2-({[4-({bis[4-({[2-(prop-2-enoyloxy)ethoxy]carbonyl}amino)phenoxy](sulfanylidene)-$l^{5}-phosphanyl}oxy)phenyl]carbamoyl}oxy)ethyl prop-2-enoate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

November 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-({[4-({bis[4-({[2-(prop-2-enoyloxy)ethoxy]carbonyl}amino)phenoxy](sulfanylidene)-λ⁵-phosphanyl}oxy)phenyl]carbamoyl}oxy)ethyl prop-2-enoate

EC Number:

918-919-9

Cas Number:

1072454-85-3

Molecular formula:

C36H36N3O15PS

IUPAC Name:

2-({[4-({bis[4-({[2-(prop-2-enoyloxy)ethoxy]carbonyl}amino)phenoxy](sulfanylidene)-λ⁵-phosphanyl}oxy)phenyl]carbamoyl}oxy)ethyl prop-2-enoate

additive 1

Reference substance name:

Ethyl acetate

EC Number:

205-500-4

EC Name:

Ethyl acetate

Cas Number:

141-78-6

Molecular formula:

C4H8O2

IUPAC Name:

ethyl ethanoate

Test material form:

liquid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

commercially available test system

Vehicle:

other: ethyl acetate

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: epiCS® (CellSystems, Troisdorf, Germany).
- Tissue batch number(s): Cat.-No.CS-1001
- Date of initiation of testing: 25 Nov 2015
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature for 3 min- exposure and incubator (37 ± 2 °C) for 60 min- exposure
- Temperature of post-treatment incubation (if applicable): 37 ± 2 °C
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: EL808, Bio-Tek
- Wavelength: 570 nm
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION:
the optical density of the isopropanol-extracts of 3 insert was determined by duplicate per insert = 6 OD values.
PREDICTION MODEL / DECISION CRITERIA:
- The mean optical density (OD) value obtained with the test item was used to calculate the percentage of viability relative to the negative control, which is set at 100 %.
- classification according to UN GHS: if the mean percent tissue viability after 3 min-exposure is less than (<) 50 % the test item is classified as corrosive (Category 1A); if the mean percent tissue viability after 3-min-exposure is greater than or equal (≥) to 50 % AND < 15 % after 60-min exposure the test item is classified as corrosive (Category 1B/1C); if the mean percent tissue viability after 60-min-exposure is ≥ 50 % after 3-min exposure AND ≥ 15 % after 60-min exposure the test item is considered as non-corrosive

The corrosive potential of the test item is assessed by determination of its cytotoxic effect on an in vitro reconstructed human epidermis. The test principle is based on the MTT assay reflecting the cell viability after exposure to the topically applied test item.
All tests were performed in triplets for each time point. The test item was applied at a 100 % concentration, i.e. 50 µL per insert for 3 min. (room temperature) and 60 min. Cell viability was measured by the amount of MTT reduction (calculated on the basis of optical density of the negative control).

Control samples:

yes, concurrent negative control

other: yes, solvent of the test material

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 50 μL
- the test item and the solvent ethyl acetate were tested in parallel

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 0.9 % NaCl

Duration of treatment / exposure:

3 min and 60 min

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

exposure period: 3 min. - result for test material

Value:

ca. 100

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks on result:

other: the solvent ethyl acetate was tested in parallel and showed 95 % tissue viability

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

exposure period: 60 min. - result for test material

Value:

ca. 61.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks on result:

other: the solvent ethyl acetate was tested in parallel and showed 19 % tissue viability

Other effects / acceptance of results:

DEMONSTRATION OF TECHNICAL PROFICIENCY:
- Reliability of the test was previously confirmed by inter-laboratory validation

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes, within regular intervals in the lab
- Acceptance criteria met for variability between replicate measurements: yes

Any other information on results incl. tables

Sample No.	Test item	Time [min.]	% Viability
1 -3	negative control NaCl 0.9 %	60	100.00
4 -6	test item	60	61.20
7 - 9	solvent ethyl acetate	60	18.52
16 -18	control NaCl 0.9 %	3	100.00
19 - 21	test item	3	99.63
22 - 24	solvent ethyl acetate	3	95.05

Applicant's summary and conclusion

Interpretation of results:

other: no corrosive property

Executive summary:

An in vitro study for predicting non-specific corrosive properties of the test item was conducted according to OECD TG 431. The undiluted test item (solution in ethyl acetate; 50 µL per insert) was applied topically to a reconstructed human epidermis model (RhE; epiCS). In parallel the solvent ethyl acetate was tested in this assay to distinguish potential toxicity of the registered substance from solvent toxicity. After an exposure period of 3 min at room temperature and 60 minutes the cell viability was 61.2 % and about 100 %, respectively, for the test item as measured by a MTT conversion assay. It is therefore concluded that the test item is not corrosive to the skin. Also the solvent ethyl acetate did not show corrosive properties according to the criteria of this assay.