1-Propanaminium, 2-hydroxy-N,N,N-trimethyl-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)thio]-, chloride (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:

30 April, 2015 - 22 May, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(1997)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(2008)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

{2-hydroxy-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulfanyl]propyl}trimethylazanium chloride

EC Number:

811-523-6

Cas Number:

88992-45-4

Molecular formula:

C14 H19 F13 N O S . Cl

IUPAC Name:

{2-hydroxy-3-[(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)sulfanyl]propyl}trimethylazanium chloride

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): NS-4000
- Appearance: Off-white powder (determined by WIL Research Europe)
- Storage condition of test material: At room temperature desiccated

Method

Target gene:

- S. typhimurium: Histidine gene
- E. coli: Tryptophan gene

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9-mix induced by Aroclor 1254 (500 mg/kg).

Test concentrations with justification for top dose:

Experiment 1
Preliminary test (without and with S9) TA100 and WP2uvrA: 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate
Main study: TA1535, TA1537 and TA98:
Without and with S9-mix: 1.7, 5.4, 17, 52, 164 and 512 µg/plate
Experiment 2:
TA1535, TA1537, TA98 and TA100:
Without S9-mix: 1.7, 5.4, 17, 52, 164 and 512 µg/plate
With S9-mix: 5.4, 17, 52, 164, 512 and 1600 µg/plate
WP2uvrA:
Without and with S9-mix: 17, 52, 164, 512, 1600 and 5000 µg/plate

Vehicle / solvent:

- Vehicle used: Milli-Q
- Justification for choice of vehicle: Test compound was soluble in Milli-Q and Milli-Q has been accepted and approved by authorities and international guidelines. Test substance concentrations were used within 2.5 hours of preparation.

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Milli-Q

Positive controls:

yes

Positive control substance:

other: methylmethanesulfonate 650 µg/plate in DMSO for TA100

Remarks:

without S9

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Milli-Q

Positive controls:

yes

Positive control substance:

other: 2-nitrofluorene 10 µg/plate in DMSO for TA98

Remarks:

without S9

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Milli-Q

Positive controls:

yes

Positive control substance:

other: ICR-191 2.5 µg/plate in DMSO for TA1537

Remarks:

without S9

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Milli-Q

Positive controls:

yes

Positive control substance:

other: 4-nitroquinoline-N-oxide 10 µg/plate in DMSO for WP2uvrA

Remarks:

without S9

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Milli-Q

Positive controls:

yes

Positive control substance:

other: sodium azide 5 µg/plate in saline for TA1535

Remarks:

without S9

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Milli-Q

Positive controls:

yes

Remarks:

2-aminoanthracene in DMSO for all tester strains, with 1 μg/plate (TA98, TA100), 2 μg/plate (TA100), 2.5 μg/plate (TA1535, TA1537), 5 μg/plate (TA1539), 15 μg/plate (WP2unrA)

Remarks:

with S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hour

NUMBER OF REPLICATIONS:
- Doses of the test substance were tested in triplicate in each strain. Two independent experiments were conducted.

NUMBER OF CELLS EVALUATED: 10E8 per plate

DETERMINATION OF CYTOTOXICITY
- Method: The reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies.

OTHER EXAMINATIONS:
- The presence of precipitation of the test compound on the plates was determined.

Evaluation criteria:

A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537, TA98 or WP2uvrA is not greater than three (3) times the concurrent vehicle control.
b) The negative response should be reproducible in at least one follow-up experiment.

A test substance is considered positive (mutagenic) in the test if:
a) The total number of revertants in the tester strain TA100 is greater than two (2) times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537, TA98 or WP2uvrA is greater than three (3) times the concurrent vehicle control.
b) In case a follow up experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment.

Statistics:

Not performed.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

see additional information on results

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation was observed up to and including the top dose of 5000 µg/plate.

RANGE-FINDING/SCREENING STUDIES:
- In tester strain TA100, toxicity was observed at dose levels of 164 μg/plate and above in the absence of S9-mix and toxicity was observed at dose levels of 512 μg/plate and above in the presence of S9-mix. In tester strain WP2uvrA, toxicity was observed at the dose level of 5000 μg/plate in the absence and presence of S9-mix.

COMPARISON WITH HISTORICAL CONTROL DATA:
- The negative and strain-specific positive control values were within our laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Experiment 1:
TA1535: without S9: 512 µg/plate
TA1537: without S9: 512 µg/plate
TA98: without S9: 512 µg/plate
Experiment 2:
TA1535: without S9: 512 µg/plate and with S9: 512 µg/plate and above
TA1537 and TA98: without S9: 512 µg/plate and with S9: 1600 µg/plate
TA100: without S9: 164 µg/plate and above and with S9: 512 µg/plate and above
WP2uvrA: without and with S9: 5000 µg/plate

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

In an AMES test, performed according to OECD 471 guideline and GLP principles, NS-4000 was found not to be mutagenic up to and including 5000 μg/plate with or without metabolic activation.

Executive summary:

An AMES test was performed according to OECD 471 guideline and GLP principles. All bacterial strains showed negative responses up to and including cytotoxic concentrations in two experiments, i.e. no significant dose-related increase in the number of revertants with or without metabolic activation was seen. No precipitation of the test substance was observed. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Based on the results of this study it is concluded that NS-4000 is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay with or without metabolic activation.