Registration Dossier

Diss Factsheets

Toxicological information

Eye irritation

Currently viewing:

Administrative data

eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report

Materials and methods

Test guideline
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)
Version / remarks:
The additional non-specific controls with additional freeze-killed tissues without MTT addition were not performed, since in the performed additional tests with freeze-killed tissues and with viable tissues without MTT addition the gained absorption values were very low and did not have a detrimental impact on the outcome of the study. An additional test with freeze-killed tissues without MTT addition would have resulted in a very slight increase of the test item viability value in the decimal place, and therefore, also would not have changed the result relevantly.
There was the following deviation from the study plan without an influence on the outcome and the validity of the study: see remarks.
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2-(1,3-benzodioxol-5-ylamino)ethanol hydrochloride
EC Number:
EC Name:
2-(1,3-benzodioxol-5-ylamino)ethanol hydrochloride
Cas Number:
Molecular formula:
2-(1,3-benzodioxol-5-ylamino)ethanol hydrochloride
Test material form:
solid: crystalline

Test animals / tissue source

Details on test animals or tissues and environmental conditions:
human-derived epidermal keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea. It consists of highly organized basal cells which progressively flatten out as the apical surface of the tissue is approached, analogous to the normal in vivo corneal epithelium.

Test system

yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
50 mg
Duration of treatment / exposure:
6 hours
Duration of post- treatment incubation (in vitro):
18 hours
Number of animals or in vitro replicates:

Results and discussion

In vitro

Irritation parameter:
other: cell viability [%]
Negative controls validity:
Positive controls validity:
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
The acceptance criteria were met.

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
In conclusion, it can be stated that in this study and under the experimental conditions reported, Hydroxyethyl-3,4-methylenedioxyaniline HCL possesses an eye irritating potential.
Executive summary:

This in vitro study was performed to assess the eye irritation potential of Hydroxyethyl-3,4-methylenedioxyaniline HCL by means of the Human Cornea Model Test.

The test item proved to be an MTT reducer in the MTT pre-test. Also it proved to dye water and isopropanol in the colour interference pre-test. Therefore, additional tests with freeze-killed and with viable tissues had to be performed. The viability values resulted in these additional tests were used to correct the values gained in the main experiment.

Each 50 mg of the test item, were applied to each of duplicate tissue for 6 hours. Each 50 µL of the negative control (deionised water) and of the positive control (methyl acetate) were also applied to duplicate tissues each. After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD > 0.8 and < 2.5 thus showing the quality of the tissues. Treatment with the positive control induced a decrease below 50% compared with thenegative control value in the relative absorbance thus ensuring the validity of the test system. The difference of viability between the two relating tissues was < 20% in the same run (for test item tissues, positive and negative control tissues of the main experiment). Irritating effects were observed following incubation with Hydroxyethyl-3,4-methylenedioxyaniline HCL. Compared with the negative control the relative mean absorption value corresponding to the viability of the tissues decreased below 60% (6.5%).

In conclusion, it can be stated that in this study and under the experimental conditions reported, Hydroxyethyl-3,4-methylenedioxyaniline HCL possesses an eye irritating potential.