6-{4-[(1E)-3-methoxy-3-oxoprop-1-en-1-yl]phenoxy}hexyl methacrylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Sept. 2016 - Apr. 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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:

July 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

solid: crystalline

Details on test material:

- State of aggregation: Non-aggregated
- Particle size distribution: D50 = 12.2 µm
- Geometric standard deviation (GSD): 0.2
- Shape of particles: not assessed
- Surface area of particles: not determined
- Crystal structure: Crystalline, no further details known
- Coating: None
- Moisture content: 0.03%
- Residual solvent: Non detected
- Activation: Not applicable
- Stabilisation: With Phenothiazine (< 1000 ppm)
- Other:

Specific details on test material used for the study:

Designation in Test Facility: 16082904G
Date of Receipt: 29. Aug. 2016
Condition at Receipt: Room temperature, in proper conditions
Name: ROC-601
Batch no.: S23PSG0816
Appearance: White crystalline powder
Composition: See Certificate of Analysis
Purity: 99.5% by LC-UV
Homogeneity: Uniform crystalline powder
Expiry date: Jul. 2017
Storage: Room temperature (20 ±5 °C), keep away from light
CAS No.: 439661-46-8
EINECS-No.: Not yet assigned
Chemical Class: Organic substance: Para substituted cinnamic acid ester
Volatility: Unknown
pH-value: Unknown
LogP: Approx. 5.3 (KOWWIN v. 1.67 estimate)
Molecular weight: 346.42 g/mol
Stability: H2O: unknown; EtOH: unknown; acetone: unknown; CH3CN: unknown; DMSO: unknown
SMILES Code: C(=O)(C(=C)C)OCCCCCCOc1ccc(C=CC(=O)OC)cc1
Storage: The test item was stored in the test facility in a closed vessel at room temperature (20 ±5 °C).

Test animals / tissue source

Species:

human

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The EpiOcular(TM) tissue consists of normal, human-derived 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. These cells are not transformed or transfected with genes to induce an extended life span. The EpiOcular(TM) tissues are cultured in specially prepared cell culture inserts with a porous membrane through which nutrients can pass to the cells. The tissue surface is 0.6 cm². EpiOcular(TM) tissues were procured from MatTek In Vitro Life Science Laboratories, Mylnské Nivy 73, 82105 Bratislava, Slovakia (Designation of the kit: OCL-212-EIT, Day of delivery: 11. Oct. 2016, Batch no.: 23738).

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Tissue 1: 50.4 mg
Tissue 2: 50.7 mg

Duration of treatment / exposure:

6 hours at 37 ±1 °C, 5 ±1% CO2 and 80 – 100% relative humidity

Duration of post- treatment incubation (in vitro):

For post-treatment incubation, the tissues were incubated for 18 hours and 3 minutes at 37 ±1 °C, 5 ±1% CO2 and 80 – 100% relative humidity

Number of animals or in vitro replicates:

2 replicates

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

other: % viability

Run / experiment:

Tissue #1

Value:

101.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

other: % viability

Run / experiment:

Tissue #2

Value:

98.3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

other: % viability

Run / experiment:

Mean of two tissues

Value:

99.8

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Criterion (Demanded): Found
OD of negative control (≥ 0.8 and ≤ 2.5): 2.1
% Formazan production of positive control (< 50% of negative control): 25.8%
Variation within replicates (< 20%): 2.2% (negative control), 1.5% (positive control), 2.9% (test item)

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the test, ROC-601 is considered as not eye irritant in the EpiOcular(TM) Eye Irritation Test.

Executive summary:

In a study titled “Determination of Eye Irritation Potential ofROC-601using the EpiOcular^TM Human Cornea Model following OECD 492” the test substance ROC-601 was investigated for eye irritation potential.

One valid experiment was performed. The test item ROC-601 was applied to a three-dimensional human cornea tissue model in duplicate for an exposure time of 6 hours.

The solid test item was applied to each tissue.

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, methyl acetate was used as positive control.

The controls showed the following results: After treatment with the negative control, the absorbance values were within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.5, OD was 2.1. The positive control showed clear eye irritating effects, the relative absorbance value was reduced to 25.8 % (< 50%).

Variation within tissue replicates was acceptable (< 20%).

After treatment with the test item, the relative absorbance values were reduced to 99.8 %. This value is well above the threshold for eye irritation potential (≤ 60%).

Thus, under the conditions of the test, ROC-601 is considered as not eye irritant in the EpiOcular^TM Eye Irritation Test.