1,4-Methanonaphthalen-5(1H)-one, 9-(dichloromethylene)-2,3,4,6,7,8-hexahydro-, oxime, (5E)-

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 January 2011 to 16 August 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-[(3E)-11-(dichloromethylidene)tricyclo[6.2.1.0²,⁷]undec-2(7)-en-3-ylidene]hydroxylamine

EC Number:

700-700-2

Cas Number:

1369492-55-6

Molecular formula:

C12H13Cl2NO

IUPAC Name:

N-[(3E)-11-(dichloromethylidene)tricyclo[6.2.1.0²,⁷]undec-2(7)-en-3-ylidene]hydroxylamine

Details on test material:

- Physical state: solid
- Stability under test conditions: not reported
- Storage condition of test material: At room temperature at about 20 °C

Test animals

Species:

other: EpiDerm tissues (three-dimensional human skin model)

Details on test animals or test system and environmental conditions:

No test animals were used.
EpiDerm tissues were supplied by MatTek Corporation, Ashland, Massachusetts, USA and stored refrigerated. Tissues were transferred to 6-well plates with the assay medium 90 minutes before starting the assay.

Test system

Type of coverage:

other: Not applicable

Preparation of test site:

other: Not applicable

Vehicle:

other: Not applicable - no vehicle used

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 40 mg
- Concentration (if solution): Undiluted test material

Duration of treatment / exposure:

Contact time of 3 minutes or 1 hour

Observation period:

Incubated at 37°C for 3 hours

Number of animals:

No animals were used on this test.
Four EpiDerm tissues were used per test substance, negative and positive control

Details on study design:

APPLICATION OF TEST AND CONTROL MATERIALS:
Immediately prior to treatment was intitiated the assay medium was replaced.
Approximately 40 mg of solid test substance was placed onto the tissue and moistened with 25 µL water to ensure good contact with the tissue surface. Further tissues were treated with 40 mL distilled water (negative control) and with 40 mL 8N potassium hydroxide (positive control).
Duplicate tissues were used for each contact time (3 minutes or 1 hour).
After the 3 minute or 1 hour contact periods, the tissues were washed with phosphate buffered saline (PBS) to remove residual material.

CELL VIABILITY MEASUREMENTS:
The assay medium was replaced with 300 mL of 1 mg/mL MTT-medium and tissues were incubated for 3 hours at 37°C. After incubation, the tissues were washed with PBS and formazan was extracted with 2 mL isopropanol overnight, without shaking.
The optical density of the extracted formazan was determined spectrophotometrically at 570 nm and cell viability was calculated for each tissue as a percentage of the mean of the negative control tissue. Skin corrosivity potential of the test article was classified according to the remaining cell viability obtained after test article treatment with either of the two treatment times.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Value:

105

Remarks on result:

other: Basis: mean

Remarks:

Time point: 3 minutes. Reversibility not measured.

Results 2

Irritation / corrosion parameter:

% tissue viability

Value:

101

Remarks on result:

other: Basis: mean.

Remarks:

Time point: 1 hour. Reversibility not measured.

Any other information on results incl. tables

Skin corrosion assay; cell viability measurements

3 minute treatment

Substance	Tissue replicate	OD540			Mean	Tissue mean	% difference between Rep’s	% Relative survival
		Aliquot 1	Aliquot 2	Aliquot 3
Negative control	A	1.521	1.527	1.527	1.525	1.617	10.8	100
	B	1.710	1.709	1.708	1.709
Test article	A	1.662	1.777	1.793	1.744	1.702	-5.0	105
	B	1.602	1.688	1.692	1.660
Positive control	A	0.418	0.428	0.426	0.424	0.425	-0.4	26
	B	0.424	0.425	0.428	0.426

        1 hour treatment

Substance	Tissue replicate	OD540			Mean	Tissue mean	% difference between Rep’s	% Relative survival
		Aliquot 1	Aliquot 2	Aliquot 3
Negative control	A	1.854	1.852	1.844	1.850	1.771	-9.3	100
	B	1.711	1.684	1.683	1.693
Test article	A	1.762	1.767	1.752	1.760	1.791	3.4	101
	B	1.823	1.841	1.804	1.823
Positive control	A	0.224	0.238	0.228	0.230	0.239	-8.0	13
	B	0.247	0.251	0.246	0.248

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance was considered not to be corrosive to the invitro skin testing model, EpiDerm.

Executive summary:

The result indicates that the substance should be non-corrosive to skin. Without furtherin vivodata the result has to be considered as inconclusive regarding classification.