[2-[(2-methyl-1-oxoallyl)oxy]ethyl] hydrogen succinate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

[2-[(2-methyl-1-oxoallyl)oxy]ethyl] hydrogen succinate

EC Number:

244-096-4

EC Name:

[2-[(2-methyl-1-oxoallyl)oxy]ethyl] hydrogen succinate

Cas Number:

20882-04-6

Molecular formula:

C10H14O6

IUPAC Name:

4-{2-[(2-methylprop-2-enoyl)oxy]ethoxy}-4-oxobutanoic acid

impurity 1

Reference substance name:

2-hydroxyethyl methacrylate

EC Number:

212-782-2

EC Name:

2-hydroxyethyl methacrylate

Cas Number:

868-77-9

Molecular formula:

C6H10O3

IUPAC Name:

2-hydroxyethyl methacrylate

impurity 2

Reference substance name:

Succinic acid

EC Number:

203-740-4

EC Name:

Succinic acid

Cas Number:

110-15-6

Molecular formula:

C4H6O4

IUPAC Name:

succinic acid

Test material form:

liquid

Details on test material:

- Name as cited in the report: Methacryloyloxyethyl succinate
- EC-name: [2-[(2-methyl-1-oxoallyl)oxy]ethyl] hydrogen succinate
- CAS: 20882-04-6
- EC-No.: 244-096-4
- Batch No.: 3903/64
- Storage: 4-8°C, protected from heat and sunlight
- Description: yellowish liquid
- Density: 1.1884 g/kg
- Molecular weight: approx. 230 g/mol
- Formula: C10H14O6
- Purity: 84% (NMR)

In vitro test system

Test system:

human skin model

Remarks:

EpiSkin-SM (TM)

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: normal (non-cancerous), adult human-derived epidermal keratinocytes (NHEK)

Source strain:

not specified

Justification for test system used:

This test uses the EPISKIN-SM reconstructed human epidermis model (SkinEthic) which consists of normal human epidermal keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis.
This in vitro method allows Ihe identification of corrosive and non-corrosive subslances and mixtures in accordance with UN GHS. It further allows a partial sub-calegorisation of corrosives in category 1A and 1B/C .

Vehicle:

unchanged (no vehicle)

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

The test item was applied undiluted. 50 ± 3 µL (131.6 µL/cm2) of the test item was dispensed directly atop the EPISKIN-SM (TM) tissue using a positive displacement pipette. The test item was spread to match size of the tissue.

Duration of treatment / exposure:

3 min, 60 min, 4 h

Duration of post-treatment incubation (if applicable):

3 h MTT incubation

Number of replicates:

duplicate cultures per test

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean test item 3 min treatment

Value:

136

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0

Positive controls validity:

not applicable

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean test item 60 min treatment

Value:

78

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0

Positive controls validity:

not applicable

Results 3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean test item 4 h treatment

Value:

96

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0

Positive controls validity:

valid

Remarks:

5.0

Any other information on results incl. tables

PRE-EXPERIMENTS

The mixture of 50 µL test item per 2 mL MTT medium showed no reduction of MTT as compared to the solvent. The mixture did not turn blue/purple. Therefore, NSMTT equaled 0%.

The mixtures of 10 µL test item per 90 µL A. dest. and per 90 µL isopropanol showed no colouring as compared to the solvent. Therefore NSC equaled 0%.

TEST RESULTS

The test item showed no non-specific MTT-reducing or water-colouring potential, therefore no additional controls were necessary.

The test item showed no corrosive effects. The mean relative tissue viability (% negative control) was >= 35% (96%) after 4 h treatment.

Relative mean tissue viability was reduced to 78% after 60 min treatment and to 136% after 3 min treatment.

The controls confirmed the validity of the study. The mean OD570 of the two negative control tissues was between 0.6 and 1.5 for each exposure period. The mean relative tissue viability (% negative control) of the positive control was <= 20% (5%) after 4 h treatment. The maximum inter tissue viability difference of replicate tissues of all dose groups was <= 30% (0.2% - 10.0%).

3 MIN EXPERIMENT	Negative Control	Test Item	Positive Control
total mean OD570 (mean 2 replicates, blank corrected)	0.774	1.050	n.a.
mean rel. tissue viability [%]	100	136	n.a.
mean inter tissue viability diff. [%]	0.2	10.0	n.a.

60 MIN EXPERIMENT	Negative Control	Test Item	Positive Control
total mean OD570 (mean 2 replicates, blank corrected)	0.857	0.666	n.a.
mean rel. tissue viability [%]	100	78	n.a.
mean inter tissue viability diff. [%]	1.9	8.4	n.a.

4 h EXPERIMENT	Negative Control	Test Item	Positive Control
total mean OD570 (mean 2 replicates, blank corrected)	0.610	0.586	0.030
mean rel. tissue viability [%]	100	96	5
mean inter tissue viability diff. [%]	3.4	3.9	1.3

Applicant's summary and conclusion

Interpretation of results:

other: not corrosive

Remarks:

Criteria used for interpretation of results: other: OECD 431

Conclusions:

In this study under the given conditions the test item showed no corrosive effects. The relative mean tissue viability after 4 h treatment was not decreased to less than 35% of the corresponding negative control tissues. The test item is therefore classified as "non-corrosive".

Executive summary:

In the present study the skin corrosivity potential of Methacryloyloxyethyl succinate was analysed. Since corrosive chemicals are cytotoxic after topical short-time exposure to the EPISKIN-SM (TM), a reconstituted three-dimensional human epidermis model, the cytotoxic effects of the test item were determined. Cytotoxicity is expressed as the reduction of mitochondrial dehydragenase activity measured by formazan production from MTT after a 3 min, 60 min and 4 h exposure period and compared to those of the concurrent negative controls.

In this study under the given conditions the test item showed no corrosive effects. The relative mean tissue viability after 4 h treatment was not decreased to less than 35% of the corresponding negative control tissues. The test item is therefore classified as "non-corrosive".