Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
weight of evidence
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
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
GLP compliance:
yes (incl. QA statement)
Type of study:
activation of keratinocytes

Test material

Constituent 1
Chemical structure
Reference substance name:
Bis(hydroxyethyl)methyloleylammonium chloride
EC Number:
242-332-0
EC Name:
Bis(hydroxyethyl)methyloleylammonium chloride
Cas Number:
18448-65-2
Molecular formula:
C23H48NO2.Cl
IUPAC Name:
bis(2-hydroxyethyl)(methyl)[(9Z)-octadec-9-en-1-yl]azanium chloride

Results and discussion

Positive control results:
Positive Control (Cinnamic aldehyde) induction >1.5-fold at concentration 16-128µM and EC1.5 was calculated as 8.113 µM.

In vitro / in chemico

Results
Key result
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

In this study, Bis(hydroxyethyl)methyloleylammonium chloride was classified as Negative using the KeratinoSens prediction model.

The test item did not induce statistically significant luciferase induction >1.5 in repetitions 2 and 3 but did induce significant luciferase induction in repetition 1. The respective EC1.5 value in rep 1 was calculated as 0.869 µM, however the viability was less than 70% at the inducing concentration and therefore rep 1 was also considered negative.

Sensitisation Potential of the Test Item: Repetition 1

Rep 1

Test item concentration (µM)

 

0.977

1.953

3.906

7.813

15.625

31.250

62.500

125.000

250.000

500.000

1000.000

2000.000

Mean fold induction

1.687

1.795

0.694

0.025

0.001

0.002

0.004

0.006

0.012

0.035

0.073

0.006

Viability %

56.329

19.937

-1.582

-6.962

-7.278

-5.063

0.000

-2.848

-3.797

-0.949

2.215

6.329

T-test

4.61E-06

4.08E-07

3.42E-02

2.44E-09

1.27E-09

1.28E-09

1.37E-09

1.43E-09

1.69E-09

3.22E-09

9.37E-09

1.46E-09

SD

0.078

0.220

0.156

0.019

0.000

0.000

0.001

0.001

0.003

0.008

0.021

0.002

IMAX

1.795 at 1.953 µM

EC1.5

0.869 µM

IC30

0.610 µM

IC50

1.147 µM

 

Sensitisation Potential of the Test Item: Repetition 2

Rep 2

Test item concentration (µM)

 

0.977

1.953

3.906

7.813

15.625

31.250

62.500

125.000

250.000

500.000

1000.000

2000.000

Mean fold induction

1.020

1.374

1.409

0.409

0.004

0.004

0.006

0.006

0.011

0.043

0.122

0.012

Viability %

83.740

56.325

22.708

8.848

-6.163

-5.071

-0.527

-1.368

-0.176

1.443

3.665

4.268

T-test

8.47E-01

1.17E-02

4.28E-03

8.39E-05

1.37E-09

1.37E-09

1.43E-09

1.44E-09

1.65E-09

4.02E-09

3.75E-08

1.73E-09

SD

0.275

0.380

0.229

0.143

0.001

0.000

0.001

0.001

0.002

0.014

0.051

0.005

IMAX

1.409 at 3.906 µM

EC1.5

N/A - threshold of induction not crossed at any test item concentration

IC30

1.466 µM

IC50

2.320 µM

 

Sensitisation Potential of the Test Item: Repetition 3

Rep 3

Test item concentration (µM)

 

0.977

1.953

3.906

7.813

15.625

31.250

62.500

125.000

250.000

500.000

1000.000

2000.000

Mean fold induction

0.644

0.698

0.483

0.040

0.002

0.002

0.003

0.005

0.010

0.033

0.057

0.005

Viability %

75.116

37.728

8.435

-6.187

-5.544

-4.961

1.779

-0.089

0.030

1.398

4.843

9.132

T-test

1.52E-02

4.03E-02

4.82E-04

3.73E-09

1.29E-09

1.30E-09

1.33E-09

1.40E-09

1.60E-09

3.02E-09

5.88E-09

1.40E-09

SD

0.218

0.256

0.140

0.020

0.000

0.001

0.001

0.002

0.003

0.011

0.011

0.002

IMAX

0.698 at 1.953 µM

EC1.5

N/A - threshold of induction not crossed at any test item concentration

IC30

1.111 µM

IC50

1.633 µM

Determination criteria for the skin sensitisation potential ofthe test item

 

REP1

REP2

REP3

Does at least one concentration of Test Item induce luciferase activity >1.5-fold:

Yes

No

No

Does the first concentration inducing luciferase activity above 1.5, have a viability above 70%:

No

N/A

N/A

Is p value < 0.05 for at least one concentration that yielded >1.5-fold induction with viability above 70%

Yes

N/A

N/A

Does EC1.5value occur at a concentration <1000µM (or <200µg/ml)

Yes

N/A

N/A

Does the test item induce the luciferase in a dose-dependent manner

No

N/A

N/A

Classification

Negative

Negative

Negative

All of the formal acceptance criteria of the tests were met,except for the average induction of the positive control (cinnamic aldehyde) at 32 µM, which was not within the historical range (1.6 and 3).However, as all other acceptance criteria were met, and there was a dose-dependent increase of induction with the positive control, results are considered to be valid.

Assay Acceptance Criteria (Mean of 3 repetitions)

Criteria

Result

PASS or FAIL

1

Positive Control (PC) (Cinnamic aldehyde) induction>1.5-fold in at least one concentration

16-128µM

PASS

2

Average induction of PC at 32µM is [1.6-3.0]

4.773

FAIL

3

EC1.5value is [6-39µM]

8.113

PASS

4

CV% of blank values < 20%

17.0218

PASS

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In a study conducted according to OECD Test Guideline 442D, Bis(hydroxyethyl)methyloleylammonium chloride was found to be not sensitising to the skin.
Executive summary:

The human skin sensitisation potential of Bis(hydroxyethyl)methyloleylammonium chloride was assessed using the validated in vitro method, the KeratinoSens assay, in accordance with the current OECD Test Guideline 442D. This study was assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al. (1997).

 

All of the formal acceptance criteria of the tests were met, with the exception of the average induction of the positive control (cinnamic aldehyde) at 32 µM, which was not within the historical range (1.6 and 3). However, as all other acceptance criteria were met, and there was a dose-dependent increase of induction with the positive control, the results of this study are considered to be valid.

 

The results from this study found that Bis(hydroxyethyl)methyloleylammonium chloride did not induce statistically significant luciferase induction (>1.5) in repetitions 2 or 3. Significant luciferase induction was induced in repetition 1 and the respective EC1.5 value was calculated as 0.869 µM, however, the cellular viability was less than 70% at the inducing concentration and therefore rep 1 was also considered negative. Bis(hydroxyethyl)methyloleylammonium chloride is therefore classified as Negative according to the KeratinoSens prediction model.