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Toxicological information

Skin irritation / corrosion

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Administrative data

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

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 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate
EC Number:
221-309-9
EC Name:
2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate
Cas Number:
3063-94-3
Molecular formula:
C7H6F6O2
IUPAC Name:
2,2,2-trifluoro-1-(trifluoromethyl)ethyl methacrylate
Test material form:
liquid
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

In vitro test system

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

Results and discussion

In vitro

Resultsopen allclose all
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

Any other information on results incl. tables

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 

Applicant's summary and conclusion

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.