Benzyldimethyl(octadecyl)ammonium chloride

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 November 08, 2017 to November 10, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Benzyldimethyl(octadecyl)ammonium chloride

EC Number:

204-527-9

EC Name:

Benzyldimethyl(octadecyl)ammonium chloride

Cas Number:

122-19-0

Molecular formula:

C27H50N.Cl

IUPAC Name:

.

Test material form:

solid

Specific details on test material used for the study:

Identity: Benzyldimethyl(octadecyl)ammonium chloride
Batch no: 15817
Composition: 100%
Cationic activity: 96.2%
Appearance: White solid
Dilution: Not diluted
Expiry date: 09/2019
Storage conditions: Room temperature, protect from humidity and water

In vitro test system

Test system:

other: SkinEthic Reconstructed Human Epidermis

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Not specified

Justification for test system used:

The SkinEthicTM skin irritation test method is a validated alternative test procedure to the Draize Rabbit Skin irritation Test and endorsed by OECD Guideline 439.

Vehicle:

unchanged (no vehicle)

Details on test system:

SkinEthicTM Reconstructed Human Epidermis:

SkinEthicTM Reconstructed Human Epidermis is a three dimensional reconstructed human skin model comprising freshly isolated normal epidermal keratinocytes (NHEK: the major cellular component of skin), which are cultured at the air liquid interface on specialised culture inserts to develop a stratified differentiated epidermis characteristic of skin in vivo. The model consist of multiple layers of viable epithelial cells and a functional multi-layered stratum corneum with robust barrier function.

The SkinEthic skin irritation assay predicts and classifies the skin irritation potential of a chemical by assessment of its cytotoxic effect following its topical application directly to the skin surface and its subsequent penetration through the stratum corneum

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

16±2 mg

Duration of treatment / exposure:

42 ± 1 minutes

Duration of post-treatment incubation (if applicable):

42 ± 1 h

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

42 minutes exposure

Value:

96.1

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

other: CLP criteria not met

Remarks:

(not irritating)

Conclusions:

Under study conditions, the test substance was determined to be non-irritant to skin

Executive summary:

An in vitro study was conducted to determine the skin irritation potential of the test substance, C18 ADBAC (active: 96.2%), using Reconstructed Human Epidermis (RHE) Test Method, according to OECD Test Guideline 439, in compliance with GLP. On Day 18, SkinEhtic^TM tissues were pre-incubated overnight in the growth culture medium. The tissues were then placed in maintenance culture medium prior to application of the test substance. 10 µL distilled water followed by 16±2 mg of powder was applied to each of the 3 RHE tissue for 42±1 minutes, then rinsed and further incubated, post exposure, for 42±1 h at 37˚C, 5% CO2±1% and 95% humidified atmosphere. Cell viability was quantitatively measured (based on dehydrogenase conversion of the MTT dye (Methylthiazolyldiphenyl-tetrazolium bromide into a blue formazan salt) by determining the absorbance at 570 nm. Sterile Dulbecco’s Phosphate Buffered Saline (DPBS) and sodium dodecyl sulphate (5% in water) were used as negative and positive controls respectively. A test substance is considered to be an irritant to skin if the skin model viability after exposure and post-treatment incubation is ≤50%. The final percentage of viability obtained with the test substance was 96.1%, which was well above the 50% irritant threshold. Optical Density (OD) results obtained with negative and positive controls were within the acceptance limits; hence the study met all the acceptance/validity criteria. Under study conditions, the test substance was determined to be non-irritant to skin (Catoire, 2017).