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Administrative data

Description of key information

Key studies are available for both skin and eye irritation endpoints. All studies are performed in accordance with current OCED guidelines and under the conditions of GLP. As such the data is considered to be adequate and reliable for use as a key study and also for classification and labelling (Klimisch reliability 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:
17 October 2017 - 20 October 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 449935-248
- Expiration date of the lot/batch: 30 March 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: IN refrigerator (2-8ºC)
- Stability under test conditions: Stable under test conditions

Test system:
human skin model
Remarks:
EpiDerm Skin Model (EPI-200, Lot no.: 27161 Kit J and Kit K)
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Cells were purchased or derived from tissue obtained by MatTek Corporation from acredited institutions.
Source strain:
other: Keratinocyte strain: 4F1188
Justification for test system used:
Recommended test system in international guidelines (OECD and EC).
Vehicle:
unchanged (no vehicle)
Details on test system:
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1ºC (actual range 36.7 - 37.4°C).
- Temperature of post-treatment incubation (if applicable): 37ºC
REMOVAL OF TEST MATERIAL AND CONTROLS
- 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. The skin inserts were carefully dried.
DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Wavelength: 570 nm
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 4 per test item / 2 per control
PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the relevant mean tissue viability obtained after 3-minute tretment compared to the negative control tissues is decreased by below 50%. In addition, a test item considered nn-corosive (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.
- The test substance is considered to be non-corrosive to skin if the relative mean tissue viability obtained after the 3-minute treatment compared to the negative control tissues is not decreased below 50%. In addition, the relative tissue viabilty 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: 50μl

NEGATIVE CONTROL
- Amount(s) applied: 50µl

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50µl
- Concentration (if solution): 8N
Duration of treatment / exposure:
3 minutes / 1 hour
Number of replicates:
2 per exposure time
For the negative and positive controls, 2 tissues
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3-minute application
Value:
101
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1-hour application
Value:
90
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
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

RESULTS

2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate 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 mean absorption at 570 nm measured after treatment with the test item and controls are presented inAppendix 1, Table1. The individual OD570 measurements are presented in table 1.

Table 2 shows the mean tissue viability obtained after 3-minute and 1-hour treatments with the test item compared to the negative control tissues. Skin corrosion is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after the 3-minute and 1-hour treatments with the test item compared to the negative control tissues was 101% and 90% respectively. Because the mean relative tissue viability for the test item was not below 50% after 3 minutes treatment and not below 15% after 1 hour treatment the test item is considered to be not corrosive.

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 upper acceptance limit<=2.8) and the laboratory historical control data range. The mean relative tissue viability following the 1-hour exposure to the positive control was 7.7%. 

In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was < 8%, indicating that the test system functioned properly.

Table 1

Mean Absorption in the in vitro skin corrosion test with 2,2,2 -trifluoro-1 -(trifluoromethyl)ethyl methacrylate

         3 -minute application (Mean)           1 -hour application (Mean)
   A (OD570) B (OD570) Mean (OD570) SD  A (OD570)  B (OD570)  Mean (OD570) SD    
 Negative control  2.085 1.936   2.010±0.106 2.183  2.371  2.277± 0.133    
 Test Item  2.063 1.988  2.025± 0.053  2.063  2.045  2.054± 0.013    
 Positive control  0.131 0.143  0.137± 0.009  0.187  0.162  0.175± 0.018    

SD = Standard deviation

Duplicate exposures are indicated by A and B.

In this table the values are correct for background absorption (0.0430). Isopropanol was used to measure the background absorption.

Table 2

Mean tissue viability in the in vitro skin corrosion test with 2,2,2 -trifluoro-1 -(trifluoromethyl)ethyl methacrylate

   3 -minute application viability (percentage of control) 1 -hour application viability (percentage of control) 
Negative control  100  100 
 Test Item 101  90 
 Positive control 6.8  7.7 

Table 3

Coefficient of variation between tissue replicates

   3 -minute  1 -hour
 Negative control 7.2  7.9 
 Test item 3.6  0.9 
 Positive control 8.7  13 
Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, 2,2,2 -trifluoro-1-(trifluoromethyl)ethyl methacrylate is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this report.
Executive summary:

The objective of this study was to evaluate 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate for its ability to induce skin corrosion onon a human three dimensional epidermal model (EpiDerm (EPI-200)). The possible corrosive potential of the test item 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 449935-248 of the test item was a clear colourless liquid. The test item was applied undiluted (50 µl) directly on top of the skin tissue. 

The positive control had a mean relative tissue viability of 7.7% after the 1-hour exposure. 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. In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was < 8%, 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 the test item compared to the negative control tissues was 101% and 90%, respectively. Because the mean relative tissue viability for the test item was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment the test item is considered to be not corrosive.

In conclusion, 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate 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:
November to December 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 449935-248
- Expiration date of the lot/batch: 30 March 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In refrigerator (2-8ºC)
- Stability under test conditions: Stable
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimize the need of in vivo testing. One of the validated in vitro skin irritation tests is the EPISKIN test, which is recommended in international guidelines (e.g. OECD and EC).
Vehicle:
unchanged (no vehicle)
Details on test system:
SKIN DISC PREPARATION
- Procedure used: On the day of receipt the tissues were transferred to 12-well plates and pre-incubated with pre-warmed Maintenance Medium for 24 hours at 37°C. Maintenance medium and Assay medium were supplied by Skinethic Laboratories, Lyon, France.
- Quality control for skin discs: Electrical resistance obtained with two of the isolated skin discs was [complete, e.g. 10 kΩ]

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
- Number of washing steps: 2

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: TECAN Infinite M200 Pro Plate Reader
- Wavelength: 570

NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if [complete, e.g. if the mean TER value is less than or equal to 5 kΩ and the skin disk is obviously damaged, or if the mean TER value is less than or equal to 5 kΩ, and the skin disc is showing no obvious damage, but the mean disc dye content is greater than or equal to the mean disc dye content of the 10M HCl positive control obtained concurrently.]
- The test substance is considered to be non-corrosive to skin if [complete, e.g. if the mean TER value obtained for the test substance is greater than 5 kΩ, or if the mean TER value is less than or equal to 5 kΩ, and the skin disc is showing no obvious damage, and the mean disc dye content is well below the mean disc dye content of the 10M HCl positive control obtained concurrently.]
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25µl

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 25µl

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 25µl
- Concentration (if solution): 5%
Duration of treatment / exposure:
15 ± 0.5 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Mean Tissue Viability
Value:
94
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
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

Acceptability Criteria

The in vitro skin irritation test is considered acceptable if it meets the following criteria:

a)     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 and the Standard Deviation value (SD) of the % viability should be <=18.

b)     The mean relative tissue viability of the positive control should be <= 50% relative to the negative control and the Standard Deviation value (SD) of the % viability should be <= 8.

c)     The SD calculated from individual % tissue viabilities of the three identically treated replicates should be <=8.

All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented.

Table 1: Data interpretation of test item

 Relative mean viability of 3 individual tissues after 15 minutes of exposure and 42 hours of post incubation Prediction to be considered 
 <= 50% of the menu viability of the negative controls Categry 2 
 > 50% of the mean viability of the negative controls No category 

Calculation of Cell Viability

Optical Density readings were transferred into Microsoft Excel to allow further calculations to be performed.

The corrected OD (ODc) for each sample or control was calculated by subtracting the value of the blank mean (ODbl) from each reading (ODraw).

ODc= ODraw– ODbl

The OD value representing 100% cell viability is the average OD of the negative controls (ODlt_u+MTT). 

The % Viability for each sample and positive control is calculated as follows:

%Viability = (ODc/mean ODlt_u+MTT) * 100

Results

2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate was checked for possible direct MTT reduction and color interference in the Skin corrosion test using EpiDerm as a skin model. Because no color changes were observed it was concluded that the test item did not interact with the MTT endpoint. 

The mean absorption at 570 nm measured after treatment with the test item and controls are presented in Table 2.

The individual OD570 measurements are presented in.

Table 3 shows the mean tissue viability obtained after 15 ± 0.5 minutes treatment with the test item compared to the negative control tissues. 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 the test item compared to the negative control tissues was 94%. Since the mean relative tissue viability for the test item was above 50% the test item is considered to be non-irritant.

The positive control had a mean cell viability after 15 ± 0.5 minutes exposure of 7.0%. The absolute mean OD570 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 < 9%, indicating that the test system functioned properly.

Table 2: Mean Absorption in the In Vitro Skin Irritation Test with the test item

  A (OD570  B (OD570)  C (OD570)  Mean (OD570)  SD
 Negative control 1.090  0.962  0.991  1.015  +- 0.067 
 The test item 0.906  1.053  0.915  0.958 +-  0.082 
 Positive control 0.052 0.100  0.060  0.071 +-  0.025 

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 (0.042). Isopropanol was used to measure the background absorption.

Table 3: Mean Tissue Viability in the In Vitro Skin Irritation Test with the teset item

  Mean tissue viability (percentage of control)  Standard deviation (percentage) 
 Negative control 100  6.6 
 The test item 94  8.1 
 Positive control 7.0  2.5 

Table 4: Individual OD Measurements at 570nm

   A (OD570)  B (OD570)  C (OD570)

 Negative control

OD570 measurement 1

OD570 measurement 2

1.1153

1.1501 

 

1.0068

1.0022

1.0755

0.9913 

 The test item

OD570 measurement 1

OD570 measurement 2

 

0.9587

0.9378

1.1100

1.0807

0.9678

0.9476 

 Positive control

OD570 measurement 1

OD570 measurement 2

 

0.0950

0.0946

0.1383

0.1457 

0.1073

0.0973 

                              

OD = Optical density

Triplicate exposures are indicated by A, B and C.

Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report and should not be classified.
Executive summary:

The objective of this study was to evaluate 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate for its ability to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small model (EPISKIN-SMTM)). The possible skin irritation potential of the test item was tested through topical application for 15 minutes.

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

Batch 449935-248 of the test item was a clear colourless liquid. The test item was applied undiluted (25 µl), directly on top of the skin tissue for 15 ± 0.5 minutes. 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 the test item compared to the negative control tissues was 94%. Since the mean relative tissue viability for the test item was above 50% after 15 ± 0.5 minutes treatment the test item is considered to be non-irritant.

The positive control had a mean cell viability of 7.0% after 15 ± 0.5 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 < 9%, indicating that the test system functioned properly.

In conclusion, 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report and should not be classified.

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:
October - November 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
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)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 449935-248
- Expiration date of the lot/batch: 30 March 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In refridgerator (2-8ºC)
Species:
other: Bovine eyes
Strain:
other: Bovine eyes
Details on test animals or tissues and environmental conditions:
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.
Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.
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): 750µl
- Concentration (if solution): as supplied
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
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 ± 1ºC. The corneas were incubated for the minimum of 1 hour at 32 ± 1ºC.

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.
QUALITY CHECK OF THE ISOLATED CORNEAS

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
A negative control, physiological saline (Eurovet Animal Health, Bladel, The Netherlands) was included to detect non-specific changes in the test system and to provide a baseline for the assay endpoints.

POSITIVE CONTROL USED
Ethanol, Identification number RS532, Batch number K47177483

APPLICATION DOSE AND EXPOSURE TIME
750µl of either the negative control, positive control (Ethanol) or test item. 10 minute exposure time

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 2
- POST-EXPOSURE INCUBATION: 120 ± 10 minutes

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=(I_0/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: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value)
Additionally the opacity and permeability values were evaluated independently to determine whether the test item induced irritation through only one of the two endpoints.

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

DECISION 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.
All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented.
Irritation parameter:
cornea opacity score
Run / experiment:
mean opacity score
Value:
-0.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
in vitro irritation score
Run / experiment:
mean in vitro irritation score
Value:
-0.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes

Results

2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate was tested neat. 

Table1 summarizes the opacity, permeability andin vitroirritancy scores of the test item 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 0.3 to 2.3. The individual positive control in vitro irritancy scores ranged from 37 to 53. The corneas treated with the positive control item were turbid after the 10 minutes of treatment.

The corneas treated with the test item showed opacity values ranging from -0.8 to -0.4 and permeability values ranging from 0.001 to 0.003. The corneas were clear after the 10 minutes of treatment with the test item. No pH effect of the test item was observed on the rinsing medium. Hence, the in vitro irritancy scores ranged from -0.8 to -0.3 after 10 minutes of treatment with 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate.

Table 1: Summary of Opacity, Permeability and In Vitro Scores

 Treatment Mean Opacity1   Mean Permeability1  Mean In Vitro Irritation Score1,2
 Negative control 1.6  -0.004  1.5 
 Positive control (ethanol)  21 1.751  48 
 Test Item -0.5  0.002  -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).

Table 2: Opacity Scores

 Treatment Opacity before treatment  Opacity after treatment  Final Opacity1   Negative control corrected Final Opacity2  Mean Final Opacity
 Negative control 1.0  1.5  0.4    1.6 
   3.2 5.1  2.0     
   4.9 7.3  2.4     
 Positive control 1.4  21.4  20.1  18  21 
   1.7 26.5  24.8  23   
   1.7 26.1  24.4  23   
 Test Item 2.4  3.5  1.1  -0.5  -0.5 
   4.3 5.1  0.8  -0.8   
   2.7 3.9  1.2  -0.4   

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

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

3    Calculations are made without rounding off.

Table 3: Permeability Score individual Values (Uncorrected)

 Treatment Dilution factor  OD490 1  OD490 2  OD490 3  Average OD  Final OD  Mean final negative control 
 Negative control -0.010  -0.010  -0.008  -0.009  -0.009  -0.004 
   1 -0.003  -0.004  -0.004  -0.001  -0.001   
   1 -0.003  -0.003  -0.002  -0.003  -0.003   
 Positive control 1.219  1.235  1.234  1.229  1.229   
   6 0.325  0.331  0.333  0.0330  1.978   
   6 0.326  0.327  0.342  0.332  1.990   
 Test Item -0.005  -0.006  0.001  -0.003  -0.003   
   1 -0.001  -0.002  -0.004  -0.002 

-0.002 

 
   1 -0.002  -0.003  0.002  -0.001  -0.001   

Table 4: Permeability Score Individual Values (corrected)

 Treatment Dilution factor  Negative control corrected OD490 11    Negative control corrected OD490 21   Negative control corrected OD490 31  Negative control corrected OD490 Average Negative control corrected final OD490  Average OD 
 Positive control 1.223  1.239  1.238  1.234  1.234  1.751 
   6 0.329  0.335  0.337  0.334  2.004   
   6 0.330  0.331  0.346  0.336  2.016   
 Test Item -0.001  -0.002  0.005  0.001  0.001  0.002 
   1 0.003  0.002  0.000  0.002  0.002   
   1 0.002  0.001  0.006  0.003  0.003   

1    OD490 values corrected for the mean final negative control permeability (-0.004).

2    Calculations are made without rounding off.

Table 5: In Vitro Irritancy Score

Treatment Final Opacity  Final OD4902  In vitro Irritancy Score1
 Negative control 0.4  -0.009  0.3 
   2.0 -0.001  1.9 
   2.4 -0.003  2.3 
 Positive control 18  1.234  37 
   23 2.004  53 
   23 2.016  53 
 Test Item -0.5  0.001  -0.5 
   -0.8 0.002  -0.8 
   -0.4 0.003  -0.3 

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

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

Interpretation of results:
GHS criteria not met
Conclusions:
Since 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate 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 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate 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 the test item was tested through topical application for 10 minutes. 

The study procedures described in this report were based on the most recent OECD guideline.

Batch 449935-248 of the test item was a clear colourless liquid. 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 limits of the laboratory historical rangeindicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 48 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. 

The test item 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 2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

The data available for skin and eye irritation of 2,2,2 -trifluoro-1-(trifluoromethyl)ethyl methacrylate concludes that no classification is required according to Regulation (EC) No. 1272/2008 (EU CLP).