4-allylveratrole

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 October 2017-23 November 2017

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-allylveratrole

EC Number:

202-223-0

EC Name:

4-allylveratrole

Cas Number:

93-15-2

Molecular formula:

C11H14O2

IUPAC Name:

1,2-dimethoxy-4-(prop-2-en-1-yl)benzene

Test material form:

liquid

In vitro test system

Test system:

human skin model

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE

EpiSkin Small Model kit provided by SkinEthic

- Model used: EpiDerm™ (EPI-200-SIT)
- Tissue batch number(s): 17-EKIN-044
- Delivery date: 31 October , 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: incubator 37±1°C
- Temperature of post-treatment incubation (if applicable): incubator 37±1°C, 5±1% CO2

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

10 ul

Duration of treatment / exposure:

15 min

Duration of post-treatment incubation (if applicable):

42 h

Number of replicates:

Three

Test system

Details on study design:

The test item was tested for possible skin irritation potentiial using RHE model, EpiSkin through topical application for 15 minutes. After 42 hour post-incubation perod, irritation potential of the test item was evaluated by assessing cytotoxic (irritancy) effect. Cytotoxicity is expressed as a reduction of mitochodrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

122.59

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1. Skin irritation potential of 4 -allylveratrole after 15 min exposure in reconstructed human epidermis (RHe)

Sample	OD Mean	SD of OD	Viability mean %	In vivo prediction
Negative control	0,646587	0,006166	100	NI
Positive control	0,03837	0,001103	5,93	I
Test item	0,79262	0,001477	122,59	NI

OD= optical density

S=standard deviation

NI=non-irritant

I=irritant

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item, Methyl Eugenol/4-allylveratrole, is non-irritant in the in vitro skin irritation test using reconstructed human epidermis.

Executive summary:

The test item 4 -allylveratrole/Methyl Eugenol was tested for its possible skin irritation potential using a three dimestional Reconstructed Human Epidermis model, EpiSkin. The magnitude of viability was quantified by using MTT test. Validity of the test method was ascertained by positive control 5% SDS. Three tissue replicates were used for each treatment (exposure time 15 minutes), including negative and positive control.  

The tissue viability met the acceptance criterion. Mean OD₅₇₀ of negative control was  0,646587. The viability of culture treated by positive control 5% SDS was 6.9%. The positive control met the acceptance criterion: mean tissue viability less than 20%. Determined viability of culture treated by 4 -allylveratrole/Methyl Eugenol (122,59%) fulfilled the criteria for non-irritancy. Therefore, the is considered to be non-irritant to the skin.

Criteria for interpretation	Classification UN GHS
Mean tissue viability is≤50%	Category 2 or Category 1
Mean tissue viability > 50%	Non-Irritant