3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 Feb - 10 Mar 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

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted Jul 2015

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

EC Number:

246-791-8

EC Name:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

Cas Number:

25291-17-2

Molecular formula:

C8H3F13

IUPAC Name:

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooct-1-ene

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: LabCyte EPI-MODEL24 SIT

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: LabCyte EPI-MODEL24 SIT (Japan Tissue Engineering Co., Ltd.)
- Tissue batch number(s): LCE24-170206-A
- Delivery date: 07 Feb 2017
- Expiry date: 10 Feb 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Tissues were rinsed fifteen times or more with PBS (without Mg2+ and Ca2+). Remaining PBS was removed from outside of tissue insert with a sterile cotton swab.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.5 mg/mL
- Incubation time: 180 ± 5 min
- Spectrophotometer: Multimode Microplate Reader (FLUOstar OPTIMA, BMG LABTECH)
- Wavelength: 570 and 650 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: The quality of the LabCyte EPI-MODEL 24 tissue was assessed by undertaking a MTT cell viability test. The OD was determined to be 1.1 (acceptance criteria 0.8 ≤ OD ≤ 2.5).
- Barrier function: According to the Supplier`s Data Sheet, the IC50 value was determined to be 0.26 (acceptance criteria: 0.14 ≤ IC50 ≤ 0.40).
- Morphology: Observing multilayered epidermis with a stratum corneum according to the Supplier`s Data Sheet.

NUMBER OF REPLICATE TISSUES: Triplicate tissues

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
The test substance did not directly reduce MTT.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability is less than or equal to 50%.
- The test substance is considered to be non-corrosive to skin if the viability is greater than 50%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 25 µL

NEGATIVE CONTROL
- Amount(s) applied: 25 µL

POSITIVE CONTROL
- Amount(s) applied: 25 µL
- Concentration: 5% (w/v)

Duration of treatment / exposure:

15 min ± 30 s (administered in 1 min intervals)

Duration of post-treatment incubation (if applicable):

42 ± 1 h

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

15 min ± 30 s

Value:

118

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction: The test substance did not show a MTT-reducing capacity.
- Tissue-binding test: As a result of tissue-binding test, the staining ratio was -0.2%, thus below 5% and a correction of measurement value was not conducted.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD measured after treatment with the negative control was 0.810, thus in the acceptability range (0.8 ≤ OD ≤ 2.5).
- Acceptance criteria met for positive control: The cell viability was strongly reduced in tissues treated with the positive control. The mean cell viability was 2.7%.
- Acceptance criteria met for variability between replicate measurements: The standard deviations of the cell viabilities in the negative control, positive control and test substance group were 10.8, 0.1 and 1.8%, respectively, and within the acceptability range for the variability (SD ≤ 18.0) according to OECD TG 439.

Any other information on results incl. tables

Table 1: Summary of Results

Group	Tissue Number	OD	Mean OD	Cell Viability (%)	Mean Cell Viability (%)	SD
Negative control	1	0.902	0.810	111.4	100	10.8
	2	0.801		98.9
	3	0.727		89.8
Positive control	1	0.022	0.022	2.7	2.7	0.1
	2	0.022		2.7
	3	0.021		2.6
Test substance	1	0.960	0.956	118.5	118.0	1.8
	2	0.940		116.0
	3	0.967		119.4

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation(EC) No. 1272/2008

Conclusions:

Under the conditions of the RHE test method the test substance did not show irritant properties.
CLP: not classified