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Toxicological information

Skin irritation / corrosion

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
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 is50%

Category 2 or Category 1

Mean tissue viability > 50%

Non-Irritant