2-(2-methoxyethoxy)ethyl methacrylate

Endpoint summary

Description of key information

The skin corrosion potential of 2-(2-methoxyethoxy)ethyl methacrylate was assessed using a three dimensional human epidermis model in accordance with OECD TG 431 under GLP conditions. Under the conditions of the study it was concluded that 2-(2-methoxyethoxy)ethyl methacrylate is not corrosive to skin.

The skin irritation potential of 2-(2-methoxyethoxy)ethyl methacrylate was assessed using a three dimensional human epidermis model in accordance with OECD TG 439 under GLP conditions. Under the conditions of the study it was concluded that 2-(2-methoxyethoxy)ethyl methacrylate is not irritating to skin.

The eye irritation potential of 2-(2-methoxyethoxy)ethyl methacrylate was assessed in a combined a OECD EpiOcular Eye Irritation Test (EIT) and Bovine Corneal

Opacity and Permeability Test (BCOP) study under GLP. By experimental exclusion, the aggregate results of this study indicate that the test article is irritating to the eyes, but does not cause serious eye damage; therefore, 2-(2-methoxyethoxy)ethyl methacrylate is classified as GHS Category 2.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

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

Specific details on test material used for the study:

2-(2-methoxyethoxy)ethyl methacrylate

Test system:

human skin model

Source species:

human

Cell type:

other: Cultured human skin cells

Cell source:

other: Cultured human skin cells

Justification for test system used:

The EpiDerm™ Skin Model closely parallels human skin, and is recommended by test guidelines. Validation studies have reported that tests employing human skin models are able to reliably discriminate between known skin corrosives and non-corrosives.

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm™ tissues, Lot No. 27973 Kit D, were received from MatTek Corporation (Ashland, MA) on 06 Mar 2018 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.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 µl of the test article were applied to the top of each EpiDerm™ tissue. A negative control (50 µl of TCH2O) and a positive control (50 µl of KOH) were tested in the same manner. Each treatment with test article or control was conducted in duplicate.

Duration of treatment / exposure:

MatTek EpiDerm™ tissues were treated in duplicate with the test article, negative control and positive control for 3 minutes and 60 minutes

Number of replicates:

Two

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2-(2-methoxyethoxy)ethyl methacrylate (mean viability, 3 mins)

Value:

84.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2-(2-methoxyethoxy)ethyl methacrylate (mean viability, 60 mins)

Value:

68.4

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:

Skin Corrosion is defined as the production of irreversible tissue damage in skin following the application of test material. The percent viability calculated was used to determine corrosivity potential in the following manner:
Mean Viability (3 min.) < 50% = corrosive, Mean Viability (60 mins): Not applicable. From this predicted corrosivity = corrosive
Mean viability (3 min) ≥ 50 % = corrosive, Mean Viability (60 mins): < 15 % . From this predicted corrosivity = corrosive
Mean viability ( 3 min) ≥ 50 % = non-corrosive, Mean Viability (60 mins) :≥ 15 % . From this predicted corrosivity = non-corrosive

Predicted Corrosivity (60 min.) <50% Not Applicable Corrosive ≥50% <15% Corrosive ≥50% ≥15% Non-Corrosive

Interpretation of results:

GHS criteria not met

Conclusions:

2-(2-methoxyethoxy)ethyl methacrylate was not corrosive under the conditions of the OECD TG 431 in vitro study.

Executive summary:

In accordance with  OECD TG 431, using a three dimensional human epidermis model, following GLP conditions, the skin corrosion potential of 2-(2-methoxyethoxy)ethyl methacrylate is not corrosive.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

2-(2-methoxyethoxy)ethyl methacrylate

Test system:

human skin model

Source species:

human

Cell type:

other: Cultured human skin cells

Cell source:

other: Cultured human skin cells

Justification for test system used:

The EpiDerm™ Skin Irritation Model closely parallels human skin, and is recommended by test guidelines. Validation studies have reported that tests employing human skin models are able to reliably discriminate between known skin irritants and non-irritants.

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm™ tissues, Lot No. 27973 Kit D, were received from MatTek Corporation (Ashland, MA) on 06 Mar 2018 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.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

30 μl of the test article were applied to the EpiDerm™ tissue. A nylon mesh was then placed on top to facilitate even distribution of the test article.

A negative control (30 μl of PBS) and a positive control (30 μl of 5% SDS solution) were each tested concurrently, with a nylon mesh placed on top to facilitate even distribution of the material. Each treatment with test article or control was conducted in triplicate.

The exposure period for the test article and controls was 60 minutes. The dosed tissues were placed in an incubator at 37±1°C, 5±1% CO2 for 35±1 minute, and then returned to the sterile hood for the remainder of the 60-minute exposure period.

After dosing and incubation, the tissues were rinsed with PBS, blotted to remove the test substance and dry the tissue, and transferred to fresh medium. The rinsed EpiDerm™ tissues were returned to the incubator for 24±2 hours. Medium was changed at 24±2 hours. Tissues were returned to the incubator for an additional 18±2 hours.

Duration of treatment / exposure:

The exposure period for the test article and controls was 60 minutes.

Duration of post-treatment incubation (if applicable):

After dosing and incubation, the tissues were rinsed with PBS, blotted to remove the test substance and dry the tissue, and transferred to fresh medium. The rinsed EpiDerm™ tissues were returned to the incubator for 24±2 hours. Medium was changed at 24±2 hours. Tissues were returned to the incubator for an additional 18±2 hours.

At the end of the incubation period, each EpiDerm™ tissue was rinsed with PBS and transferred to a 24-well plate containing 300 μl of MTT solution (1 mg/ml MTT in DMEM). The tissues were then returned to the incubator for a three-hour MTT incubation period. Following the MTT incubation period, each EpiDerm™ tissue was rinsed with PBS and then treated with 2.0 ml of extractant solution (isopropanol) per well for at least two hours, with shaking, at room temperature. Two, 200 µl aliquots of the extracted MTT formazan were measured at 540 nm using a plate reader (μQuant Plate Reader, Bio-Tek Instruments, Winooski, VT).

Number of replicates:

Three

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2-(2-methoxyethoxy)ethyl methacrylate

Value:

61.1

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:

MatTek EpiDerm™ tissue samples were treated in triplicate with the test article, negative control, and positive control for 60 minutes. Following treatment and subsequent incubation time, the viability of the tissues was determined using thiazolyl blue tetrazolium bromide (MTT) uptake and reduction. The absorbance of each sample was measured at 540 nm. The viability was then expressed as a percent of control values. If the mean tissue viability was 50% or less, the test material was classified as an irritant; if the mean tissue viability was more than 50%, the test material was classified as a non-irritant.

Interpretation of results:

GHS criteria not met

Conclusions:

2-(2-methoxyethoxy)ethyl methacrylate was not irritating to skin under the conditions of the OECD TG 439 in vitro study.

Executive summary:

In accordance with  OECD TG 439, using a three dimensional human epidermis model, following GLP conditions, the skin irritation potential of 2-(2-methoxyethoxy)ethyl methacrylate is not irritating to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22/03/2018-21/06/2018

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 i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

0.75 ml

Duration of treatment / exposure:

10 minutes exposure

Duration of post- treatment incubation (in vitro):

- two hours after anterior and posterior chambers of the holders were then refilled with fresh MEM solution
- 90±5 minutes with sodium fluorescein solution
- Each holder was then returned to the 32±1°C incubator in a horizontal position (anterior side up) ensuring
contact of the fluorescein with the cornea for 90 minutes

Number of animals or in vitro replicates:

3

Irritation parameter:

in vitro irritation score

Value:

11.33

Negative controls validity:

valid

Remarks:

IVIS=0.17

Positive controls validity:

valid

Remarks:

IVIS=30.51

Interpretation of results:

GHS criteria not met

Conclusions:

The BCOP IVIS was 11.33, the corrected mean opacity score was 10.67, and the corrected mean optical density (permeability) score was 0.044. Because the IVIS was less than 55, this result indicated that the test article did not cause serious eye damage, thereby ruling out GHS Category 1.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22/03/2018-21/06/2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Deviations:

no

GLP compliance:

yes

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50 ul

Duration of post- treatment incubation (in vitro):

Each treatment with test article or control was conducted in duplicate. The tissues were incubated at 37±1°C, 5±1% CO2 for 30±2 minutes. After dosing and incubation, the tissues were rinsed with phosphatebuffered saline (PBS) and soaked in 5 ml of room-temperature assay medium in a 12-well plate for 12 minutes. The soaked tissues were then incubated in fresh assay medium at 37±1°C, 5±1% CO2 for 120 minutes.

Tissue Viability (MTT Reduction)
At the end of the incubation period, each EpiOcular™ tissue was transferred to a 24-well plate containing 300 μl of MTT solution (1 mg/ml MTT in Dulbecco's Modified Eagle's Medium). The tissues were then returned to the incubator for a three-hour MTT incubation period. Following the MTT incubation period, each EpiOcular™ tissue was rinsed with PBS and then treated with 2.0 ml of extractant solution (isopropanol) per well overnight at room temperature in the dark.
Two aliquots of the extracted MTT formazan from each well were transferred to a 96-well plate and optical density was measured at 570 nm using a plate reader (μQuant Plate Reader, Bio-Tek Instruments, Winooski, VT).

Number of animals or in vitro replicates:

2

Details on study design:

Test System
EpiOcular™ tissues were received and refrigerated at 2-8°C. Before use, the 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 for an additional overnight equilibration of 16 to 24 hours. After the overnight incubation, the tissues were moistened with 20 μl of phosphate-buffered saline (PBS) and incubated at 37±1°C, 5±1% CO2 for 30±2 minutes.

Irritation parameter:

other: optical density

Run / experiment:

mean

Value:

0.763

Negative controls validity:

valid

Remarks:

1.986

Positive controls validity:

valid

Remarks:

0.621

Irritation parameter:

other: tissue viability

Run / experiment:

mean [%]

Value:

38.4

Negative controls validity:

valid

Remarks:

100%

Positive controls validity:

valid

Remarks:

31.2%

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

The mean OD570 of the negative control tissues was 0.168, which met the acceptance criterion of greater than 0.8 and less than 2.5. The mean relative viability of positive control tissues was 31.2 %, which met the acceptance criterion of less than 50%. The differences in viability between identically treated tissues were 0.17% to 6.90%, which met the acceptance criterion of less than 20%. All controls passed the acceptance criteria.

In accordance with the UN GHS and OECD recommendations to perform a weight of evidence analysis of existing ex vivo and in vitro data to allow for classification of a material as an eye irritant, the combination BCOP test and EIT results were evaluated to assign the test article to a GHS acute eye hazard category.  Although neither the BCOP test, nor EIT alone can conclude GHS Category 2 classification, by taking both test together it is possible to reach a conclusion with regards to eye effects of the substance. The BCOP test has ruled out GHS Category 1 and the EIT has ruled out a non-irritant, GHS No Category, assignment. By experimental exclusion, the aggregate results of this study indicate that the test article is irritating to the eyes, but does not cause serious eye damage; therefore, the test article is classified as GHS Category 2.

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

In the EIT, the mean viability of the tissues treated with the test article was 38.4. Because tissue viability was less than 60%, the test article is irritating to the eye (GHS Category 2).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

In accordance with OECD TG 439 and OECD TG 431 conducted under GLP conditions, the respective studies concluded that 2-(2-methoxyethoxy)ethyl methacrylate is not corrosive or irritating to skin.