[No public or meaningful name is available]

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 19 February to 05 March 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

June 26, 2020

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

N,N,N-trimethyl-3-{[(2E)-3-phenylprop-2-enoyl]amino}propan-1-aminium chloride

Molecular formula:

C15H23ClN2O

IUPAC Name:

N,N,N-trimethyl-3-{[(2E)-3-phenylprop-2-enoyl]amino}propan-1-aminium chloride

Test material form:

liquid: viscous

Remarks:

Deliquescent solid appearing as a viscous liquid.

Details on test material:

Storage: room temperature
Supplier: sponsor
batch/lot number: 209-653 (dried material from [Tradename] batch number 0001569403)
Expiry: December 3, 2021

In vitro test system

Test system:

human skin model

Remarks:

EpiDermTM (EPI-200-MatTek Corporation)

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

Initially, the predictive capacity of the modified EpiDerm™ Skin Irritation Test (SIT) test method using MatTek EpiDermTM tissue model EPI-200 underwent full prospective validation from 2003-2007. The test method components of this method were used to define the essential test methods components of the original and updated ECVAM Performance Standards (PS). A modification of the original EpiDerm™ SIT was validated using the original ECVAM PS in 2008. In 2008, ESAC concluded that the Modified EpiDerm™ SIT has sufficient accuracy and reliability for prediction of R38 skin irritating and no-label (non-skin irritating) test substances.

Vehicle:

unchanged (no vehicle)

Details on test system:

The reconstructed human epidermal model EpiDermTM (EPI-200-MatTek Corporation) was used.

MatTek’s EpiDermTM model has been extensively characterised for multiple parameters including morphology, tissue viability, skin barrier function and sterility (for information see www.mattek.com). QC results for the specific lot of models received (Lot# 34130) were checked in-house for compliance with MatTek acceptance ranges with the following outcome (details in Annex 1):

• Morphology - PASS
• Tissue viability - PASS
• Skin barrier function (ET50 value for 1% Triton X-100) where ET50 is the time taken for 1% Triton X-100 to reduce the viability of the skin model to 50% relative to the negative control)- PASS
• Sterility testing showed no contamination during long term antibiotic and antimycotic free culture- PASS

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

30 µl of neat test item, negative control (DPBS) or positive control (SDS 5%)

Duration of treatment / exposure:

60±1 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2, in humidified conditions)

Duration of post-treatment incubation (if applicable):

42±4 hours post-treatment incubation, prior to the MTT endpoint; three tissues per condition (n=3).

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Remarks:

relative to negative control

Run / experiment:

Run 1 (3 replicates) 60±1 minute exposure on the surface of the EpiDermTM reconstructed human epidermis, and a 42 ±4 hour post-exposure incubation time.

Value:

46.604

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

All acceptance criteria indicated in the method section were passed

Any other information on results incl. tables

Table: Mean and SD of cell viability measurements and of viability percentages after 60±1 minutes of application and 42±4 hours post-incubation

 

Name	Code	Mean of OD	SD of OD	Mean of viability (%)	SD of viability (%)	CV %	Classification
DPBS	NC	1.674	0.032	100.000	1.900	1.900	Non-Irritant
SDS 5%	PC	0.081	0.024	4.860	1.442	29.670	Irritant
N,N,N-trimethyl-3-{[(2E)-3-phenylprop-2-enoyl]amino}propan-1-aminium chloride	TA6	0.780	0.182	46.604	10.880	23.346	Irritant

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Remarks:

A test item is considered an irritant (I) to skin in accordance with UN GHS Category 2 if the skin model viability after exposure and post-treatment incubation is ≤50%.

Conclusions:

The percentage of viability obtained with the test item N,N,N-trimethyl-3-{[(2E)-3-phenylprop-2-enoyl]amino}propan-1-aminium chloride was 46.604%, therefore it was considered as Irritant to the skin.

Executive summary:

In this study, the skin irritation potential of N,N,N-trimethyl-3-{[(2E)-3-phenylprop-2-enoyl]amino}propan-1-aminium chloride was assessed in vitro according to OECD Test Guideline 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method.

After 60±1 minute exposure on the surface of the EpiDerm^TM reconstructed human epidermis, and a 42 ±4 hour post-exposure incubation time, viability of the tissues was assessed and compared to the negative control.

The percentage of viability obtained was 46.604% and therefore: N,N,N-trimethyl-3-{[(2E)-3-phenylprop-2-enoyl]amino}propan-1-aminium chloride was classified as Irritant to the skin.