N-[3-(dimethylamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctanesulphonamide 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:

experimental study

Adequacy of study:

key study

Study period:

06 January 2017 to 19 October 2017

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-[3-(dimethylamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctanesulphonamide N-oxide

EC Number:

279-481-6

EC Name:

N-[3-(dimethylamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctanesulphonamide N-oxide

Cas Number:

80475-32-7

Molecular formula:

C13H17F13N2O3S

IUPAC Name:

N-[3-(dimethylamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctanesulphonamide N-oxide

Test material form:

solid: particulate/powder

Details on test material:

Purity: 96.7 %

Method

Target gene:

The purpose of this study was to evaluate a test article for its ability to induce forward mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus (hprt) of Chinese hamster ovary (CHO) cells, as assayed by colony growth in the presence of 6-thioguanine (TG resistance, TGr).

Species / strain

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

- Properly maintained: yes- Periodically checked for Mycoplasma contamination: yes- Periodically checked for karyotype stability: yes- Periodically "cleansed" against high spontaneous background: yes

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9

Test concentrations with justification for top dose:

In the preliminary toxicity assay, the concentrations tested were 3.91, 7.81, 15.6, 31.3, 62.5, 125, 250, 500, 1000, and 2000 μg/mL. The maximum concentration evaluated was the limit dose for this assay.Due to lack of precipitation and cytotoxicity (10 to 20% adjusted relative survival), the concentrations chosen for the definitive mutagenicity assay were 125, 250, 500, 1000, and 2000 μg/mL with and without S9.Based on Sponsor request, the mutagenicity assay was repeated at concentrations of 125, 250, 500, 1000, and 2000 μg/mL with S9.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO to dilute positive controls, Ethanol: Water (1.2:1) for the test article- Justification for choice of solvent/vehicle:Ethanol: Water (1.2:1) was the vehicle of choice based on thesolubility of the test article and compatibility with the target cells.

Controls

Untreated negative controls:

yes

Remarks:

Untreated controls were included in the preliminary cytotoxicity assay in order to compare the cytotoxicity and/or mutagenicity data with that of the vehicle control.

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

ethylmethanesulphonate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium
DURATION- Exposure duration: 5 +/- 0.5 hours- Expression time (cells in growth medium): 7 to 8 days- Selection time (if incubation with a selection agent): 7 to 10 days
SELECTION AGENT (mutation assays): 6-thioguanine
NUMBER OF REPLICATIONS: single cultures in the preliminary toxicity assay; duplicate cultures in the mutagenicity assays
NUMBER OF CELLS EVALUATED: 2.4 x 10E6 cells per culture
DETERMINATION OF CYTOTOXICITY- Method: adjusted relative survival

Evaluation criteria:

The test substance will be considered to have produced a positive response if it induces a dose-related increase in mutation frequency and an increase exceeding 95% historical vehicle control limits in at least one test dose level(s) as compared with concurrent vehicle control (p<0.01). If only one criterion is met (a statistically significant or dose-dependent increase or an increase exceeding the historical control 95 % confidence interval), the result will be considered equivocal. If none of these criteria are met, the results will be considered to be negative. Other criteria also may be used in reaching a conclusion about the study results (e.g., comparison to historical control values, biological significance, etc.). In such cases, the Study Director will use sound scientific judgment and clearly report and describe any such considerations.

Statistics:

Statistical analyses were performed using the method of Snee and Irr (1981), with significance established at the 0.05 level.

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Preliminary toxicity: Adjusted relative survival (ARS): 86.42% and 56.33% at 2000 μg/mL + and - S9 , respectively Mutagenicity: average ARS: 112.1 and 98.52% at 2000 μg/mL + and - S9 , respectively Retest: average ARS: 95.02% at 2000 μg/mL + S9

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: The test article did not have an adverse impact on the pH of the cultures (pH 7.5 at the top dose).
- Effects of osmolality: Preliminary Toxicity Assay: The osmolality of the cultures was acceptable as it did not exceed the osmolality of the vehicle control by more than 120%.
- Precipitation: No visible precipitate at the beginning or end of treatment

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Under the conditions of the assay described in this report, 1-Octanesulfonamide, N-[3-(dimethyloxidoamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluoro- was concluded to be negative for the induction of forward mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus (hprt) of Chinese hamster ovary (CHO) cells, in the presence and absence of an exogenous metabolic activation system, in the in vitro mammalian cell forward gene mutation (CHO/HPRT) assay.

Executive summary:

The test article, 1-Octanesulfonamide, N-[3-(dimethyloxidoamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluoro-, was evaluated for its ability to induce forward mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus (hprt) of Chinese hamster ovary (CHO) cells, in the presence and absence of an exogenous metabolic activation system, as assayed by colony growth in the presence of 6-thioguanine (TG resistance, TG^r). Ethanol : Water (1.2:1) was used as the vehicle.

In the preliminary toxicity assay, the concentrations tested were 3.91, 7.81, 15.6, 31.3, 62.5, 125, 250, 500, 1000, and 2000 µg/mL. The maximum concentration evaluated was the limit dose for this assay. No visible precipitate was observed at the beginning or end of treatment. Due to lack of precipitation and cytotoxicity (10 to 20 % adjusted relative survival), the concentrations chosen for the definitive mutagenicity assay were 125, 250, 500, 1000, and 2000 µg/mL with and without S9.

In the initial mutagenicity assay, no visible precipitate was observed at the beginning or end of treatment. Due to lack of precipitation and cytotoxicity (10 to 20 % adjusted relative survival), cultures treated at all concentrations were chosen for mutant selection. No significant increases in mutant frequency, as compared to the concurrent vehicle controls (p>0.01), were observed at any concentration evaluated with or without S9.   The positive controls induced significant increases in mutant frequency (p < 0.01).

Based on Sponsor request, the mutagenicity assay was repeated at concentrations of 125, 250, 500, 1000, and 2000 µg/mL with S9. In the repeat assay, no visible precipitate was observed at the beginning or end of treatment. Due to lack of precipitation and cytotoxicity (10 to 20 % adjusted relative survival ), cultures treated at all concentrations were chosen for mutant selection. No significant increases in mutant frequency, as compared to the concurrent vehicle controls (p>0.01), were observed at any concentration evaluated with S9.   The positive control induced significant increases in mutant frequency (p < 0.01).

These results indicate 1-Octanesulfonamide, N-[3-(dimethyloxidoamino)propyl]-3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluoro- was negative for the ability toinduce forward mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus (hprt) of Chinese hamster ovary (CHO) cells, in the presence and absence of an exogenous metabolic activation system.