Benzyldimethyl(octadecyl)ammonium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From January 30, 2018 to Febraury 02, 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 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

Adopted october 2017

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

Test animals / tissue source

Species:

human

Strain:

other: human-derived keratinocytes

Details on test animals or tissues and environmental conditions:

The EpiOcular tissue consists of normal, human-derived keratinocytes which have been cultured to form a stratified squamous epithelium similar to that found in the human cornea. It consists of highly organized basal cells. These cells are not transformed or transfected with genes to induce an extended life span. The EpiOcularTM tissues are cultured in specially prepared cell culture inserts with a porous membrane through which nutrients can pass to the cells. The tissue surface is 0.6 cm2.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50 mg

Duration of treatment / exposure:

6 h

Duration of post- treatment incubation (in vitro):

18 h

Number of animals or in vitro replicates:

Duplicates

Results and discussion

In vitro

Results

Irritation parameter:

other: % relative tissue viability

Run / experiment:

6 h

Value:

5.7

Vehicle controls validity:

not valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Any other information on results incl. tables

% Viability positive control and test substance:

Treatment	Acceptance criterion	OD570nm	Viability %	Difference of viability	% Mean viability
Water (NC)	0.8≤OD≤2.5	1.653	100.3	0.5	100
	Difference of viability <20%	1.644	99.7
Methyl acetate	%mean viability < 50%	0.173	10.5	13.2	17.1
	Difference of viability <20%	0.391	23.7
Test substance	Difference of viability <20%	0.136	8.2	5.2	5.7
		0.051	3.1

Applicant's summary and conclusion

Interpretation of results:

other: CLP criteria not met (Inconclusive)

Conclusions:

Under the conditions of the study, the test substance can be considered to be potentially irritating, however no prediction for classification (i.e., GHS class 1 and GHS class 2) can be made

Executive summary:

A study was conducted to determine the eye irritation potential of the test substance, C18 ADBAC (active: 96.2%), using the commercially available MatTek® Human Corneal Epithelium (EpiOcular) test method, according to OECD 492 Guideline, in compliance with GLP.EpiOcularTM tissues were placed in 1 mL of fresh maintenance medium (6-well plate). The tissues were incubated overnight at 37°C, 5% CO2, in humidified incubator and after 1 h, the medium was replaced by 1 mL of fresh medium for an overnight incubation at at 37°C, 5% CO2, in humidified incubator. The tissues in duplicate were exposed to single topical application of 50 mg neat test substance or 50 µL of reference substances (negative control: sterile water; positive control: methyl acetate) for 6 h, followed by a 25 minutes post-treatment immersion, and 18 h post-treatment incubation, prior to the MTT endpoint. After different treatment exposure period, tissues were transferred to 24 well plates containing MTT medium (1 mg/mL). After 3 h MTT incubation, the blue formazan salt formed was extracted with 2 mL of isopropanol per tissues for 2 to 3 h at room temperature or overnight at 4°C. The absorbance of the resulting coloured solution was measured at 570 nm. The viability of the tissues (which has been shown to directly correlate with the irritation potential) were assessed and compared to a negative control. The percentage viability obtained with the test substance was determined to be 5.7%, which is well below the threshold (i.e., ≤60%) indicating no prediction can be made. All the assay acceptance criteria were met and the study was considered valid. Under the conditions of the study, the test substance can be considered to be potentially irritating, however no prediction for classification (i.e., GHS class 1 and GHS class 2) can be made (Catoire, 2018).