2-Propenoic acid, 2-methyl-, C13-15-branched and linear alkyl esters

Skin irritation / corrosion

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

26 June 2020

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-Propenoic acid, 2-methyl-, C13-15-branched and linear alkyl esters

EC Number:

292-124-9

EC Name:

2-Propenoic acid, 2-methyl-, C13-15-branched and linear alkyl esters

Cas Number:

90552-04-8

Molecular formula:

C(17-19)H(32-36)O2

IUPAC Name:

2-Propenoic acid, 2-methyl-, C13-15-branched and linear alkyl esters

Test material form:

liquid

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: 0023067499

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia containing: 24 EPI-200 tissues

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Epiderm 200 kit, EPI-200
- Tissue batch number(s): 34145

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 25 minutes at room temperature and for 35 minutes in the incubator at 37°C
- Temperature of post-treatment incubation (if applicable): 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: washing occurred after incubation and post-incubation

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 ml
- Incubation time: 3 hours
- Spectrophotometer: SunriseTM Absorbance Reader
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: three tissues are treated in parallel. The inter-tissue variability is considered to be acceptable if the SD of % viability is >= 18%.
- Negative and positive controls: OD570 of the negative control between 0.8 and 2.8 and of the positive control (5% SDS) ≤ 20%
- Barrier function: lower acceptance limit: ET50 = 4.0 hours; upper acceptance limit: ET50 = 8.7 hours; both for Triton X-100 (1%)
Further information are given below in the section "Any other information"

NUMBER OF REPLICATE TISSUES: 3

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test chemical is identified as requiring classification and labelling according to UN GHS (Category 2 or Category 1) if the mean percent tissue viability after exposure and posttreatment incubation is less than or equal (≤) to 50%.
- The test substance is considered to be non-corrosive to skin if after exposure is greater than or equal to 50%.
- In case of borderline results such as non-concordant replicate measurements and/or mean percent tissue viability equal to ± 5% of the cut-off value, a second test should be considered as well as a third one in case of discordant results between the first two tests.
- Justification for the selection of the cut-off point:
A borderline range (50 ± 5%) was statistically determined by using historic BASF data and hence considers the variance of the test method. This evaluation is confirming the borderline range provided in OECD Guideline 439.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 μL undiluted test substance were applied using a pipette

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration (if solution): 5% (w/v)

Duration of treatment / exposure:

25 minutes at room temperature and for 35 minutes in the incubator at 37°C

Duration of post-treatment incubation (if applicable):

18 ± 2 hour

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean value from three tissues out of one experiment

Value:

101.1

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction: The test substance ist not able to reduce MTT directly.

Any other information on results incl. tables

Individual and mean OD570 values, individual and mean viability values, standard deviations and coefficient of variation

Test substance identification		Tissue 1	Tissue 2	Tissue 3	Mean	SD	CV[%]
NC	mean OD570	1.879	1.950	2.030	1.953
	viability [% of NC]	96.2	99.9	104.0	100.0	3.9	3.9
Test substance	mean OD570	2.089	1.880	1.957	1.975
	viability [% of NC]	106.9	96.3	100.2	101.1	3.4	5.3
PC	mean OD570	0.061	0.052	0.052	0.055
	viability [% of NC]	3.1	2.7	2.6	2.8	0.3	9.7

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results observed and by applying the evaluation criteria, it was concluded that 2-Propenoic acid, 2-methyl-, C13-15-branched and linear alkyl esters does not show a skin irritation potential in the EpiDerm™ in vitro skin irritation test under the test conditions chosen.