Hexamethylene diisocyanate, oligomers, reaction products with 4-Hydroxybutyl acrylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-{[(6-{2,4,6-trioxo-3,5-bis[6-({[4-(prop-2-enoyloxy)butoxy]carbonyl}amino)hexyl]-1,3,5-triazinan-1-yl}hexyl)carbamoyl]oxy}butyl prop-2-enoate

EC Number:

929-915-1

IUPAC Name:

4-{[(6-{2,4,6-trioxo-3,5-bis[6-({[4-(prop-2-enoyloxy)butoxy]carbonyl}amino)hexyl]-1,3,5-triazinan-1-yl}hexyl)carbamoyl]oxy}butyl prop-2-enoate

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 16/438-1, 360-71-3
- pH 5
- The test substance was homogenous by visual inspection.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

Test animals / tissue source

Species:

human

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST SYSTEM
- Source: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia
- Tissue model: OCL-200
- Tissue Lot Number: 23760
- Incubation conditions: 37°C +/- 1°C, 5% +/- 1% CO2, 90% +/- 5% relative humidity
- Detection agent: MTT
- Ectracting agent: Isopropanol
- Wash buffer: PBS

Test system

Controls:

other: Negative control: deionized sterile water; positive control: Neat methyl acetate

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µl undiluted test substance

Duration of treatment / exposure:

30 minutes

Duration of post- treatment incubation (in vitro):

2h

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed with sterile PBS
- Time after start of exposure: 30 minutes

SCORING SYSTEM:
Mean tissue viability
(% of negative control) Prediction

< 55 Irritant
55 - 65 Borderline
> 65 Non-irritant

Results and discussion

In vitro

Results

Irritation parameter:

other: Tissue viability (% of NC)

Run / experiment:

Mean

Value:

93.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Any other information on results incl. tables

Individual and mean OD570 values, individual and mean viability values and inter-tissue variability

Test substance		Tissue 1	Tissue 2	Mean	Inter-tissue variability (%)
NC	Mean OD570	1.624	1.625	1.624
NC	Viability (% of NC)	100.0	100.0	100.0	0.1
Test item	Mean OD570	1.545	1.502	1.523
Test item	Viability (% of NC)	95.1	92.5	93.8	2.6
PC	Mean OD570	0.333	0.448	0.390
PC	Viability (% of NC)	20.5	27.6	24.0	7.1

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results observed for the EpiOcular Test alone and by applying the evaluation criteria it was concluded that the test item does not show an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen.

Executive summary:

The objective was to assess the eye irritating potential of the test substance. By using the methods currently available a single in vitro assay is not sufficient to cover the full range of eye irritating potential. Therefore, two in vitro assays were part of this in vitro eye irritation test strategy: The Bovine Corneal Opacity and Permeability Test (BCOP Test) and EpiOcular Eye Irritation Test.

However, in case of the test substance, the results derived with EpiOcular alone were sufficient for a final assessment. Therefore, further testing in BCOP was waived.

The potential of the test substance to cause ocular irritation was assessed by a single topical application of undiluted test substance to a reconstructed three-dimensional, human cornea model (EpiOcular™). Two EpiOcular™ tissues were incubated with the test substance for 30 minutes followed by a 2-hour post-incubation period. Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation by using a colorimetric test. The reduction of mitochondrial dehydrogenase activity measured by reduced formazan production after incubation with a

tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the epidermal tissues treated with the test substance is compared to that of negative control tissues. The ratio of the values indicates the relative tissue viability.

The following results were obtained in the EpiOcular™ eye irritation assay:

The test substance is not able to directly reduce MTT. The final mean viability of the tissues treated with the test substance was 93.8%.

Based on the results observed in the EpiOcular Test alone and by applying the evaluation criteria, it was concluded that the test substance does not show an eye irritation potential in the in vitro eye irritation test strategy under the test conditions chosen.