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

Skin irritation / corrosion

Administrative data

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

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 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2004

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

1998

Deviations:

no

Test guideline 4

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)

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

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.)

Test system

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.

Results and discussion

In vivo

Results

Results 1

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

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 24 hours

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 4

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.

Any other information on results incl. tables

 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

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 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.