1H-Imidazolium, 3-ethyl-1-methyl-, tetrafluoroborate(1-) (1:1)

Genetic toxicity: in vitro

Administrative data

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:

Experimental Starting date: 2015-04-07; experimental completion date: 2015-04-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

resp. EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

yes

Remarks:

The syntax of the test item name was changed, this can be seen as uncritical

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht; Kaiser-Freidrich-Straße 7, D-55116 Mainz

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

17PI120_1

IUPAC Name:

17PI120_1

Test material form:

other: viscous liquid

Details on test material:

Stabilized EMIM BF4:
Appearance: lightly yellow clear viscous liquid
Purity: 93.36% (EMIM+, HPLC); 86.68% (BF4-, titration); 4.89% (Stabilizer, HPLC)

Method

Species / strain

Species / strain / cell type:

other: 9005=S. typhimurium TA 1535, TA 97a, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

other: lipopolysaccharide side chain deficiency

Metabolic activation:

with and without

Test concentrations with justification for top dose:

1. Experiment:
5 concentrations of the test item ranging from 50 µg/plate to 5000 µg/plate using the plate incorporation method

2. Experiment
7 concentrations of the test item ranging from 78 µg/plate to 5000 µg/plate using the pre-incubation method

Vehicle / solvent:

dimethyl sulfoxide (DMSO) was used as vehicle

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: 2-Amino-anthracene; 4-Nitro-1,2-phenylene Diamine

Details on test system and experimental conditions:

Specification:
Species Salmonella typhimurium LT2
Strains TA97a, TA98, TA100, TA102 and TA1535

Origin and culture:
Salmonella typhimurium (all strains used) were obtained from TRINOVA BioChem (batch of the bacteria strains: TA97a: 4904D, TA98: 4903D, TA100: 4902D, TA102: 4872D, TA1535: 4908D) and were stored as lyophilisates in the fridge at 2-8 °C.
One day before the start of each experiment, one lyophilisat per strain to be used was taken from the fridge to inoculate a culture vessel containing nutrient broth. After incubation overnight at 37 ± 1 °C, the cultures were used in the experiment. During the test, the cultures were stored at room temperature as to prevent changes in the titre.

Evaluation criteria:

The colonies were counted visually, the numbers were recorded. A spreadsheet software (Microsoft Excel®) was used to calculate mean values and standard deviations of each treatment, solvent control and positive control.
The increase factor f(I) of revertant induction (mean revertants divided by mean spontaneous revertants) and the absolute number of revertants (“Rev. abs.”, mean revertants less mean spontaneous revertants) were also calculated.
A test item is considered to have mutagenic potential, if a significant, reproducible increase of revertant colonies per plate (increase factor  2) in at least one strain can be observed. A concentration-related in-crease over the range tested can also be taken as a sign of mutagenic activity.
Note: All calculations are performed with unrounded values. Therefore, re-calculation with rounded values may lead to slightly different results.

Results and discussion

Test results

Species / strain:

other: 4116=S. typhimurium TA 1535, TA 97a, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

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

The test item stabilized EMIM BF4 is considered as “not mutagenic under the conditions of the test”.

Executive summary:

Two valid experiments were performed.

 

First Experiment:

Five concentrations of the test item, dissolved in dimethyl sulfoxide (ranging from 50 µg/plate to 5000 µg/plate) were used. 5 genetically changed strains ofSalmonella typhimurium(TA97a, TA98, TA100, TA102 (genetically manipulated) and TA1535) were exposed to the test item both in the presence and in the absence of a metabolic activation system (S9-mix, rat liver S9-mix induced by Aroclor 1254) for 48 h at 37 ± 1°C, using the plate incorporation method.

None of the concentrations caused a significant increase in the number of revertant colonies in the tested strains. The test item did not show any mutagenic effects in the first experiment.

The test item showed no precipitates on the plates in all tested concentrations

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation.

Second Experiment:

To verify the results of the first experiment, a second experiment was performed, using 7 concentrations of the test item (ranging from 78 µg/plate to 5000 µg/plate) and a modification in study performance (pre-incubation method at 37 ± 1°C).

The test item did not show mutagenic effects in the second experiment, either.

The test item showed no precipitates on the plates in all tested concentrations.

No signs of toxicity towards the bacteria could be observed.

The sterility control and the determination of the titre did not show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation.

 

Under the conditions of the test, the test item did not show mutagenic effects towardsSalmonella typhimurium,strainsTA97a, TA98, TA100, TA102 and TA1535.

Therefore, no concentration-effect relationship could be determined.