[No public or meaningful name is available]

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:

6.5.-5.7.2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2019

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,1,3,3-tetrachloroprop-1-ene

EC Number:

675-808-5

Cas Number:

18611-43-3

Molecular formula:

C3H2Cl4

IUPAC Name:

1,1,3,3-tetrachloroprop-1-ene

Test material form:

liquid

Details on test material:

concentration 99,87%

In vitro test system

Test system:

human skin model

Cell type:

non-transformed keratinocytes

Cell source:

other: EpiDerm (Source: MatTek Corporation, USA) is a three-dimensional human epidermis model.

Vehicle:

not specified

Details on test system:

3-minute exposure: After the 1-hour pre-incubation, each tissue was treated with an exposure
time of 3 minutes at room temperature.
60-minute exposure: After the 1-hour pre-incubation, the “Maintenance Medium” was replaced
with fresh medium (0.9 mL per well). Each tissue was treated with an exposure time of
60 minutes. The tissues were placed at 37 °C in an incubator with 5% CO2, in a ≥95%
humidified atmosphere for the exposure time.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 μL of the test item was applied evenly to the epidermal surface with a pipette.

Duration of treatment / exposure:

3-minute exposure and 60minute exposure

Duration of post-treatment incubation (if applicable):

15 min

Number of replicates:

two for each test ( 3 min and 60 min§

Test system

Type of coverage:

not specified

Vehicle:

other: MTT diluent (MTT-100-DIL)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50 μL of the test item was added to 1 mL MTT working solution

Duration of treatment / exposure:

After the 1-hour pre-incubation, each tissue was treated with an exposure
time of 3 minutes at room temperature

Observation period:

Pre-warmed (37 °C) MTT medium was added to each well below the skin units. The lid was
replaced and the plate was incubated at 37°C in an incubator with 5 % CO2, in a ≥95%
humidified atmosphere for 3 hours (± 15 minutes).
The MTT medium was removed from each well; the tissues were rinsed with DPBS (filling and
removing the wells three times), then dried and placed into a new plate.
2 ml extractant (isopropanol) was added per well, which covered inside and outside the tissue
insert. The plates were sealed shaken for two hours on a plate shaker at room temperature for
extraction.
After the extraction, the tissues were pierced, letting the extractant inside the insert to flow into
the well. The plates were shaken at approx. 120 rpm for 15 minutes.

Details on study design:

Following the formazan extraction, 3 × 200 μL sample from each tube was placed into the wells
of a 96-well plate (labelled appropriately). The OD (Absorbance / Optical Density) of the
samples was read in a 96-well plate spectrophotometer (FLUOstar Optima microplate reader,
BMG LABTECH GmbH), at a wavelength of 570 ± 10 nm, using the extractant (isopropanol) as
blank (6 × 200 μL). Linearity range of measuring device was determined as up to 2.804 during
its validation.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2 tissues 3 min, 60 min, negative and positive control

Value:

ca. 35

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

Following exposure with 1133-Tetrachloropropene, the mean cell viability was 98.5% after the
3-minute exposure, and 34.9% after the 60-minute exposure, compared to the concurrent
negative control. This is above the thresholds of 50% (3-min) and more than 15% (60-min). As
the mean relative viability is above 50% after 3 minutes and 15% after 60 minutes of exposure,
the test item is considered not to be corrosive to skin.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, in this in vitro EpiDerm™ model test with 1133-Tetrachloropropene, the
results indicate that the test item is not corrosive to the skin.

Executive summary:

In conclusion, in this in vitro EpiDerm™ model test with 1133-Tetrachloropropene, the
results indicate that the test item is not corrosive to the skin.