N,N-dimethyltetradecylamine N-oxide

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 based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

March 4 2010 - May 12-2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C12-14 (even numbered) -alkyldimethyl, N-oxides

IUPAC Name:

Amines, C12-14 (even numbered) -alkyldimethyl, N-oxides

Constituent 2

Reference substance name:

931-292-6

EC Number:

931-292-6

IUPAC Name:

931-292-6

Details on test material:

- Name of test material (as cited in study report): Amin, C10-16-alkyldimethyl, N-oxide (AO-1270)
- Physical state: liquid
- Analytical purity: 28-30 %w/w based on AO content
- Composition of test material, percentage of components: Amineoxide 28.00 – 30.00 %; Amine <0.100 %; Water 70 – 72 %; Hydrogen peroxide 0.02 %
- Lot/batch No.: SWF-2061-041
- Stability under test conditions: stable
- Storage condition of test material: At a dry, cool place

Method

Target gene:

HPRT locus

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: yes

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

The preparation of the Aroclor 1254 -induced rat S9 fraction was carried out according to MARON & AMES.

Test concentrations with justification for top dose:

Main study
Five concentrations ranging from 0.313 to 5.0 or 0.625 to 10.0 µg Amine, C10-16-alkyldimethyl, N-oxide (AO-1270)/mL were selected for the experiments without and with metabolic activation, respectively.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: aqua ad iniectibilia
- Justification for choice of solvent/vehicle: substance is soluble in water

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

aqua ad iniectabilia

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

aqua ad iniectabilia

True negative controls:

no

Positive controls:

yes

Positive control substance:

9,10-dimethylbenzanthracene

Remarks:

with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration:
Experiment 1: 4 hr with or without S9
Experiment 2: 4 hr with S9, 24 hr without S9
- Expression time (cells in growth medium): 8 days

SELECTION AGENT (mutation assays): 6-thioguanine

NUMBER OF REPLICATIONS: 5

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: relative plating efficiency

Evaluation criteria:

If in both independent experiments solvent and positive controls show results within the norm and if the test item does not increase the mutation frequency 2-fold above the mean of the solvent controls under any condition, or if the mutation frequency is always lower than 40 x 10-6 and if at least 1000000 cells per condition have been evaluated, the item is considered as negative in the test.

In case of a dose-dependent increase of the mutation frequency in both independent experiments (at similar concentrations) to at least 2-fold solvent control and at least 40 x 10-6 both in the presence and/or absence of S9 mix, the item is considered as positive in the test.

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

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

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none noted
- Effects of osmolality: none noted
- Evaporation from medium: not applicable
- Water solubility: completely soluble in water
- Precipitation: none noted
- Other confounding effects:

ADDITIONAL INFORMATION ON CYTOTOXICITY: Cytotoxicity in form of decreased plating efficiency (PE1) and (PE2) was noted in both experiments each carried out without and with metabolic activation at the top concentrations of 5.0 or 10.0 µg Amin, C10-16-alkyldimethyl, N-oxide (AO-1270)/mL medium, respectively.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Mutagenicity

Experiments without metabolic activation (4-h or 24-h exposure)

The mutation frequency of the negative controlaqua ad iniectabiliawas 6.67 and 5.10 x 10^-6 clonable cells. Hence, the negative controls were well within the expected range (see below).

The mutation frequency of the cultures treated with concentrations of 0.313, 0.625, 1.25, 2.5 or 5.0 µg Amin, C10-16-alkyldimethyl, N-oxide (AO-1270)/mL culture medium ranged from 1.21 to 7.10 x 10^-6 clonable cells. These results are within the normal range of the negative controls.

Experiments with metabolic activation (4-h exposure)

The mutation frequency of the negative controlaqua ad iniectabiliawas 5.94 and 2.05 x 10^-6clonable cells. Hence, the negative controls were well within the expected range (see below).

The mutation frequency of the cultures treated with concentrations of 0.625, 1.25, 2.5, 5.0 or 10.0 µg Amin, C10-16-alkyldimethyl, N-oxide (AO-1270)/mL culture medium ranged from 0.53 to 7.42 x 10^-6 clonable cells. These results are within the normal range of the negative controls.

The positive controls EMS (ethyl methanesulfonate) in the direct test and DMBA (9,10-dimethyl-1,2-benzanthracene) a compound which requires metabolic activation caused a pronounced increase in the mutation frequencies ranging from250.98to1038.82x 10^-6 clonable cells in the case of EMS and ranging from66.48to193.85x 10^-6 clonable cells in the case of DMBA, indicating the validity of this test system.

The background mutation frequency at LPT ranges from 1.30 to 38.36 x 10^-6clonable cells for the negative controls. The mutation frequency of the positive controls at LPT ranges from 112.1 to 1708.4 x 10^-6clonable cells forand 130.0 to 2693.3 x 10⁶clonable cells for DMBA.

No relevant change in pH value or osmolality was noted.

Tables of results are attached.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative