Registration Dossier

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:
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)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Test material form:
liquid
Details on test material:
clear, colourless liquid
Production date: 14. Jan. 2018
Expiry date: 14. Jan. 2021
Storage: room temperature (20 ± 5 °C)
Batch no.: 701146_43343114

Test animals / tissue source

Details on test animals or tissues and environmental conditions:
EpiOcularTM tissues were procured from MatTek In Vitro Life Science Laboratories,
Mylnské Nivy 73, 82105 Bratislava, Slovakia.
Designation of the kit: OCL-200-EIT
Day of delivery: 24. Jul. 2018
Batch no.: 27060

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
50 μL of the liquid test item
Duration of treatment / exposure:
28 min
Duration of post- treatment incubation (in vitro):
115 min
Number of animals or in vitro replicates:
n=2

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
other: relative tissue viability
Run / experiment:
1
Value:
91.4
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
other: relative tissue viability
Run / experiment:
2
Value:
95.2
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
All validity criteria were fulfilled.
Values for negative control and for positive control were within the range of historical data of the test facility.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, the test item is considered non- eye irritant in the EpiOcularTM Eye Irritation Test.
Executive summary:

One valid experiment was performed.

The test item was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 28 minutes.

50 μL of the liquid test was applied to two tissue replicates.

After treatment, the respective substance was rinsed from the tissue; then, cell viability of the tissues was evaluated by addition of MTT, which can be reduced to formazan. The formazan production was evaluated by measuring the optical density (OD) of the resulting solution.

Demineralised water was used as negative control and methyl acetate was used as positive control. The controls showed the following results: After treatment with the negative control, the absorbance values were within the required acceptability criterion of mean OD > 0.8 and < 2.5, OD was 1.9. The positive control showed clear eye irritating effects, the mean value of the relative tissue viability was 34.7 % (< 50%).

Variation within tissue replicates of the controls and test item was acceptable (< 20%).

After treatment with the test item, the mean value of relative tissue viability was 93.3 %.

This value is well above the threshold for eye irritation potential (≤ 60%). Test items that induce values above the threshold are considered non-eye irritant.