1-Tetradecanaminium, N,N,N-trimethyl-, ethanedioate (2:1)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

An Ames test according to OECD 471 was performed with tester strains S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 with and without metabolizing system.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004-02-18 to 2004-03-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented guideline conform, scientific GLP report

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Supplemented liver extract (S9-mix) was obtained from rats which were pretreated with phenobarbital/ß-naphthoflavone, inducers of several drug metabolizing enzymes.

Test concentrations with justification for top dose:

first plate incorporation test:
a: with metabolic activation: 50, 160,500, 1600 and 5000 µg/plate
b: without metabolic activation: 50, 160, 500, 1600 and 5000 µg/plate

second plate incorporation test:
a: with metabolic activation: 0.5, 1.6,5,16,50 and 160 µg/plate
b: without metabolic activation: 0.16,0.5,1.6,5,16 and 50 µg/plate (TAI00)
0.5, 1.6,5, 16,50 and 160 µg/plate (TAI535, TA1537, TA98, WP2uvrA)

preincubation test:
a: with metabolic activation:
0.16,0.5,1.6,5,16 and 50 µg/plate (TA100, TA1537)
0.5,1.6,5,16,50 and 160 µg/plate (TA1535, TA98, WP2uvrA)
b: without metabolic activation:
0.16,0.5,1.6,5,16 and 50 µg/plate (TA100, TA1535, TA1537)
0.5, 1.6,5, 16,50 and 160 µg/plate (TA98, WP2uvrA)

Vehicle / solvent:

water

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Remarks:

Positive controls with metabolic activation for all strains: 2-aminoanthracene

Evaluation criteria:

Criteria for a valid assay:
The assay is considered valid if the following criteria are met:
- the solvent control data are within the laboratory's normal control range for the spontaneous mutant frequency
- the positive controls induce increases in the mutation frequency which are significant and within the laboratory's normal range

Criteria for a positive response:
A test compound is classified as mutagenic if it has either of the following effects:
a) it produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial background lawn
b) it induces a dose-related increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.
If the test substance does not achieve either of the above criteria, it is considered to show no evidence of mutagenic activity in this system.

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

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

The results lead to the conclusion that the test substance is not mutagenic in these bacterial test systems either in the absence or in the presence of an exogenous metabolizing system.

Executive summary:

The test substance was tested for mutagenicity according to OECD guideline 471with the strains TA 100, TA 1535, TA 1537 and TA 98 of Salmonella typhimurium and with Escherichia coli WP2uvrA.

Three independent mutagenicity studies were conducted (two plate incorporation tests, due to high toxicity and one preincubation test), each in the absence and in the presence of a metabolizing system derived from a rat liver homogenate.

For all studies, the test substance was dissolved in deionized water and each bacterial strain was exposed to 5 dose levels in the first plate incorporation test and to 6 dose levels in subsequent tests.

The concentrations used for the first plate incorporation test were 50, 160, 500, 1600 and 5000 µg/plate. This assay had to be repeated with lower concentrations because of toxic effects of the test compound resulting in less than 5 evaluable doses. Dose ranges for the second plate incorporation test and in the preincubation test were modified in individual bacterial strains to account for varying susceptibilities to cytotoxic effects. Low dose levels ranged from 0.16 to 50 µg/plate, and high dose levels ranged from 0.5 to 160 µg/plate. The test substance did not precipitate on the plates up to the highest investigated dose of 5000 µg/plate.

Mutagenicity:

In the presence and in the absence of the metabolic activation system the test substance did not result in relevant or dose-dependent increases in the number of revertants in any of the bacterial strains.

In conclusion, the test substance is not mutagenic in this bacterial mutation test at any dose level either in the absence or presence of an exogenous metabolic activation system.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

The test substance was tested for mutagenicity according to OECD guideline 471with the strains TA 100, TA 1535, TA 1537 and TA 98 of Salmonella typhimurium and with Escherichia coli WP2uvrA.

Three independent mutagenicity studies were conducted (two plate incorporation tests, due to high toxicity and one preincubation test), each in the absence and in the presence of a metabolizing system derived from a rat liver homogenate.

For all studies, the test substance was dissolved in deionized water and each bacterial strain was exposed to 5 dose levels in the first plate incorporation test and to 6 dose levels in subsequent tests. Low dose levels ranged from 0.16 to 50 µg/plate, and high dose levels ranged from 0.5 to 160 µg/plate. The test substance did not precipitate on the plates up to the highest investigated dose of 5000 µg/plate.

In the presence and in the absence of the metabolic activation system the test substance did not result in relevant or dose-dependent increases in the number of revertants in any of the bacterial strains.

Justification for selection of genetic toxicity endpoint
Well documented guideline conform, scientific GLP report

Justification for classification or non-classification

In conclusion, the test substance is not mutagenic in this bacterial mutation test at any dose level either in the absence or presence of an exogenous metabolic activation system.