Reaction mass of (3R,3aS,6R,7R,8aS)-3,6,8,8-tetramethyloctahydro-1H-3a,7-methanoazulen-6-yl rel-acetate and rel-(1aR,4aR,8aR)-2,4a,8,8-tetramethyl-1,1a,4,4a,5,6,7,8-octahydrocyclopropa[d]naphthalene and rel-(3R,3aS,7S,8aS)-3,6,8,8-tetramethyl-2,3,4,7,8,8a-hexahydro-1H-3a,7-methanoazulene

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 April 2001 - 16 May 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The in vivo test was performed before the REACH regulation came into force, requesting in vitro studies. This information is used for read across to Cedryl Acetate EOA

Data source

Reference

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

17 July 1992

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

31 July 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

[1S-(1α,3aβ,4α,8aβ)]-decahydro-4,8,8-trimethyl-9-methylene-1,4-methanoazulene

EC Number:

207-491-2

EC Name:

[1S-(1α,3aβ,4α,8aβ)]-decahydro-4,8,8-trimethyl-9-methylene-1,4-methanoazulene

Cas Number:

475-20-7

Molecular formula:

C15H24

IUPAC Name:

(1S,3aR,4S,8aS)-4,8,8-trimethyl-9-methylidenedecahydro-1,4-methanoazulene

Test material form:

liquid

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne, France
- Age at study initiation: 11 weeks
- Weight at study initiation: 2.1 - 2.2 kg
- Housing: Individually in stainless steel cages.
- Diet: Free access to pelleted standard Provimi Kliba 3418 rabbit maintenance diet (Provimi Kliba AG, CH-4303 Kaiseraugst).
- Water: Free access to community tap water from Füllinsdorf
- Acclimation period: no data.

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 20 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 14
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: Adjacent areas of the untreated skin of each animal served as controls.

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

14 days

Number of animals:

1 male and 2 females

Details on study design:

TEST SITE:
Four days before treatment, the left flank was was clipped with an electric clipper, exposing an area of approximately 100 square centimeters (10x10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was again clipped.

TREATMENT
On the day of treatment, 0.5 mL of the test substance was places on a surgical gauze patch (ca. 2.5 cm x 2.5 cm). This gauze patch was applied to approx. 6 square centimeters of the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and enchored with tape.

REMOVAL OF TEST SUBSTANCE
After 4 hours the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site.

OBSERVATIONS
- Mortality/Viability and Clinical signs: Daily from delivery of the animals to the termination of the test.
- Body Weight: At the start of acclimatization, on the day of application and at termination of observation.
- Irritation: The skin reactions were assessed at approximately 1, 24, 48 and 72 hours and at 7, 10 and 14 days after the removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded.

SCORING SYSTEM:
The irritation was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992.
If evident, corrosive or staining properties of the test substance were described and recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.33

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

At 1 hour, in one animal moderate erythema was seen; at the same time in 2/3 severe swelling was seen. At 24/48/72 hours well-defined erythema were seen in all animals with maximum score 2; at this time point 2/3 animals showed moderate to severe swelling with average score of 2.33 and one animal scored 1.33. All skin reactions were clear 14 days after treatment.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
Scaling was evident at the test site of all animals from 7 to 10 days after treatment. In addition, the skin of one female was noted to be dry and inelastic at these readings.
No staining by the test substance of the treated skin was observed. No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.
The body weights of all animals were considered to be within the normal range of variability.

Applicant's summary and conclusion

Interpretation of results:

other: Skin irritant Category 2

Remarks:

according to EU CLP (EC No. 1272/2008 and its amendments).

Conclusions:

In this skin irritation study with rabbits, performed according to OECD 404 guideline and GLP principles, moderate erythema (score 2) were seen and moderate to severe edema (score 1-3) were seen at 24/48 and 72hours. Two out of three animals have an average edema score of = 2.33, while the other animal scored an average of 1.33. The irritant effects were reversible within 14 days. This means that the substance is a skin irritant.

Executive summary:

The substance was tested for 4 hours in a skin irritation test in 3 rabbits according to OECD TG 404 test guideline and GLP principles.

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred. In this study moderate erythema (score 2) were seen and moderate to severe edema (score 1 -3) were seen at 24/48 and 72hours. Two out of three animals have an average edema score of = 2.33, while the other animal scored average of 1.33 for edema. The irritant effects were reversible within 14 days. This means that the substance is a skin irritant.