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

Eye irritation

Administrative data

Endpoint:

eye 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 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

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

Test animals / tissue source

Species:

human

Strain:

other: Epiocular™EIT (OCL-200)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 50 µL

NEGATIVE CONTROL
- Amount(s) applied: 50 µL
- Lot/batch no.: K6G73

POSITIVE CONTROL
- Amount(s) applied: 50 µL
- Lot/batch no.: 020917ALA

Duration of treatment / exposure:

30 ± 2 min (administere in 1 min intervals)

Duration of post- treatment incubation (in vitro):

12 ± 2 min

Number of animals or in vitro replicates:

duplicates for each treatment and control group; from each tissue, 2 absorbance measurements after MTT incubation were performed

Details on study design:

- RhCE tissue construct used, including batch number: Epiocular™EIT (OCL-200); Test Kit name: OCL-200EIT (manufactured on 16 Feb 2017 by MatTek Corporation); Lot number: 20978; Receipt Date: 20 Feb 2017

- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods:

- Indication of controls used for direct MTT-reducers and/or colouring test chemicals: The ability of the test substance to directly reduce MTT and to change the color of the MTT medium was assessed in a pre-experiment. Since the MTT solution colour did not change, the test substance is not presumed to have reduced the MTT. A tissue-binding test was performed with medium without MTT to determine the staining ratio using the following formula:
Staining ratio (%) = OD of the test substance (without MTT) / Mean OD of the negative control substance group (with MTT) x 100

- Number of tissue replicates used per test chemical and controls: 2

- Wavelength: 570 nm

- Spectrophotometer: Multimode Microplate Reader (FLUOstar OPTIMA, BMG Labtech)

- Description of evaluation criteria used: The test substance is considered to be not irritating to eye if the tissue viability is > 60%. The test substance is considered to be irritating to eye if the tissue viability is ≤ 60%.

- Positive and negative control means and acceptance ranges: OD in the negative control substance group is > 0.8 and < 2.5. The cell viability in the positive control substance group is < 50%.
- Acceptable variability between tissue replicates for positive and negative controls and for the test substance: Differences of two tissue cell viabilities in each treatment group are < 20%

Results and discussion

In vitro

Results

Irritation parameter:

other: % tissue viability mean value of 2 tissues

Run / experiment:

30 min exposure

Value:

72.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
The optical pre-experiment (colour interference pre-experiment) to investigate the colour change potential of the test substance in MTT medium did not led to a change in colour.
The mean staining ratio was below 60%, therefore the cell viability was corrected by the following formula:
Corrected cell viability (%) = Mean cell viability of the test substance group (%) - Mean staining ratio (%)

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes, the OD of the negative control was 1.117, 1.052, 1.005 and 1.047, and thus within the acceptibility range (OD > 0.8 and < 2.5).
- Acceptance criteria met for positive control: Yes, the positive control reduced the cell viability at 36.0% and fulfilled the acceptance criteria.
- Acceptance criteria for variabilties: Differences of two tissue cell viabilities in the negative control substance, the positive control substance and the test substance groups were 5.5%, 4.2% and 2.4%, respectively, and thus < 20%.

Any other information on results incl. tables

Table 1: Summary of Results

Group	Tissue	OD	Mean OD / Tissue	Mean OD of Tissue 1 and 2	Mean Cell Viability (%) / Tissue	Mean Cell Viability (%) of Tissue 1 and 2	Corrected Mean Cell Viability (%) *	Difference
Negative control	1	1.117	1.085	1.056	102.7	100	-	5.5
		1.052
	2	1.005	1.026		97.2
		1.047
Test substance	1	0.394	0.402	0.380	38.1	36.0	-	4.2
		0.410
	2	0.351	0.358		33.9
		0.364
Positive control	1	0.775	0.786	0.773	74.4	73.2	72.4	2.4
		0.797
	2	0.745	0.760		72.0
		0.774

* The cell viability was corrected by the staining ratio

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 RhCE test method the test substance did not show irritant properties.
CLP: not classified