Ethyl 2-methyl-1,3-dioxolane-2-acetate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 July 2013 to 15 July 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl 2-methyl-1,3-dioxolane-2-acetate

EC Number:

229-114-0

EC Name:

Ethyl 2-methyl-1,3-dioxolane-2-acetate

Cas Number:

6413-10-1

Molecular formula:

C8H14O4

IUPAC Name:

ethyl 2-(2-methyl-1,3-dioxolan-2-yl)acetate

Details on test material:

- Name of test material (as cited in study report): Methyl Dioxolan
- Appearance: Clear colourless liquid
- Storage condition of test material: Room temperature in the dark

In vitro test system

Test system:

human skin model

Remarks:

EPISKIN™

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: collagen type I matrix coated with type IV collagen

Vehicle:

unchanged (no vehicle)

Amount/concentration applied:

10 µL of test material was applied to the epidermis surface.

Duration of treatment / exposure:

15 minutes.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

95.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2

Value:

83

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3

Value:

103.6

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:

- Direct MTT Reduction
The MTT solution containing the test material did not turn blue which indicated that the test material did not directly reduce MTT.

- Test Material, Positive Control Item and Negative Control Item
The individual and mean OD562 values, standard deviations and tissue viabilities for the test material, negative control and positive control are given in Table 1. The mean viabilities and standard deviations of the test material and positive control, relative to the negative control are also given in Table 1.

The relative mean viability of the test material treated tissues was 94.0 % after a 15-minute exposure period and 42 hours post-exposure incubation period.

It was considered unnecessary to perform IL-1α analysis as the results of the MTT test were unequivocal.

Any other information on results incl. tables

Table 1: Mean OD₅₆₂ Values and Percentage Viabilities for the Negative Control Item, Positive Control Item, and Test Material

Substance	OD562 of Tissues	Mean OD562 of Triplicate Tissues	± SD of OD562	Relative Individual Tissue Viability (%)	Relative Mean Viability (%)	± SD of Mean Relative Viability (%)
Negative Control Item	1.006	0.842	0.144	119.5	100*	17.1
	0.787			93.5
	0.734			87.2
Positive Control Item	0.113	0.080	0.028	13.4	9.5	3.4
	0.067			8.0
	0.061			7.2
Test material	0.804	0.792	0.087	95.5	94.0	10.4
	0.699			83.0
	0.872			103.6

* = The mean viability of the negative control tissue is set at 100 %

SD = Standard Deviation.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the test the test material was considered to be a non-irritant.

Executive summary:

A test was carried out to evaluate the skin irritation potential of the test material in vitro, using the EPISKIN™ reconstructed human epidermis model. The study was conducted under GLP conditions and in accordance with the standardised guidelines OECD 439 and EU Method B.46.

The test consisted of a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours. The principle of the assay is based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test material by means of the colourimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt to a blue formazan salt (with the mitochondria of viable cells) in the test material treated tissues relative to the negative controls.

The relative mean viability of the test material treated tissues was 94.0 % after the 15-minute exposure period and 42 hour post-exposure incubation period.

As a result of this, the test material is not considered to be classified as a skin irritant according to EU criteria.