Tetradecafluorohexane

Eye irritation

Administrative data

Endpoint:

eye irritation: 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

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

adopted October 09, 2017

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

Specific details on test material used for the study:

Test item information
Identification: Tétradécafluorohexane
Appearance: Clear colourless liquid
Batch: 57 A
Purity/Composition: 99.25%
Test item storage: At room temperature protected from light
Stable under storage conditions until: 21 February 2020 (expiry date)
Additional information
Test Facility test item number: 209384/A
Purity/Composition correction factor: No correction factor required
Test item handling: No specific handling conditions required
Stability at higher temperatures: Stable

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands),
where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter.
Transport: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 %µL

Duration of treatment / exposure:

10 +/- 1 minutes

Duration of post- treatment incubation (in vitro):

120 +/- 10 minutes

Number of animals or in vitro replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Value:

ca. -0.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

1.6

Positive controls validity:

valid

Remarks:

40

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Value:

ca. -0.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

1.8

Positive controls validity:

valid

Remarks:

16

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

fluorescein retention score

Value:

ca. 0.008

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

-0.0016

Positive controls validity:

valid

Remarks:

1.625

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. Altough the negative control treated corneas were translucent, all results are within the acceptability range therefore this has no impact on the study result. The mean in vitro irritancy score of the positive control (Ethanol) was 40 and within two standard deviations of the current historical positive control mean (Appendix 3, Table 6). It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Any other information on results incl. tables

Tétradécafluorohexane was tested neat.

Table 1of Appendix 1summarizes the opacity, permeability and in vitro irritancy scores of Tétradécafluorohexane and the controls. The opacity, permeability and in vitro scores of the individual corneas are shown in Table 2- 5.

The individual in vitro irritancy scores for the negative controls ranged from 0.8 to 2.7. The corneas treated with the negative control item were translucent after the 10 minutes of treatment. All values were within the historical control database. The individual positive control in vitro irritancy scores ranged from 31 to 48 (Appendix 2,Table 5). The corneas treated with the positive control item were turbid after the 10 minutes of treatment.

The corneas treated with Tétradécafluorohexane showed opacity values ranging from -0.9 to

-0.5 and permeability values ranging from 0.006 to 0.011. The corneas were translucent after the 10 minutes of treatment with Tétradécafluorohexane. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -0.7 to -0.4 after 10 minutes of treatment with Tétradécafluorohexane.

Table 1

Summary of Opacity, Permeability and In Vitro Scores

Treatment	Mean Opacity1	Mean Permeability1	Mean In vitro Irritation Score1, 2
Negative control	1.8	-0.016	1.6
Positive control (Ethanol)	16	1.625	40
Test item	-0.7	0.008	-0.5

1        Calculated using the negative control mean opacity and mean permeability values for the positive control and test item.

2        In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

Individual Opacity, Permeability and in vitro Score

Table 2 Opacity Score

Treatment	Opacity before treatment	Opacity after treatment	Final Opacity1		Negative control corrected Final Opacity2	Mean Final Opacity
Negative control	4.7	6.2	1.4			1.8
	4.5	5.5	1.0
	3.9	6.8	2.9
Positive control	2.0	18.3	16.3	14		16
	3.5	20.8	17.4	16
	4.4	23.3	18.9	17
Test item	3.7	4.8	1.2	-0.6		-0.7
	3.9	5.2	1.3	-0.5
	4.4	5.3	0.9	-0.9

Calculations are made without rounding off.

1Final Opacity = Opacity after treatment – Opacity before treatment.

2Negative control corrected Final Opacity = Final opacity – Mean final opacity negative control.

Table 3

Permeability Score Individual Values (Uncorrected)

 

Treatment	Dilutionfactor	OD490 1	OD490 2	OD490 3	Average OD	Final OD	Mean final negative control
Negative control	1	-0.017	-0.017	-0.017	-0.017	-0.017	-0.016
	1	-0.016	-0.016	-0.016	-0.016	-0.016
	1	-0.016	-0.016	-0.016	-0.016	-0.016
Positive control	6	0.272	0.276	0.277	0.275	1.650
	6	0.349	0.345	0.342	0.345	2.072
	1	0.931	0.943	0.945	0.940	0.940
Test item	1	-0.010	-0.010	-0.011	-0.010	-0.010
	1	-0.009	-0.010	-0.011	-0.010	-0.010
	1	-0.010	-0.008	0,001	-0.009	-0.009

Calculations are made without rounding off.

Table 4

Permeability Score Individual Values (Corrected)

 

Treatment	Dilution factor	Negative control correctedOD4901^1	Negative control correctedOD4902^1	Negative control correctedOD4903^1	Negative control correctedOD490 Average	Negative control corrected final OD490	Average OD
Positive control	6	0.288	0.292	0.293	0.291	1.748	1.625
	6	0.365	0.361	0.358	0.362	2.170
	1	0.947	0.959	0.961	0.956	0.956
Test item	1	0.006	0.006	0.005	0.006	0.006	0.008
	1	0.007	0.006	0.005	0.006	0.006
	1	0.006	0.008	0.017	0.011	0.011

Calculations are made without rounding off.

1OD490values corrected for the mean final negative control permeability (-0.016).

 

Table 5

In Vitro Irritancy Score

 

Treatment	Final Opacity2	Final OD  2 490	In vitro Irritancy Score1
Negative control	1.4	-0.017	1.2
	1.0	-0.016	0.8
	2.9	-0.016	2.7
Positive control	14	1.748	41
	16	2.170	48
	17	0.956	31
Test item	-0.6	0.006	-0.5
	-0.5	0.006	-0.4
	-0.9	0.011	-0.7

1    In vitro irritancy score (IVIS) = opacity value + (15 x OD490value).

2    Positive control and test item are corrected for the negativecontrol.

Table 6

Historical Control Data for the BCOP Studies

 

	Negative control			Positive control
	Opacity	Permeability	In vitro Irritancy Score	In vitro Irritancy Score
Range	-2.9 – 3.0	-0.034 – 0.100	-2.8 – 3.0	28.0 – 110.9
Mean	0.25	0.00	0.31	55.03
SD	1.13	0.01	1.19	15.08
n	118	118	118	94

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 May 2015 to May 2018.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, since Tétradécafluorohexane induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

Executive summary:

The objective of this study was to evaluate the eye hazard potential of Tétradécafluorohexane as measured by its ability to induce opacity and increase permeability in an isolated bovine cornea using the Bovine Corneal Opacity and Permeability test (BCOP test).

This report describes the potency of chemicals to induce serious eye damage using isolated bovine corneas. The eye damage of Tétradécafluorohexane was tested through topical application for 10 minutes.

The study procedures described in this report were based on the most recent OECD guideline. Batch 57 A of Tétradécafluorohexane was a clear colorless liquid with a purity of 99.25%.

The test item was applied as it is (750 µL) directly on top of the corneas.

The negative control responses for opacity and permeability were less than the upper limitsof the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 40 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Tétradécafluorohexane did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of -0.5 after 10 minutes of treatment.

In conclusion, since Tétradécafluorohexane induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.