Heavy wax residue fraction, waste plastic derived, hydrothermal conversion

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 3 Jan 2022 to 13 January 2022

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Heavy wax residue fraction, waste plastic derived, hydrothermal conversion

Molecular formula:

Not applicable - UVCB

IUPAC Name:

Heavy wax residue fraction, waste plastic derived, hydrothermal conversion

Test material form:

other: solid - lumps

Details on test material:

- Stability: stable under storage conditions
- Storage: room temperature (20 ± 5 °C)
- Appearance: large, brown, solid lumps
- Purity: not applicable, UVCB
- Homogeneity: homogeneous
- EC no.: 950-445-8

Test animals / tissue source

Details on test animals or tissues and environmental conditions:

The EpiOcularTM 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 EpiOcularTM 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.

EpiOcularTM tissues were procured from MatTek In Vitro Life Science Laboratories, Mlynské Nivy 73, 82105 Bratislava, Slovakia.
Designation of the kit: OCL-200-EACH
Day of delivery: 11. Jan. 2022
Batch no.: 34948

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

The solid, wax-like test item was grated with a square grater to achieve most possible shredding and fine test item fragments.

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Amount test item:
Tissue 1: 51.0 mg
Tissue 2: 51.3 mg

Duration of treatment / exposure:

6 hours

Duration of post- treatment incubation (in vitro):

18 hours

Number of animals or in vitro replicates:

2 replicates for test item, negative control and positive control

Details on study design:

Study was performed according to the guideline.

Doses used: 50 µl of controls, 51.0 mg and 51.3 mg of test item

Demonstration of proficiency:
The validity of the EpiOcularTM test at LAUS GmbH was demonstrated in a non-GLP profi-ciency study. For this purpose 15 proficiency chemicals (indicated by the OECD 492 guide-line) 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:

mean percent tissue viability 

Run / experiment:

Mean of tissue 1 and 2

Value:

98.7

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the test, Heavy wax residue fraction, waste plastic derived, hydrothermal conversion is considered non-eye irritant (GHS No Category) in the Epi-OcularTM Eye Irritation Test.

Executive summary:

The test item Heavy wax residue fraction, waste plastic derived, hydrothermal conversion 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 two tissue replicates.
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 form-azan production was evaluated by measuring the optical density (OD) of the resulting solu-tion.
Demineralized water was used as negative control and methyl acetate was used as positive control.
The controls showed the following results: After treatment with the negative control, the ab-sorbance values were within the required acceptability criterion of mean OD > 0.8 and < 2.8, OD was 1.786. The positive control showed clear eye irritating effects, the mean value of the relative tissue viability was 29.5% (< 50%).
The 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 98.7 %.
This value is above the threshold for eye irritation potential (≤ 60%). Test items that induce values above the threshold are considered non-eye irritant.