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

Endpoint summary

Description of key information

An in vitro skin corrosion study and an ex vivo eye irritation/corrosion study were conducted with DiBrORMA. The results of the studies were:

Not irritation when tested according to OECD 437.

Not corrosive when tested according to OECD 431.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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: 3M Company, Batch: 653940
- Purity, including information on contaminants, isomers, etc.: No data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 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: No data
- 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): test article solubilized in tissue culture water
- Preliminary purification step (if any): No data
- Final concentration of a dissolved solid, stock liquid or gel: No data

FORM AS APPLIED IN THE TEST: test article solubilized in tissue culture water

Test system:

human skin model

Remarks:

MatTek EpiDerm™ tissues

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: No data

Justification for test system used:

Per OECD 431.

Vehicle:

other: See 'Remarks'

Remarks:

Tissue culture water

Details on test system:

SKIN DISC PREPARATION
- Procedure used: EpiDerm™ tissues, Lot No. 27144 Kit O, were received from MatTek Corporation (Ashland, MA) on
03 Oct 2017 and refrigerated at 2-8°C. Before use, tissues were incubated (37±1°C, 5±1% CO2) with
assay medium (MatTek) for a one-hour equilibration. Equilibration medium was replaced with fresh
medium before dosing.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37C
- Temperature of post-treatment incubation (if applicable): No data

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: Following exposure to test materials, any test material remaining atop the EpiDerm™ tissues will be discarded. Each insert will be individually rinsed extensively with phosphate buffered saline (PBS) to remove residual test material. For a test article in which the tissue or assay medium undergoes a noticeable color change (due to pH change, staining of the tissue, etc.), an extra rinsing procedure will be performed before the MTT conversion. The tissues will be submerged twice in fresh PBS and then incubated in PBS at room temperature for 3 minutes.
- Observable damage in the tissue due to washing: No data
- Modifications to validated SOP: No data

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: visible range
- Wavelength: 540 to 570 nm
- Filter: No data
- Filter bandwidth: No data

NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the mean viability at 3 min is greater than 50% or if the mean viability is less than or equal to 50% at 3 min but the mean viability at 60 min is greater than 15%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 uL

Duration of treatment / exposure:

3 or 60 minutes

Duration of post-treatment incubation (if applicable):

None

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Remarks:

3 minutes

Value:

95.5

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

not examined

Remarks on result:

other:

Remarks:

Non-corrosive

Irritation / corrosion parameter:

% tissue viability

Remarks:

60 minutes

Value:

91.9

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

not examined

Remarks on result:

other:

Remarks:

Non-corrosive

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: None
- Direct-MTT reduction: None
- Colour interference with MTT: None detected

DEMONSTRATION OF TECHNICAL PROFICIENCY: The CRO running the study is techincally proficient in running OECD 439 with MatTek tissues.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes
- Range of historical values if different from the ones specified in the test guideline: Per OECD 431 and MatTek protocol.

Interpretation of results:

GHS criteria not met

Conclusions:

The percent viability was 95.5% at 3 minutes and 91.9% at 60 minutes. The test article was non-corrosive. 

Executive summary:

DiBrorma was tested in a GLP-compliant, OECD 431EpiDerm™ Skin Corrosivity Test.MatTek EpiDerm™ tissues were treated in duplicate with the test article, negative control, tissue culture water, and positive control, potassium hydroxide solution, for 3 minutes and 60 minutes. Following treatment, the viability of the tissues was determined using thiazolyl blue tetrazolium bromide (MTT) uptake and conversion, and the absorbance of each tissue was measured at 540 nm. The mean viability was then expressed as a percent of control values. The percent viability was used to determine corrosivity potential. The percent viability was 95.5% at 3 minutes and 91.9% at 60 minutes. The test article was non-corrosive. 

Eye irritation

Link to relevant study records

Reference

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

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

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.

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.

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)

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

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

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

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.

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

Based on the results of the eye irritation study, DiBrORMA does not meet the classification criteria for eye irritation.

DiBrORMA is not corrosive to skin, but data on skin irritation potential are not available.  Out of an abundance of caution, and because this chemical class can be irritating to skin, DiBrORMA is classified for skin irritation, GHS Category 2.