Dimethylbis[2-[(1-oxohexadecyl)oxy]ethyl]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:

30/01/2018 until 01/02/2018

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:

2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test animals / tissue source

Species:

other: MatTek® Human Corneal Epithelium (EpiOcular)

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Protocol for solid test articles: approximately 50 mg of test article were applied topically on cultures for 6 hours, followed by a 25-minute post-treatment immersion, and 18-hour post-treatment incubation, prior to the MTT endpoint.

Duration of treatment / exposure:

6 hours

Duration of post- treatment incubation (in vitro):

25-minute post-treatment immersion, and 18-hour post-treatment incubation

Number of animals or in vitro replicates:

2

Results and discussion

In vitro

Results

Irritation parameter:

other: cell viability [%]

Remarks:

relative to negative control

Value:

> 60

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Assay Quality Controls
Assay Acceptance Criterion 1: Negative Control (NC)
- Negative Control meets the acceptance criterion.
Assay Acceptance Criterion 2: Positive Control (PC)
- Positive Control meets the acceptance criterion.
Assay acceptance criterion 3: Difference of viability
- The difference of viability between two tissues meets the acceptance criterion.
Tiered test for direct MTT reduction
- The MTT tiered test ensures that there is not direct interaction between the test samples and MTT and therefore corrects against chemicals which might otherwise be falsely predicted as non-irritant. If interaction is observed, the test substances are applied to freeze killed controlled tissue to determine the likelihood of the interaction occurring under test conditions.
- Dimethylbis[2-[(1-oxohexadecyl)oxy]ethyl]ammonium chloride was incubated with MTT for 180 minutes. There was no visual evidence of MTT reduction (violet coloration). Therefore application onto freeze killed tissue was not required.
Colored test chemical
- Colored test chemicals or test chemicals able to develop a color after contact with the tissue can generate a remaining Non-Specific Color on living tissues.
When a colored solution is observed, it is then necessary to evaluate during the future studies the part of OD due to the Non-Specific Color (i.e by using living epidermis tissues without MTT conversion test) to define the %NSCliving value. Nevertheless if the final result classifies the chemical as irritant, it is not necessary to subtract the %NSCliving value.
- Dimethylbis[2-[(1-oxohexadecyl)oxy]ethyl]ammonium chloride was incubated with water and isopropanol for 1hr. There was evidence of white turbidity. Nevertheless, only test chemicals that appear red, blue, black and green should be potentially considered as intrinsic colorants. Dimethylbis[2-[(1-oxohexadecyl)oxy]ethyl]ammonium chloride did not produce colored interference, only white turbidity.
Batch release quality control
- The tissue batch n°27021 met the acceptance criteria defined into the certificate of analysis.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Dimethylbis[2-[(1-oxohexadecyl)oxy]ethyl]ammonium chloride did not reduce cell viability to below 60% and was classified as non irritant for eyes according to the OECD 492 guideline.