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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27.02.2014
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Ocular Irritation Protocol: Neat Method (MTT ET-50) for use with EpiOcular TM Tissues, Rev: 17. Jan. 2012 by MatTek.
Deviations:
no
Principles of method if other than guideline:
SOP 118 00 890, edition 2 dated 01. Aug. 2013 ,“Bestimmung der hornhautreizenden Eigenschaften (Ermittlung ET50) mit EpiOcularTM“Additional: Stern M., Klausner M., Alvarado R., Renskers K., Dickens M., 1998. Evaluation of the EpiOcular Tissue Model as an Alternative to the Draize Eye Irritation Test. Toxicology in Vitro 12, 455-461
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
Name Tar wood Batch no. 18092013 Appearance dark brown, viscous liquid Composition Mixture CAS No. 91722-33-7 EINECS-No. not stated Molecular formula not stated Molecular weight not stated Purity 100% mixture Homogeneity not stated Vapour pressure not stated Stability not stated Solubility not indicated by the sponsor Production date not stated Expiry date 31. Dec. 2015 Storage Room temperature 20 ± 5 °C

Test animals / tissue source

Species:
other: three-dimensional human cornea model tissue
Strain:
other: not applicable (this is an invitro test)

Test system

Vehicle:
other: not applicable (this is an invitro test)
Controls:
other: not applicable
Duration of treatment / exposure:
3 min., 30 min. and 60 min.
Number of animals or in vitro replicates:
not applicable

Results and discussion

In vitro

Results
Irritation parameter:
other: ET50
Run / experiment:
EpiOcular Test
Value:
32.63
Negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

The test item showed the ability of direct Formazan reduction. Therefore, a functional test with freeze-killed tissues was performed and all measured values were corrected by the net formazan reduction of the freeze-killed tissues.

After 3 minutes treatment with the test item, the corrected relative absorbance values were reduced to 63.2 %. After 30 minutes treatment, corrected relative absorbance values were reduced to 57.5%. And after 60 minutes treatment, the corrected relative absorbance values were reduced to 31.9%.

From these values, the ET50was calculated mathematically. The calculated ET50of the test item Tar wood was 38.82 min.

The test item is considered as mild eye irritant.

The criterion for optical density of the negative control was fulfilled: The optical density of 1.746 is in the range of the historical data.

The positive control induced a decrease in the relative absorbance as compared to the negative control to 75.3 % for the 45 minutes treatment interval and 32.2 % for the 15 minutes treatment interval. The calculated ET50was 32.63 min., thus ensuring the validity of the test system.

Applicant's summary and conclusion

Interpretation of results:
other: mild irritant
Remarks:
Criteria used for interpretation of results: other: see table 7.5-a in the final report
Conclusions:
Tar wood is classified as mild eye irritant in the Human Cornea Model test.
Executive summary:

In the pre-test, the test item Tar wood showed the ability of direct formazan reduction. Therefore, a functional check with freeze killed tissues was performed as well.

In the main study, the test item Tar wood was applied to a three-dimensional human cornea model tissue for three different exposure times in duplicate (3 min., 30 min. and 60 min.).

100 µL of the liquid test item were applied to each tissue.

Demin. water was used as negative control, 0.3% Triton X 100 solution was used as positive control.

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 a blue formazan. Formazan production was measured by measuring the optical density (OD) of the resulting solution.

After treatment with the negative control, the absorbance values were well within the historical range and showed no cytotoxic effects. The positive control showed clear irritating effects for both treatment intervals.

After 3 minutes treatment with the test item, the relative absorbance values were reduced to 63.2 %. After 30 minutes treatment, relative absorbance values were reduced to 57.7 %. And after 60 minutes treatment, the relative absorbance values were reduced to 31.9 %. These values were calculated after subtraction of direct formazan reduction.

From these values, the ET50was calculated mathematically. The calculated ET50of the test item Tar wood was 38.82 min.

Therefore, the test item Tar wood is classified as mild eye irritant in the Human Cornea Model test.