Registration Dossier

Diss Factsheets

Toxicological information

Eye irritation

Currently viewing:

Some information in this page has been claimed confidential.

Administrative data

eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018 January 25 - 2018 May 31
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guidelineopen allclose all
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)
according to guideline
other: EURL ECVAM DB-ALM Method Summary No. 164: EpiOcular™ Eye Irritation Test
according to guideline
other: EpiOcular™ Eye Irritation Test (OCL-200-EIT)
Version / remarks:
For the prediction of acute ocular irritation of chemicals For use with MatTek Corporation’s Reconstructed Human EpiOcular™ Model
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Test material form:
Details on test material:
Clear oily liquid, molecular weight 462.61, relative density 1.025
Specific details on test material used for the study:
- Source and lot/batch No.of test material: Vertellus, 0000140180
- Expiration date of the lot/batch: 15 June 2018

- Storage condition of test material: room temperature
- Stability under test conditions: assumed to be stable

Test animals / tissue source

Details on test animals or tissues and environmental conditions:
The test was carried out with the EpiOcular™ reconstructed human cornea-line epithelium (RhCE) model (MatTek). The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a stratified, highly differentiated squamous epithelium morphologically similar to that found in a human cornea. The EpiOcular™ RhCE tissue construct consists of at least 3 viable layers of cells and a non-keratinised surface, showing a cornea-like structure analogous to that found in vivo.

Test system

unchanged (no vehicle)
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
50 mg
Duration of treatment / exposure:
6 hr
Duration of post- treatment incubation (in vitro):
18 hr
Number of animals or in vitro replicates:
Details on study design:
- Details of the test procedure used : RhCE tissue was treated with test substance for 6 hrs. After an 18 hr post-treatment incubation, tissue samples were evaluated for viability using an MTT reduction assay.
- RhCE tissue construct used, including batch number : EpiOcular™ tissues on agarose (Lot No.: 27019 for NSMTT control, 27039 for main experiment)
- Doses of test chemical and control substances used : Negative control: 50 µL. Positive control: 50 µL. Test item: 50 mg. The test item was ground into a fine powder using a mortar and pestle. Approximately 50 mg (83.3 mg/cm^2) of the test item were applied directly to the EpiOcular TM tissue using an application spoon, avoiding compression of the test item. The test item was spread to match size of the tissue.
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods (where applicable) : Exposure: 6 hr, 37 °C. Post-exposure immersion: 25 min, room temperature. Post-exposure incubation: 18 hr, 37 °C.
- Justification for the use of a different negative control than ultrapure H2O (if applicable) : N/A.
- Justification for the use of a different positive control than neat methyl acetate (if applicable) : N/A
- Description of any modifications to the test procedure : N/A
- Indication of controls used for direct MTT-reducers and/or colouring test chemicals (if applicable) : The mixture of 50 mg test item per 1 mL MTT medium showed reduction of MTT as compared to the solvent. The mixture turned blue/purple. Since the mean relative tissue viability of the test item treated tissues (TM) was above the 60% threshold value killed tissue controls were performed for quantitative correction of results.
- Number of tissue replicates used per test chemical and controls (positive control, negative control, NSMTT, NSCliving and NSCkilled, if applicable) : 2
- Wavelength and band pass (if applicable) used for quantifying MTT formazan, and linearity range of measuring device (e.g. spectrophotometer) : 570
- Description of the method used to quantify MTT formazan : Absorbance (OD570)
- Description of the qualification of the HPLC/UPLC-spectrophotometry system (if applicable) : N/A
- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model : Ocular irritation potential of the test item was predicted from the relative mean tissue viabilities obtained after treatment compared to the negative control tissues concurrently treated with Aqua dest. (distilled water). The test item is considered to be irritant to the eye but it cannot be differentiated between UN GHS “Category 1” or “Category 2”, if the relative tissue viability is less or equal to 60%. The test item is considered to be non-irritant in accordance with UN GHS “No Category” if relative tissue viability is higher than 60%.
- Reference to historical positive and negative control results demonstrating suitable run acceptance criteria : Yes
- Complete supporting information for the specific RhCE tissue construct used : Yes
- Reference to historical data of the RhCE tissue construct: Yes
- Demonstration of proficiency in performing the test method before routine use by testing of the proficiency chemicals : Yes
- Positive and negative control means and acceptance ranges based on historical data : Yes
- Acceptable variability between tissue replicates for positive and negative controls: Yes
- Acceptable variability between tissue replicates for the test chemical: Yes

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
other: Tissue Viability [%]
Run / experiment:
Negative Control
Vehicle controls validity:
not applicable
Negative controls validity:
not applicable
Positive controls validity:
Remarks on result:
no indication of irritation
Irritation parameter:
other: Tissue Viability [%]
Run / experiment:
Positive Control
Vehicle controls validity:
Negative controls validity:
not applicable
Positive controls validity:
not applicable
Remarks on result:
positive indication of irritation
Irritation parameter:
other: Tissue Viability [%]
NSMTT corrected
Run / experiment:
Test item
Vehicle controls validity:
Negative controls validity:
not applicable
Positive controls validity:
Remarks on result:
no indication of irritation
Other effects / 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: Observed values in range of historical values

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
The test substance was evaluated for eye irritation potential according to OECD Guideline 492. In this study under the given conditions the test item showed no irritant effects. The test item is classified as “non-irritant“ in accordance with UN GHS “No Category”.