Ethanaminium, N-[4-[[4-(diethylamino)phenyl][4-(ethylamino)-1-naphthalenyl]methylene]-2,5-cyclohexadien-1-ylidene]-N-ethyl-, molybdatetungstatephosphate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 August - 28 August 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Bis[[4-[[4-(diethylamino)phenyl][4-(ethylamino)-1-naphthyl]methylene]cyclohexa-2,5-dien-1-ylidene]diethylammonium] dicopper(1+) hexa(cyano-C)ferrate(4-)

EC Number:

279-935-3

EC Name:

Bis[[4-[[4-(diethylamino)phenyl][4-(ethylamino)-1-naphthyl]methylene]cyclohexa-2,5-dien-1-ylidene]diethylammonium] dicopper(1+) hexa(cyano-C)ferrate(4-)

Cas Number:

82338-76-9

Molecular formula:

C33H40N3.1/2C6FeN6.Cu

IUPAC Name:

bis[[4-[[4-(diethylamino)phenyl][4-(ethylamino)-1-naphthyl]methylene]cyclohexa-2,5-dien-1-ylidene]diethylammonium] dicopper(1+) hexa(cyano-C)ferrate(4-)

Test material form:

solid: nanoform

Remarks:

Purple powder

Details on test material:

specific surface area >100m2/cm3

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch no 278PE160900 from Client
- Expiration date of the lot/batch: 28 April 2022
- Purity test date: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Roomtemperature in the dark
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: Used as supplied
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: N/a

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None
- Preliminary purification step (if any): N/a
- Final dilution of a dissolved solid, stock liquid or gel: N/a
- Final preparation of a solid: Used as supplied

FORM AS APPLIED IN THE TEST (if different from that of starting material) Used as supplied

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: SkinEthic Laboratories

Source strain:

other: Reconstructed human epidermis

Amount/concentration applied:

Used as supplied = 100% (10 mg)

Duration of treatment / exposure:

15 mins

Number of replicates:

3

Test system

Controls:

yes

Details on study design:

An assessment of the test item’s capability to directly reduce MTT was inconclusive due to the intrinsic color of the test item. Therefore, an additional procedure using water killed tissues was performed. The results of the water killed tissues were subtracted from the mean OD of the test item treated viable tissues to obtain the true amount of MTT reduction that reflects metabolic conversion only.
The solution containing the test item was a blue color, therefore additional color correction tissues were incorporated into the testing procedure. The results of the color correction tissues were subtracted from the mean OD of the test item treated viable tissues to obtain the true amount of MTT reduction that reflects metabolic conversion only.
The results of the third set of control tissues were used to adjust the calculated correction factors derived from the color interference and MTT direct reduction controls in order to prevent a double correction from a colored test item that also reduces MTT. Mean OD570 values and correction factor calculations are given in the report.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean of three runs

Value:

44.7

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

The relative mean tissue viability for the positive control treated tissues was 10.7% relative to the negative control treated tissues and the standard deviation value of the viability was 2.8%. The positive control acceptance criteria were therefore satisfied.

The mean OD₅₇₀for the negative control treated tissues was 0.847 and the standard deviation value of the viability was 9.8%. The negative control acceptance criteria were therefore satisfied.

The standard deviation calculated from individual tissue viabilities of the three identically test item treated tissues was 11.3%. The test item acceptance criterion was therefore satisfied.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The test item was classified as irritant. The following classification criteria apply:
EU CLP and UN GHS Hazard statement H315 “Causes Skin Irritation” Category 2