Reaction mass of (((1,3-phenylenebis(oxy))bis(ethane-2,1-diyl))bis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate) and (1,3-phenylenebis(oxy))bis(propane-3,1-diyl) bis(2-methylacrylate) and 3-(3-(2-(2-(methacryloyloxy)ethoxy)ethoxy)phenoxy)propyl methacrylaterate

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:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of (((1,3-phenylenebis(oxy))bis(ethane-2,1-diyl))bis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate) and (1,3-phenylenebis(oxy))bis(propane-3,1-diyl) bis(2-methylacrylate) and 3-(3-(2-(2-(methacryloyloxy)ethoxy)ethoxy)phenoxy)propyl methacrylaterate

EC Number:

944-271-1

Cas Number:

2305048-54-6

Molecular formula:

not applicable for multi-constituent.

IUPAC Name:

Reaction mass of (((1,3-phenylenebis(oxy))bis(ethane-2,1-diyl))bis(oxy))bis(ethane-2,1-diyl) bis(2-methylacrylate) and (1,3-phenylenebis(oxy))bis(propane-3,1-diyl) bis(2-methylacrylate) and 3-(3-(2-(2-(methacryloyloxy)ethoxy)ethoxy)phenoxy)propyl methacrylaterate

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: SHBH4983V
- Purity, including information on contaminants, isomers, etc.: No data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature.
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Stable for the duration of the study.
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: No data.
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: No data.
- Reactivity of the test material with the incubation material used (e.g. plastic ware): No data.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): 100% ethanol for liquid test articles, and imidazole (20%
solution in 0.9% saline) for powder or solid test articles.
- Preliminary purification step (if any): None.

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Cattle
- Number of animals: 12 corneas.
- Characteristics of donor animals (e.g. age, sex, weight): at least six months old.
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Bovine eyes (at least six months old) will be obtained from an abattoir and will be
transported to the laboratory containing Hanks’ Balanced Salt solution Hanks’
Balanced Salt Solution (HBSS) and penicillin-streptomycin in a refrigerated container.
- Time interval prior to initiating testing: No data.
- Indication of any existing defects or lesions in ocular tissue samples: The eyes will be examined after receipt from the abattoir. Any eye with a cornea
exhibiting evidence of vascularization, pigmentation, opacity or scratches will be
discarded.

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) :0.75 ml
- Concentration (if solution): 10% w/v or v/v in 0.9%

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

All corneas will be incubated at 32 (±1)°C until immediately prior to the two hour scores, at which time the MEM solution in the anterior and posterior
chambers will be removed and the holders refilled with fresh MEM solution.

Number of animals or in vitro replicates:

12

Details on study design:

NUMBER OF REPLICATES : 3 corneas per group

NEGATIVE CONTROL USED : Minimal Essential Media (MEM)

SOLVENT CONTROL USED (if applicable) : MEM solution with phenol red

POSITIVE CONTROL USED : Ethanol

APPLICATION DOSE AND EXPOSURE TIME : Surfactants are tested at a concentration of 10% w/v or v/v in 0.9% saline. A 20% (200 mg/ml) solution or suspension of the solid test article in 0.9% saline (or other vehicle as specified by the Sponsor) will be prepared using a mortar and pestle if necessary. 10 minute exposure.

TREATMENT METHOD: open chamber

POST-INCUBATION PERIOD: yes/no. If YES please specify duration

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: After 10 (±1) minutes), the test article, ethanol, or MEM was removed from the epithelium of the cornea and anterior chamber of the holder by washing with MEM solution containing phenol red. A final rinse was made with MEM without phenol red. The anterior and posterior chambers of the holders were then refilled with fresh MEM solution. Opacity measurements were made following the 10-minute exposure and MEM solution refill.
- POST-EXPOSURE INCUBATION:

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity:
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry / microtiter plate reader] (OD490)
- Others (e.g, pertinent visual observations, histopathology): (please specify)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

-0.08

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

other: See Remarks

Remarks:

Corrected Mean Optical Density

Run / experiment:

Mean

Value:

-0.005

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Run / experiment:

Mean

Value:

0

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

fluorescein leakage

Remarks:

Corneal permeability

Run / experiment:

Mean

Value:

0.016

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: CRO is technically proficient in running the OECD 437.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Range of historical values if different from the ones specified in the test guideline: Per OECD 437.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results of the test, DiBrORMA is not considered an eye irritant.

Executive summary:

The corneal irritation and damage potential of DiBrORMA was tested in the Bovine Corneal Opacity and Permeability test (BCOP). The study was performed in compliance with EPA GLP 40 CFR 160 and 792, FDA GLP 21 CFR 58, and OECD GLP (1997). The test method was based on OECD 437 (2009). A pretest was performed to measure pre-exposure opacity of the test corneas against the control corneas blanks. The corneas were prepared in cell culture and incubated at 32˚C for at least 1 hour prior to exposure. For the exposure, corneas (3/test article) were treated with 0.75 mL of undiluted liquid expressed from the test article and then were incubated at 32˚C for 10 minutes. At the end of the exposure, the corneas were washed and incubated in fresh medium for an additional 2 hours. Opacity was evaluated using an opacitometer after the 2 hour post-exposure incubation. Following opacity readings, the cell culture medium was replaced with Na-fluorescein medium and incubated for approximately 90 minutes. Following the 90 minute exposure, permeability was measured. The mean in vitro irritation score (IVIS) was calculated and the scores were classified according to protocol defined categories. Ocular exposure to DiBrORMA in the test resulted in an IVIS = -0.08, the mean opacity was 0.0 and the mean permeability score was 0.016. Based on the results of the test, DiBrORMA is not considered an eye irritant.