Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05 December 2017 to 14 December 2017
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
Type:
Constituent
Test material form:
other: paste

Test animals / tissue source

Species:
human
Details on test animals or tissues and environmental conditions:
- Tissue :Reconstructed human Cornea-like Epithelium (RhCE)
- Model used: ‘EpiOcularTM irritation test ( MatTek Corporation EpiOcularTM EIT (OCL-200) Reconstituted Human Ocular Epithelium )
- Tissue batch number(s): (Lot # 27017)
- Delivery date: 12 December 2017
- Date of initiation of testing: no data

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 30 minutes ± 2 minutes, followed by a 12 ± 2 minutes’ post-treatment immersion at 37°C, 5% CO2
- Temperature of post-treatment incubation: 2 hours ± 15 minutes’ post-treatment incubation at 37°C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: not indicated (according to MatTEek SOP)

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE: no data (according to MatTek SOP)
- MTT concentration: not specified
- Incubation time: no data
- Spectrophotometer: not identified measured at 570 nm

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes
yes, concurrent positive control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µl
Duration of treatment / exposure:
30 minutes ± 2 minutes, followed by a 12 ± 2 minutes’ post-treatment immersion at 37°C, 5% CO2
Number of animals or in vitro replicates:
3 replicates
Details on study design:
tisues were wetted with 20 uL PBS before application of the substance or controls

negative control: sterile H2O
positive control: methyl acetate (neat)

number of replicates: 3

Results and discussion

In vitro

Results
Irritation parameter:
other: viability (%)
Value:
100.1
Negative controls validity:
valid
Remarks:
OD570 1.992
Positive controls validity:
valid
Remarks:
viability 40.03%
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
Acceptance criteria passed

Acceptance criterion 1 The mean OD570 of the negative control(treated with PBS) tissues is > 0.8 and < 2.8
 
Acceptance criterion 2 The mean of the positive control relative percentage viability must be below 50% of the mean of the negative controls.
 
Acceptance criterion 3 The standard deviationof OD values between three replicates in each experimental condition must be ≤ 18%
 

Any other information on results incl. tables

Name

Code

mean of OD

SD of OD

Mean of viability (%)

SD of viability (%)

CV %

Classification

Sterile water

NC

1.992

0.094

100.000

4.74

4.738

Non-Irritant

Methyl Acetate

PC

0.797

0.139

40.030

6.966

17.401

Irritant

Substance

TA1

1.994

0.049

100.100

2.467

2.465

No Category

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not irritant when tested in the EpiOcular TM Eye Irritation Test
Executive summary:

The substance was tested for eye irritation in the EpiOcular TM  Eye Irritation model according to OECD 492. 50 uL of the substance, positive control (methyl acetate) and negative control (H2O) was applied to the tissue for 30 minutes, followed by a 12 minutes post-treatment immersion at 37°C, 5% CO2 and thereafter incubated for 2 hour post-dose at 37°C, 5% CO2. 

The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 100.1% of negative controls, while validity criteria were met. The substance is concluded to be non-irritant.