[(2-chlorophenyl)methylene]malononitrile

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)

Version / remarks:

This study is carried out according the OECD Guideline 442D dated February, 04th, 2015, the ECVAM DBALM protocol 155: KeratinoSensTM, C(81)30 OECD Guidelines and the article annex II to article D-523-8 of the French Environment

Deviations:

not specified

GLP compliance:

yes

Remarks:

not mentioned on study reports but on IDEAtest group website www.groupeideatests.com

Type of study:

activation of keratinocytes

Justification for non-LLNA method:

Alternative study to in-vivo as the first intention, in accordance with REACH annex VII and the Adverse Outcome Pathway (AOP) for Skin Sensitisation Initiated by Covalent Binding to Proteins (OECD, 2014)

Test material

Test material information

Constituent 1

Reference substance name:

[(2-chlorophenyl)methylene]malononitrile

EC Number:

220-278-9

EC Name:

[(2-chlorophenyl)methylene]malononitrile

Cas Number:

2698-41-1

Molecular formula:

C10H5ClN2

IUPAC Name:

[(2-chlorophenyl)methylene]malononitrile

impurity 1

Reference substance name:

unknown impurities

Cas Number:

Not applicable

Molecular formula:

Not applicable

IUPAC Name:

unknown impurities

Test material form:

solid

In vitro test system

Details on the study design:

see OECD 442D guidelinae and enclosed study plan

Results and discussion

Positive control results:

Cinnamaldehyde EC1.5 7.92 and Imax 7.64

In vitro / in chemico

Results

Results 1

Key result

Run / experiment:

other: mean on 2 runs

Parameter:

other: IC70 is concentration in μM for which we obtained 70% cell viability

Value:

23.31

Vehicle controls validity:

not examined

Remarks:

control solvent as negative control

Negative controls validity:

valid

Remarks:

control solvent

Positive controls validity:

valid

Remarks:

Cinnamaldehyde Imax is 7.64

Remarks on result:

positive indication of skin sensitisation

Results 2

Key result

Run / experiment:

other: Linear EC1.5 μM and EC1.5 Lin/Log μM

Parameter:

other: EC1.5, value representing the concentration for which induction of luciferase activity is above 1.5 threshold was obtained

Value:

2.97

Vehicle controls validity:

not examined

Remarks:

constrol solvent as negative control

Negative controls validity:

valid

Remarks:

control solvent

Positive controls validity:

valid

Remarks:

cinnamaldehyde EC1.5 is 7.92 μM

Remarks on result:

positive indication of skin sensitisation

Other effects / acceptance of results:

Since all validity criteria are met, study is considered as valid.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

In both repetition, Imax is higher than 1.5, the EC1.5 is lower than 1000 μM and, at the EC1.5 concentration, viability is higher than 70%.

Under the retained experimental conditions, the test item may be classified as sensitizer.

Executive summary:

In vitro sensitization KeratinoSens™ assay OECD 442D is performed with Keratinocytes based on the signaling pathway Keap1-Nrf2-ARE coupled to the luciferase reporter gene. Under the retained experimental conditions, the test item may be classified as sensitizer.