Tetradecafluorohexane

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tetradecafluorohexane

EC Number:

206-585-0

EC Name:

Tetradecafluorohexane

Cas Number:

355-42-0

Molecular formula:

C6F14

IUPAC Name:

tetradecafluorohexane

Test material form:

liquid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

Test system
EpiDerm Skin Model (EPI-200, Lot no.: 28356, kit A, Appendix 4).
The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis.

It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.
Rationale
Recommended test system in international guidelines (OECD and EC). Source
MatTek Corporation, Ashland MA, U.S.A.

Amount/concentration applied:

50 µL

Duration of treatment / exposure:

3 minutes & 1 hour

Number of replicates:

2 x 2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

3 minutes

Value:

ca. 97

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100

Positive controls validity:

valid

Remarks:

11

Remarks on result:

other: No indication of corrosion

Results 2

Irritation / corrosion parameter:

% tissue viability

Value:

ca. 95

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100

Positive controls validity:

valid

Remarks:

5.6

Remarks on result:

other: No indication of corrosion

Other effects / acceptance of results:

Tétradécafluorohexane was checked for color interference in aqueous conditions and possible direct MTT reduction by adding the test item to MTT medium. Because the solutions did not turn blue / purple nor a blue / purple precipitate was observed it was concluded that the test item did not interfere with the MTT endpoint.
The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance limit <2.8) and the laboratory historical control data range (See Appendix 3). The mean relative tissue viability following the 1-hour exposure to the positive control was 5.6%.
In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was
< 17%, indicating that the test system functioned properly (Appendix 1, Table 3).

Any other information on results incl. tables

Table 1

Mean Absorption in the in vitro Skin Corrosion Test with Tétradécafluorohexane

 

	A(OD570)	3-minute application B(OD570)         Mean (OD570)			SD	1-hour application A (OD570)  B(OD570)                                    Mean (OD570)				SD
Negative control	1.609	1.929	1.769	±	0.227	1.838	1.843	1.841	±	0.003
Tétradécafluorohexane	1.774	1.656	1.715	±	0.084	1.831	1.650	1.741	±	0.129
Positive control	0.247	0.131	0.189	±	0.082	0.089	0.115	0.102	±	0.018

SD = Standard deviation

Duplicate exposures are indicated by A and B.

In this table the values are corrected for background absorption (0.0423). Isopropanol was used to measure the background absorption.

Table 2

Mean Tissue Viability in the in vitro Skin Corrosion Test with Tétradécafluorohexane

 

	3-minute application viability (percentage of control)	1-hour application viability (percentage of control)
Negative control	100	100
Tétradécafluorohexane	97	95
Positive control	11	5.6

 

Table 3

Coefficient of Variation between Tissue Replicates

 

	3 minute	1 hour
Negative control	17	0.3
Tétradécafluorohexane	6.7	9.9
Positive control	47	22

CV (%) = 100 - [(lowest OD570/highest OD570) x 100%]

Individual OD Measurements at 570 nm

	3-minute application(OD570) A              B		1-hour application(OD570) A              B
Negative control OD570 measurement 1	1.6578	1.9712	1.8766	1.9152
OD570 measurement 2	1.6490	1.9674	1.8973	1.8840
OD570 measurement 3	1.6464	1.9758	1.8675	1.8570
Test itemOD570 measurement1	1.8177	1.6722	1.9150	1.7002
OD570 measurement 2	1.8213	1.6912	1.8495	1.6773
OD570 measurement 3	1.8107	1.7306	1.8568	1.6985
Positive control OD570 measurement 1	0.2894	0.1751	0.1334	0.1601
OD570 measurement 2	0.2863	0.1716	0.1327	0.1559
OD570 measurement 3	0.2915	0.1722	0.1288	0.1567

OD = Optical density

Duplicate exposures are indicated by A and B.

Historical Control Data for in vitro Skin Corrosion Studies

	Negative control		Positive control
	3-minutetreatment (OD570)	1-hourtreatment (OD570)	3-minutetreatment (OD570)	1-hourtreatment (OD570)
Range	1.258 – 2.615	1.371 – 2.371	0.0172 – 0.56	0.046 – 0.339
Mean	1.80	1.82	0.19	0.14
SD	0.26	0.22	0.09	0.05
n	111	110	106	103

SD = Standard deviation

n = Number of observations

The above mentioned historical control data range of the controls were obtained by collecting all data over the period of November 2014 to November 2017.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Tétradécafluorohexane is not corrosive in the in vitro skin corrosion test under the experimental conditions described in the report.

Executive summary:

The objective of this study was to evaluate Tétradécafluorohexane for its ability to induce skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential of Tétradécafluorohexane was tested through topical application for 3 minutes and 1 hour.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch 57 A of Tétradécafluorohexane was a clear colourless liquid. Tétradécafluorohexane was applied undiluted (50 µL) directly on top of the skin tissue.

The positive control had a mean relative tissue viability of 5.6% after the 1-hour exposure.The absolute mean OD570(optical density at 570 nm) of the negative control tissues waswithin the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upperacceptance limit£2.8) and the laboratory historical control data range. In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was£17%, indicating that the test system functioned properly.

Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with Tétradécafluorohexane compared to the negative control tissues was 97% and 95%, respectively. Because the mean relative tissue viability for Tétradécafluorohexane was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment Tétradécafluorohexane is considered to be not corrosive.

In conclusion, Tétradécafluorohexane is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.