Iron tris(2-ethylhexanoate)

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 18 to July 26, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

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:

June 2018

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Iron tris(2-ethylhexanoate)

IUPAC Name:

Iron tris(2-ethylhexanoate)

Test animals / tissue source

Details on test animals or tissues and environmental conditions:

- Justification of the test method and considerations regarding applicability : this method was selected as, based on the existing information, the substance was expected not to cause sufficient eye irritation to require a classification.
- Description of the cell system used, incl. certificate of authenticity and the mycoplasma status of the cell live: EpiOcularTM tissues

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 51.7 mg in Tissue 1 and 52.2 mg in Tissue 2.
- Concentration (if solution): applied undiluted

Duration of treatment / exposure:

6 hours

Duration of post- treatment incubation (in vitro):

18 hours

Number of animals or in vitro replicates:

2 replicates

Details on study design:

- Details of the test procedure used
- RhCE tissue construct used, including batch number : from MatTek In Vitro Life Science Laboratories, Mylnské Nivy 73, 82105 Bratislava, Slovakia. Batch n. 27046
- Doses of test chemical and control substances used : approx 52 mg of test substance undiluted, 50 µL of demineralised water as positive control and 50 µL of methyl acetate as negative control
- 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 nm
- Demonstration of proficiency in performing the test method before routine use by testing of the proficiency chemicals : the validity of the EpiOcularTM test was demonstrated in a proficiency study. For this purpose 15 proficiency chemicals (indicated by the OECD 492 guideline) were tested. All of the 15 proficiency chemicals were correctly categorized. Therefore, the proficiency of the EpiOcularTM test was demonstrated.

Results and discussion

In vitro

Results

Irritation parameter:

other: % Viability

Value:

77.8

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

other: Not eye irritant

Remarks:

according to the criteria set up on the OECD guideline 492

Conclusions:

Not eye irritant

Executive summary:

Method

The eye irritation potential of the test substance has been investigated according to the OECD guideline 492.

As the test item showed intense coloring in the pre-test, there was the risk to influence the photometric measurement. Therefore an additional test for intensely coloured test items was performed.

The viscous non-pipettable test item was treated as solid and therefore it was applied topically to a three-dimensional RhCE tissue construct in duplicate for an exposure time of 6 hours.

 

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.

Results

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 2.0. The positive control showed clear eye irritating effects, the mean value of the relative tissue viability was 38.2 % (< 50%).

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

 

After treatment with the test item, the mean value of relative tissue viability was 77.8 %.

Conclusion

Test items that induce values above the threshold are considered non-eye irritant, therefore, as the mean value of relative tissue viability found under this test conditions is above the threshold for eye irritation potential (≤ 60%), the test substance is considered non- eye irritant in the EpiOcular^TMEye Irritation Test.