2-(oxolan-2-ylmethoxy)ethanol

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 - 11 Aug 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

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2015

Deviations:

yes

Remarks:

no demonstration of the technical proficiency in the report

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2012

Deviations:

yes

Remarks:

no demonstration of the technical proficiency in the report

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayrisches Landesamt für Gesundheit und Lebensmittelsicherheit, Schwabach, Germany

Test material

Test material information

Constituent 1

Reference substance name:

2-[(oxolan-2-yl)methoxy]ethan-1-ol

EC Number:

608-659-1

Cas Number:

31692-85-0

Molecular formula:

C5H10O2[C2H4O]n, n = 0, 1, 2, 3, 4, ... (data given for max n of 4)

IUPAC Name:

2-[(oxolan-2-yl)methoxy]ethan-1-ol

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: normal human epidermal keratinocytes (NHEK)

Source strain:

other: 00267

Justification for test system used:

This test uses the EpiDerm™ reconstructed human epidermis model (MatTek) 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, i.e. the epidermis.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™-Standard Model (EPI-200™)
- Tissue batch number(s): 25835
- Date of initiation of testing: 09 Aug 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1 °C (35 ± 1 min) and room temperature (25 min)
- Temperature of post-treatment incubation: 37 ± 1 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: 19 washing steps: The tissues were washed by filling and emptying the inserts 15 times with DPBS using a constant stream in about 1.5 cm distance from the tissue surface, staggered again in e.g. one-minute intervals. Subsequently, the inserts were completely submerged three times in 150 mL DPBS and shaken to remove rests of the test item. Finally, the inserts were rinsed once from the inside and the outside with sterile DPBS.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration:1 mg/mL
- Incubation time: 3 h (± 5 min)
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.474 ± 0.093 (OD) (acceptance criteria (OD): 1.0 - 3.0)
- Barrier function: ET-50: 5.93 h (acceptance criteria: 4.77 - 8.72 h)
- Morphology: functional stratum corneum, a viable basal cell layer and intermediate spinous and granular layers
- Contamination: no
- Reproducibility: yes

NUMBER OF REPLICATE TISSUES: 3

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritant to skin in accordance with regulation EC 1272/2008, if the tissue viability after exposure and post-incubation is less or equal to 50% of the negative control.
- The test substance is considered to be non-irritant to skin if the tissue viability after exposure and post-treatment incubation is more than 50% of the negative control.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 30 µL
- Concentration: undiluted

NEGATIVE CONTROL
- Amount(s) applied: 30 μL

POSITIVE CONTROL
- Amount(s) applied: 30 μL
- Concentration: 5 %

Duration of treatment / exposure:

60 ± 1 min

Duration of post-treatment incubation (if applicable):

42 ± 2 h

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

(mean value)

Run / experiment:

test substance

Value:

93.6

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

Remarks:

(mean value)

Run / experiment:

positive control

Value:

3.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: not reported
- Direct-MTT reduction: No MTT reduction was observed.
- Colour interference with MTT: No colour interference was observed.

ACCEPTANCE OF RESULTS:
The mean OD value of the three negative control tissues should be between 0.8 and 2.8.
The acceptable mean percentage viability range for positive controls is ≤ 20%.
The standard deviation value (SD) calculated from individual % viabilities of the three identically treated replicates should be ≤ 18%.

- Acceptance criteria met for negative control: yes, as the mean OD value of the three negative control tissues was 1.5.
- Acceptance criteria met for positive control: yes, as the mean OD value obtained for the positive control was 0.059 and this result corresponds to 3.9 % viability compared to the results of the negative controls and thus relative viability was ≤ 20%.
- Acceptance criteria met for variability between replicate measurements: yes, as each calculated standard deviation value (SD) for the % viability was below 18% (0.4 - 4.8%).




:

Any other information on results incl. tables

Table 2: Results of MTT assay

	Tissue	OD550 (blank-corrected)		Mean OD550 (blank-corrected)	Total mean OD550 (blank-corrected)	SD OD550	Relative tissue viability [%]	Mean relative tissue viability [%]	SD tissue viability [%]	CV [% viabilities]
Negative control	1	1.592	1.552	1.572	1.500*	0.062	104.8	100	4.2	4.2
	2	1.466	1.468	1.467			97.8
	3	1.455	1.467	1.461			97.4
Positive control	1	0.062	0.066	0.064	0.059	0.005	4.3	3.9	0.4	8.9
	2	0.053	0.055	0.054			3.6
	3	0.059	0.059	0.059			3.9
Test substance	1	1.396	1.315	1.355	1.404	0.072	90.4	93.6	4.8	5.2
	2	1.401	1.336	1.369			91.3
	3	1.464	1.510	1.487			99.2

 

*         Blank-corrected mean OD₅₇₀nm of the negative control corresponds to 100% absolute tissue viability.

Applicant's summary and conclusion

Interpretation of results:

other: CLP/ EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008

Conclusions:

An in vitro skin irritation study according to OECD Guideline 439 was performed with the test substance, showing no irritation potential. Thus the test substance does not meet the classification criteria according to Regulation (EC) 1272/2008, and its data are therefore conclusive but not sufficient for classification.