Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Skin corrosion / irritation: not corrosive (OECD 431, GLP, K, rel.1), not irritant (OECD 439, GLP, K, rel.1)

Eye irritation/damage: not classified (OECD 437, GLP, K, rel. 1)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
25-Oct-2016 to 26-Jan-2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted according to OECD test Guideline No. 431 and EU Method B.40 BIS. Furthermore, functional model conditions and references to historical control data are included in the report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
Adopted July 29, 2016
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: UE Method B.40 BIS (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
Following the REACH bottom-up strategy, the EpiDerm™ Human Skin Model method was used to assess skin corrosion as recommended in the OECD test guideline No. 431.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Epi-200-Kit, MatTek Corporation (Ashland, MA, USA).
- Lot number: 24940 Kits J and K
- Production Date: no data
- Shipping date: no data
- Delivery date: no data
- Date received: no data
- Date of initiation of testing: December 06, 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 and 60 minutes at 37 °C in a humidified atmosphere of 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: After the exposure period, the tissues were washed with phosphate buffered saline (Invitrogen Corporation, Breda, The Netherlands) to remove residual test item. Rinsed tissues were kept in 24 well plates on 300 μl DMEM medium until 6 tissues (= one application time) were dosed and rinsed.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL MTT solution
- Incubation time: 3 hours at 37 °C, 5% CO2 in air
- Spectrophotometer: TECAN Infinite® M200 Pro Plate Reader
- Wavelength: 570 nm
- Filter: without reference filter
- Linear OD range of spectrophotometer: not reported

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: OD (540-570 nm): 1.744 +/- 0.093 [1.0-3.0]
- Barrier function: ET-50: 5.21 hrs [4.77-8.72 hrs]
- Morphology: 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.
- Contamination: No contamination

NUMBER OF REPLICATE TISSUES: 4 tissues per test item together with a negative control and positive control

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Not needed

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
A test item is considered corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after 3-minute treatment compared to the negative control tissues is decreased below 50%.
b) In addition, a test item considered non-corrosive (viability >= 50%) after the 3-minute treatment is considered corrosive if the relative tissue viability after 1-hour treatment with the test item is decreased below 15%.
A test item is considered non corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after the 3-minute treatment compared to the negative control tissues is not decreased below 50%.
b) In addition, the relative tissue viability after the 1-hour treatment is not decreased below 15%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL of the undiluted test item.

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of Milli-Q water was used as supplied

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of 8.0 N Potassium Hydroxide was used as supplied
Duration of treatment / exposure:
3 and 60 minutes.
Duration of post-treatment incubation (if applicable):
not applicable
Number of replicates:
4 tissues per test item together with a negative control and positive control
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minutes exposure
Value:
98
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
7%
Remarks on result:
other: No indication of corrosion
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
60 minutes exposure
Value:
46
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
14%
Remarks on result:
other: No indication of corrosion
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: Not reported
- Direct-MTT reduction: None
- Colour interference with MTT: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes

Table 7.3.1/1: Mean absorption in the in vitro skin corrosion test with Perfluoro methoxy dioxole

     3 -minute application              1 -hour application
 A (OD570)  B (OD570)

 Mean (OD570)

   SD  A (OD570)  B (OD570)  Mean (OD570)    SD
 Negative control 1.698 1.618  1.658   +/- 0.056  1.440  1.371  1.406   +/- 0.049 
 Test item 1.719 1.531  1.625   +/- 0.133  0.678  0.608  0.643   +/- 0.049 
 Positive control 0.125 0.114  0.120   +/- 0.008  0.234  0.170  0.202   +/- 0.046 

OD = Optical Density

SD = Standard Deviation

Duplicate exposures are indicated by A and B.

In this table the values are corrected for background absorption (0.0424). Isopropanol was used to measure the background absorption.

Table 7.3.1/2: Mean tissue viability in the in vitro skin corrosion test with Perfluoro methoxy dioxole

 

 3-minute application viability

(percentage of control)

 1-hour application viability

(percentage of control)

 Negative control  100 (4.7)  100 (4.8)
 Test item  98 (11) 46 (10) 
 Positive control  7 (9.0)  14 (28)

( ): Coefficient of variation between tissue replicates

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, Perfluoro methoxy dioxole is not corrosive to skin.
Executive summary:

An in vitro skin corrosion study was performed according to the most recent OECD Guideline 431, EU Method B.40 BIS and in compliance with GLP, using the EpiDerm™ Human Skin Model.

 

The test item was applied undiluted (50 μl for 3 minutes exposure and an excess amount for the 1-hour exposure) directly on top of the skin tissue. Since the test item was volatile, an excess amount of the test item was applied every 15 minutes for the 1-hour exposure.

The positive control had a mean relative tissue viability of 14% after the 1-hour exposure. This is within the acceptability range for the positive control (<15%). 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 (1.324 – 2.615 for 3 minute exposure and 1.361 – 2.352 for 1 hour exposure). In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was =< 11%, indicating that the test system functioned properly (acceptability value:≤30%).

The relative mean tissue viability obtained after 3-minute and 1-hour treatments with the test item compared to the negative control tissues was 98% and 46%, respectively. Because the mean relative tissue viability for Perfluoro methoxy dioxole was not below 50% compared to control after the 3-minute treatment and not below 15% after the 1-hour treatment Perfluoro methoxy dioxole is considered to be not corrosive.

Finally, it is concluded that this test is valid and that Perfluoro methoxy dioxole is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17-Jan-2017 to 21-Mar-2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted according to OECD test Guideline No. 439 and EU Method B.46. Furthermore, functional model conditions and references to historical control data are included in the report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
adopted 28 July 2015
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
23 July 2009
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Adult donors
Justification for test system used:
Following the REACH top-down strategy, the EPISKIN™ Reconstructed Human Epidermis Model method was used to assess skin irritation as recommended in the OECD test guideline No. 439.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN™ Reconstructed Human Epidermis Model Kit, SkinEthic Laboratories, Lyon, France
- Tissue batch number(s): 17-EKIN-008
- Production date: not reported
- Shipping date: not reported
- Delivery date: not reported
- Expiry date: 27 February 2017
- Date of initiation of testing: 17 January 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation (if applicable): 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: After the exposure period of 15 ± 0.5 minutes at room temperature, the tissues were washed with phosphate buffered saline to remove residual test item. After rinsing, the cell culture inserts were each dried carefully and moved to a new well on 2 ml pre-warmed maintenance medium until all tissues were dosed and rinsed. Subsequently the skin tissues were incubated for 42 hours at 37°C. Number of washing steps not reported.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none reported

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: TECAN Infinite® M200 Pro Plate Reader
- Wavelength: 570 nm
- Filter: without reference filter
- Linear OD range of spectrophotometer: not reported

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Barrier function: IC50 = 1.8 mg/mL [1.5-3.0 mg/mL]
- Morphology: Well-differenciated epidermis consisting of a basal layer, several spinous and granular layers and a thinck stratum corneum
- Contamination: absence of bacteria, fungus and mycoplasma

NUMBER OF REPLICATE TISSUES: 3 tissues per test item together with negative and positive controls

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE:
Not needed

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
A test item is considered irritant in the skin irritation test if:
The relative mean tissue viability of three individual tissues after 15 minutes of exposure to the test item and 42 hours of post incubation is ≤ 50% of the mean viability of the negative controls.
A test item is considered non-irritant in the in vitro skin irritation test if:
The relative mean tissue viability of three individual tissues after 15 minutes of exposure to the test item and 42 hours of post incubation is > 50% of the mean viability of the negative controls.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- The test item was used as supplied (undiluted).
- Amount(s) applied (volume or weight with unit): 25 μL directly on top of the tissue. Due to the volatility of the test item, the tissues were re-treated three times with the undiluted test item.

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 25 μL
- Concentration (if solution): Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 25 μL
- Concentration (if solution): 5% (aq) Sodium dodecyl sulphate
Duration of treatment / exposure:
15 +/- 0.5 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3 tissues per test item together with negative and positive controls
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15 minute exposure period and 42 h post-exposure incubation period
Value:
73
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
22%
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: Not reported
- Direct-MTT reduction: None
- Colour interference with MTT: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: Not included in the report

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes
- Range of historical values if different from the ones specified in the test guideline:
The absolute mean OD570 (optical density at 570 nm) of the three tissues of the negative control should reasonably be within the laboratory historical control data range (0.676 - 1.336) (assays performed from November 2013 to November 2016).

Table 7.3.1/3: Mean absorption in the in vitro skin irritation test with Perfluoro methoxy dioxole

   A (OD570)  B (OD570)  C (OD570)  Mean (OD570)    SD
 Negative control  0.826 0.872   0.808  0.835  +/-  0.033
 Test item  0.648  0.540  0.648  0.612  +/-  0.062
 Positive control  0.193  0.224  0.139  0.186  +/-  0.043

OD = Optical Density

SD = Standard Deviation

Triplicate exposures are indicated by A, B and C.

In this table the values are corrected for background absorption (0.041). Isopropanol was used to measure the background absorption.

Table 7.3.1/4: Mean tissue viability in the in vitro skin irritation test with Perfluoro methoxy dioxole

 

Mean tissue viability (percentage of control)

 Negative control  100 (3.9)
 Test item  73 (7.5)
 Positive control  22 (5.2)

( ): Standard deviation (percentage)

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, Perfluoro methoxy dioxole is non-irritant to skin.
Executive summary:

An in vitro skin irritation study was performed according to the OECD Guideline 439, EU Method B.46 and in compliance with GLP, using the EPISKINTM reconstructed human epidermis model.

Perfluoro methoxy dioxole was applied undiluted (25 μl) directly on top of the skin tissue for 15 ± 0.5 minutes. Due to the volatility of the test item, the tissues were re-treated three times with the undiluted test item. After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Skin irritation is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with Perfluoro methoxy dioxole compared to the negative control tissues was 73%. Since the mean relative tissue viability for Perfluoro methoxy dioxole was above 50% after 15 ± 0.5 minutes treatment Perfluoro methoxy dioxole is considered to be non-irritant.

The positive control had a mean cell viability of 22% after 15 minutes exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 8%, indicating that the test system functioned properly.

Finally, it is concluded that this test is valid and that Perfluoro methoxy dioxole is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07-Apr-2017 to 01-Sep-2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted according to OECD test Guideline No. 437. Furthermore, functional model conditions and references to historical control data are included in the report.
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 26 July 2013
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Assessed on 07-11, 14 and 16 September 2015. Dated on the 03 November 2015.
Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco,’s Hertogenbosch, The Netherlands)
- Number of animals: Not specified
- Characteristics of donor animals (e.g. age, sex, weight): Not specified
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Eyes were collected and transported in physiological saline in a suitable container under cooled conditions
- Time interval prior to initiating testing: as soon as possible after slaughter
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): undiluted
Duration of treatment / exposure:
10 +/- 1 minutes at 32 +/- 1°C
Duration of post- treatment incubation (in vitro):
120 +/- 10 minutes at 32 +/- 1°C
Number of animals or in vitro replicates:
3 corneas were selected at random for each treatment group
Details on study design:
SELECTION AND PREPARATION OF CORNEAS
The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded.
The isolated corneas were stored in a petri dish with cMEM (Earle’s Minimum Essential Medium (Life Technologies, Bleiswijk, The Netherlands) containing 1% (v/v) L-glutamine (Life Technologies) and 1% (v/v) Foetal Bovine Serum (Life Technologies)). The isolated corneas were mounted in a corneal holder (one cornea per holder) of BASF (Ludwigshafen, Germany) with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was positioned on top of the cornea and tightened with screws. The compartments of the corneal holder were filled with cMEM of 32 +/- 1°C. The corneas were incubated for the minimum of 1 hour at 32 +/- 1°C.

QUALITY CHECK OF THE ISOLATED CORNEAS
After the incubation period, the medium was removed from both compartments and replaced with fresh cMEM. Opacity determinations were performed on each of the corneas using an opacitometer (BASF-OP3.0, BASF, Ludwigshafen, Germany). The opacity of each cornea was read against a cMEM filled chamber, and the initial opacity reading thus determined was recorded. Corneas that had an initial opacity reading higher than 7 were not used. Three corneas were selected at random for each treatment group.

NUMBER OF REPLICATES
3 corneas were selected at random for each treatment group

NEGATIVE CONTROL USED
physiological saline (Eurovet Animal Health, Bladel, The Netherlands)

POSITIVE CONTROL USED
Ethanol (Purity >= 99.9%)

APPLICATION DOSE AND EXPOSURE TIME
The medium from the anterior compartment was removed and 750 µL of either the negative control, positive control (Ethanol) or test item was introduced onto the epithelium of the cornea. The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the control or the test item over the entire cornea. Corneas were incubated in a horizontal position for 10 +/- 1 minutes at 32 +/- 1°C.

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD:
After the incubation the solutions were removed and the epithelium was washed with MEM with phenol red (Earle’s Minimum Essential Medium, Life Technologies) and thereafter with cMEM. Possible pH effects of the test item on the corneas were recorded. The medium in the posterior compartment was removed and both compartments were refilled with fresh cMEM. Subsequently the corneas were incubated for 120 +/- 10 minutes at 32 +/- 1°C. After the completion of the incubation period opacity determination was performed. Each cornea was inspected visually for dissimilar opacity patterns.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity:
The opacity of a cornea was measured by the diminution of light passing through the cornea. The light was measured as illuminance (I = luminous flux per area, unit: lux) by a light meter.
The opacity value (measured with the device OP-KIT) was calculated according to:

Opacity = [(I0/I) -0.9894] / 0.0251

With I0 the empirically determined illuminance through a cornea holder but with windows and medium, and I the measured illuminance through a holder with cornea.
The change in opacity for each individual cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final post-treatment reading. The corrected opacity for each treated cornea with the test item or positive control was calculated by subtracting the average change in opacity of the negative control corneas from the change in opacity of each test item or positive control treated cornea.
The mean opacity value of each treatment group was calculated by averaging the corrected opacity values of the treated corneas for each treatment group.

- Corneal permeability:
Following the final opacity measurement, permeability of the cornea to Na-fluorescein (Sigma-Aldrich, Germany) was evaluated.
The medium of both compartments (anterior compartment first) was removed. The posterior compartment was refilled with fresh cMEM. The anterior compartment was filled with 1 mL of 4 mg Na-fluorescein (Sigma-Aldrich Chemie GmbH, Germany)/ml cMEM solution. The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the sodium-fluorescein solution over the entire cornea. Corneas were incubated in a horizontal position for 90 +/- 5 minutes at 32 +/- 1°C.
After the incubation period, the medium in the posterior compartment of each holder was removed and placed into a sampling tube labelled according to holder number. 360 μl of the medium from each sampling tube was transferred to a 96-well plate. The optical density at 490 nm (OD490) of each sampling tube was measured in triplicate using a microplate reader (TECAN Infinite® M200 Pro Plate Reader). Any OD490 that was 1.500 or higher was diluted to bring the OD490 into the acceptable range (linearity up to OD490 of 1.500 was verified before the start of the experiment). OD490 values of less than 1.500 were used in the permeability calculation.
The mean OD490 for each treatment was calculated using cMEM corrected OD490 values. If a dilution has been performed, the OD490 of each reading of the positive control and the test item was corrected for the mean negative control OD490 before the dilution factor was applied to the reading.

- Others (e.g, pertinent visual observations, histopathology): Additionally the opacity and permeability values were evaluated independently to determine whether the test item induced irritation through only one of the two endpoints.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS) = mean opacity value + (15 x mean OD490 value)

DECISION CRITERIA:
The IVIS cut-off values for identifying the test items as inducing serious eye damage (UN GHS Category 1) and test items not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given hereafter:

In vitro score range: UN GHS:
=< 3 No Category
> 3 ; =< 55 No prediction can be made
> 55 Category 1

ACCEPTABILITY CRITERIA
The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.
Irritation parameter:
in vitro irritation score
Value:
-0.8
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: No, the corneas were clear after the 10 minutes of treatment with Perfluoro methoxy dioxole.

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Range of historical values if different from the ones specified in the test guideline:
The positive control should elicit an In Vitro Irritancy Score that falls within two standard deviations of the historical mean (2 x SD 12.64 to 56.68) for the laboratory.
The negative control mean opacity change value should be ≤3.0 and the permeability mean value ≤0.042 (assays performed from February 2014 to February 2017).
Other effects:
No pH effect of the test item was observed on the rinsing medium.

Table 7.3.2/1 - Summary of Opacity, Permeability and In Vitro Scores

 Treatment  Mean Opacity 1  Mean Permeability 1  Mean In Vitro Score 1, 2
 Negative control  -0.5  0.003  -0.4
 Positive control (Ethanol)  19.6  2.737  60.7
 Test item  -1.2  0.027  -0.8

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).

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, Perfluoro methoxy dioxole induced an IVIS =< 3, therefore, the test substance is not classified for eye irritation or serious eye damage, according to Regulation (EC) No. 1272/2008 (CLP) and to the UN GHS Regulation.
Executive summary:

The eye hazard potential of the Perfluoro methoxy dioxole was evaluated using the Bovine Corneal Opacity and Permeability test (BCOP test), under GLP compliance, according to the OECD Guideline No. 437.

The corneal damage potential of test substance was assessed using fresh bovine corneae. 750 µL of test item was applied to cornea for 10 minutes followed by an incubation period of 2 hours at 32 ± 1 °C and corneal opacity was measured. Three corneas were used for each treated series (undiluted test item; negative control: physiological saline; positive control: ethanol). Following the opacity measurement, the permeability of the corneas to sodium fluorescein was evaluated. Two endpoints, corneal opacity and permeability, were measured and combined to give an In Vitro Irritancy Score.

 

The individual in vitro irritancy scores for the negative controls ranged from -1.2 to 0.3. The individual positive control in vitro irritancy scores ranged from 47 to 72. The corneas treated with the positive control item were turbid after the 10 minutes of treatment. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range (3.0 and 0.042, respectively) indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 61 and within two standard deviations of the current historical positive control mean (34.7 - 78.2). It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The corneas treated with Perfluoro methoxy dioxole were clear after the 10 minutes of treatment with Perfluoro methoxy dioxole. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -2.1 to 0.7 after 10 minutes of treatment with Perfluoro methoxy dioxole and, the mean IVIS was lower than or equal to 3.

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion:

Two key studies have been identified.

A key study (CRL, 2017a, Rel.1) was performed to evaluate the corrosive potential of the test substance for skin. This in vitro skin corrosion study was performed according to the OECD Guideline 431 and in compliance with GLP, using the EpiDerm™ Human Skin Model. The relative mean viability of the test item treated tissues was 98.0% and 46.0% after 3 and 60 minutes exposure to the test item, respectively. The quality criteria required for acceptance of results in the test were satisfied. With a tissue viability > 50% after 3 minutes of exposure and > 15% after 60 minutes of exposure, the test material was considered not to be corrosive to skin.

A key study (CRL, 2017b, Rel.1) was performed to evaluate the irritant potential of the test substance for skin. This in vitro skin irritation study was performed according to the OECD Guideline 439 and in compliance with GLP, using the EPISKIN™ Reconstructed Human Epidermis Model. The relative mean viability of the test item treated tissues was 73.0% after 15 minute exposure to the test item followed by 42 hour post-treatment incubation. The quality criteria required for acceptance of results in the test were satisfied. With a tissue viability > 50% after exposure and post-treatment incubation, the test material was considered as non-irritant to skin.

Eye irritation:

A key study was identified for eye irritation. This BCOP assay (CRL, 2017c, Rel.1) was performed according to the OECD Guideline 437 and in compliance with GLP, using bovine corneas. The quality criteria required for acceptance of results in the test were satisfied. The mean In Vitro Irritancy Score of the test item was -0.8, after the 10 -minute exposure period followed by 120 -minute incubation period. With an IVIS =< 3, the test substance was considered as non-irritant to the eyes.

Justification for classification or non-classification

Harmonized classification:

The substance does not have an harmonized classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Based on the available data no additional self-classification is proposed for the substance regarding skin and eye irritation according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the UN GHS.

No data was available regarding respiratory irritation, however the substance not being classified for skin and eye irritation, it is unlikely that the substance requires to be classified as respiratory irritant.