventing grading of erythema = 4

- Oedema formation:
No oedema = 0
Very slight oedema (barely perceptible) = 1
Slight oedema (edges of area well defined by definite raising) = 2
Moderate oedema (raised approx. 1 mm) = 3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) = 4
Description of any dermal findings not covered by this scale were also recorded.

EVALUATION OF RESULTS
- For evaluation, the calculation of the mean values of erythema and oedema for readings at 24, 48 and 72 hours were quoted. Additionally, the reversibility of the observed findings was taken into account.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

- Moderate erythema (grade 2) was observed in all animals immediately after removal of the patch and after 1 hour. Slight erythema (grade 1) was noted in two animals after 24 hours and persisted in one animal up to 72 hours after removal of the patch.
- In one animal erythema was extended beyond the area of exposure up to 1 hour after removal of the patch.
- The cutaneous reactions were reversible within 24 hours, 48 hours or 7 days after removal of the patch in one animal each.
- Mean scores over 24, 48 and 72 hours for each animal were 0.3, 0.0 and 1.0 for erythema and 0.0 for oedema.

 Table 1: Irritant response data

Readings	Animal	Erythema	Oedema	Additional findings
0 h	1	2	0
	2	2	0
	3	2	0	Erythema extending beyond the area of exposure
1 h	1	2	0
	2	2	0
	3	2	0	Erythema extending beyond the area of exposure
24 h	1	1	0
	2	0	0
	3	1	0
48 h	1	0	0
	2	0	0
	3	1	0
72 h	1	0	0	Study discontinued because the animal was free of findings
	2	0	0	Study discontinued because the animal was free of findings
	3	1	0
7 days	3	0	0
Mean 24-72 h	1	0.3	0.0
	2	0.0	0.0
	3	1.0	0.0
Mean	-	0.4	0.0

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 skin.

Executive summary:

The potential of the test material to cause irritation to the skin was determined in accordance with the standardised guidelines OECD 404, EU Method B.4, OPPTS 870.2500 and JMAFF, under GLP conditions.

During the study 0.5 mL of test material was applied for 4 hours to the intact skin of three rabbits, using a patch of 2.5 cm x 2.5 cm, covered with semi-occlusive dressing. After removal of the patch the application area was washed. The cutaneous reactions were assessed immediately after removal of the patch, approximately 1, 24, 48, 72 hours and 7 days after removal of the patch.

Slight or moderate erythema was observed in the animals during the course of the study. In one animal erythema was partly extended beyond the area of exposure. The cutaneous reactions were reversible in all animals within 7 days after removal of the patch at latest. The average score (24 to 72 hours) for irritation was calculated to be 0 .4 for erythema and 0.0 for oedema.

Under the conditions of this study, the test material was not considered to be irritating to the 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:

02 August 2004 to 23 September 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

1998

Deviations:

no

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)

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.)

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.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

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.

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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Eye Irritation (Remmele, 2005)

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. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

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.

Skin (Remmele, 2005)

The potential of the test material to cause irritation to the skin was determined in accordance with the standardised guidelines OECD 404, EU Method B.4, OPPTS 870.2500 and JMAFF, under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

During the study 0.5 mL of test material was applied for 4 hours to the intact skin of three rabbits, using a patch of 2.5 cm x 2.5 cm, covered with semi-occlusive dressing. After removal of the patch the application area was washed. The cutaneous reactions were assessed immediately after removal of the patch, approximately 1, 24, 48, 72 hours and 7 days after removal of the patch.

Slight or moderate erythema was observed in the animals during the course of the study. In one animal erythema was partly extended beyond the area of exposure. The cutaneous reactions were reversible in all animals within 7 days after removal of the patch at latest. The average score (24 to 72 hours) for irritation was calculated to be 0 .4 for erythema and 0.0 for oedema.

Under the conditions of this study, the test material was not considered to be irritating to the skin.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to skin and eye corrosion or irritation.