Quaternary ammonium compounds, C12-14-alkylethyldimethyl, Et sulfates

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From 15 October, 2001 to 05 March, 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study was conducted according to the OECD guideline 476, EU method B.17 as well as in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Commission directive 2002/32/EC and U.K Envoirmrntal Mutagen Society

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Coco alkyl trimethyl ammonium chloride

IUPAC Name:

Coco alkyl trimethyl ammonium chloride

Constituent 2

Reference substance name:

Quaternary ammonium compounds, coco alkyltrimethyl, chlorides

EC Number:

263-038-9

EC Name:

Quaternary ammonium compounds, coco alkyltrimethyl, chlorides

Cas Number:

61789-18-2

IUPAC Name:

61789-18-2

Test material form:

other: Pale straw colour liquid

Details on test material:

- Name of test material (as cited in study report): Arquad C-35 (purity 35%)
- Description: Pale straw colour liquid
- Batch number: RH020010082
- Storage conditions: Room temperature, in the dark

Method

Target gene:

Thymidine kinase gene

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

For Experiment 1: 0.625, 1.25, 2.5, 5, 10 and 20 µg/mL for both with and without S9-mix
For Experiment 2: 0.313, 0.625, 0.938, 1.25, 2.5 and 5 µg/mL (without S9 mix) and 2.5, 5, 10, 15, 20 and 30 µg/mL (with S9-mix)

Vehicle / solvent:

RPMI 1640 without serum

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

in the absence of S9-mix

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

in the presence of S9-mix

Details on test system and experimental conditions:

After a preliminary toxicity test, 0.625, 1.25, 2.5, 5, 10 and 20 µg/L was selected for the first experiment. The entire experiment was repeated to confirm the results of the first experiment. 3 h exposure were used both with and without S9 mix while for the second experiment, the exposure time was increased to 24 h. For the second experiment, the dose range was 2.5 to 30 µg/L with S9 mix and 0.313 to 5 µg/L without S9 mix.

Evaluation criteria:

For a test substance to give a significant result then two or more of the following criteria should be met:
a) A greater than three-fold increase in the mutant frequency/survivor over the negative control value.
b) A dose-related increase in the mutant frequency per survivor.
c) An increase in the absolute number of mutants.

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: Yes

Remarks on result:

other: other: L5178Y TK+/- mouse lymphoma cells

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

There was evidence of toxicity with the test substance in both the presence and absence of S9-mix as indicated by relative suspension growth (RSG), this was confirmed by the decrease in relative total growth (RTG). There was no evidence of a reduction in Day 2 viability, therefore indicating that no residual toxicity occurred, in either the presence or absence of S9-mix. Optimum level of toxicity was achieved, in both the presence and absence of S9-mix. The toxicity observed at 30 µg/mL in the presence of metabolic activation exceeded the upper acceptable limit of 90%; therefore it was excluded from the statistical analysis.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with or without metabolic activation

Under the test conditions, the test substance did not show mutagenic activity in mouse lymphoma assay.

Executive summary:

An OECD 476 study was conducted to evaluate the potential of the read-across substance C12-18 TMAC (active ingredient 35%) to induce mutations at the TK (thymidine kinase) locus in L5178Y mouse lymphoma cells. After a preliminary toxicity test, 0.625, 1.25, 2.5, 5, 10 and 20 µg/L was selected for the first experiment. The experiment was repeated to confirm the results of the first experiment. Three hour exposures were used both with and without S9-mix while for the second experiment, the exposure time was increased to 24 hours. For the second experiment, the concentration range was 2.5 to 30 µg/L with S9-mix and 0.313 to 5 µg/L without S9-mix. The test substance did not induce a statistically significant or concentration-related increase in the mutant frequency at any concentration level, either with or without metabolic activation. Adequate levels of toxicity were achieved in all exposure groups. Under the test conditions, C12-18 TMAC did not show mutagenic activity in mouse lymphoma assay.