1,1,2,2,3,3,4,4,4-nonafluoro-N-(2-hydroxyethyl)-N-methylbutane-1-sulphonamide

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:

16 October 2001 to 09 Novermber 2001 (dates of study plan)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2001

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: European Economic Community Directive 67/548/EEC, Part B.

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

T-7599

IUPAC Name:

T-7599

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): T-7599
- Substance type: Mono-constituent substance
- Physical state: Solid
- Analytical purity: >99.9% 1-Butanesulfonamide,1,1,2,2,3,3,4,4,4-Nonafluoro-n-(2-Hydroxyethyl)-N-Methyl-; < 0.1% Methylamine
- Lot/batch no.: Lot 6
- Expiration date of the lot/batch: 07 August 2002
- Storage condition of test material: at room temperature in the dark

Method

Target gene:

Histidine operon

Species / strain

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:

S9-fraction

Test concentrations with justification for top dose:

The test article was tested at concentrations of 3, 10, 33, 100, 333, 1000, 3330, and 5000 ug/plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Saline and DMSO
- Justification for choice of solvent/vehicle: Physiological saline used for TA1535, TA1537 and TA98 strains without metabolic activation. DMSO was used as a solvent for the remaining strains. DMSO was used as a negative control because it was used as a vehicle.

Controls

Untreated negative controls:

yes

Remarks:

Number of spontaneous revertants per plate

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: W/o S9: TA 1535- sodiumm azide; TA 1537- 9-aminoacridine ; TA98- daunomycine; TA100- methylmethanesulfonate; WP2uvrA- 4-nitroquinoline N-oxide. W S9: TA 1537, TA 1535, TA98, TA100, WP2uvrA- 2-aminoanthracene.

Details on test system and experimental conditions:

Each tester strain had the following additional mutations: Defective lipopolysaccharide cellcoat, mutation in the galactose metabolism; mutation in nitrate reductase; defective biotin synthesis, loss of the excision repair system-deletion of the ultraviolet-repair B gene.

METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours
OTHER EXAMINATIONS:
- Other: Colony counting

Evaluation criteria:

The test substance is considered negative in the test if the total number of revertants in any tester strain at any concentration is less than two times the solvent control value. The negative response should be reproducible in at least one independently repeated experiment. A test substance is considered positive if it induces a number of revertant colonies greater than two-times the number of revertants induced by the solvent control in any of the tester strains with or without metabolic activation. The positive response should be reproducible in at least one independently repeated experiment.

Results and discussion

Test results

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

Remarks:

The test article was cyctotoxic at 5000 ug/plate in TA 1535 and TA1537 without S9 mix and in the same strains at 3300 and 5000 ug/plate with S9 mix.

Vehicle controls validity:

other: The vehicle for the test article (DMSO) was used as a negative control.

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: The test substance did not precipitate in the top agar. Precipitation of the test article o the plates was not observed at the start or at the end of the incubation period in tester strain TA100 and WP2uvrA.
RANGE-FINDING/SCREENING STUDIES: The test article was tested in strains TA 100 and Wp2urA with concentrations of 3, 10, 33, 200, 333, 1000, 3330 and 5000 micrograms per plate in the absence and presence of 5% (v/v) S9-mix. The test article did not precipitate in the top agar and was not observed at the start or at the end of the incubation period in tester strains. No reduction of the bacterial background lawn and no biologically relevant decrease in the number of revertants was observed. No increase in the number of revertants was observed upton treatment with T-7599 under all conditions tested. The highest concetration of the test article used in the subsequent mutation assay was the recommended 5 mg/plate or thelevel at which the test substance inhibited baceterial growth.
COMPARISON WITH HISTORICAL CONTROL DATA: The reverse mutation assays are considered acceptable if the negative control data should be within the laboratory background hisorical range for each tester strain, the positive control chemcials should produce responses in all tester strains which are within the laboratory historical range documented for each positive control substance and the selected dose range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 6 mg/plate.

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

All bacterial strains showed negative responses over the entire dose range (no dose-related, two-fold, increase in the number of reverants in two independently repeated experiments). The negative and strain-specific positive control values were within the laboratoy background historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Based on the results of the study, it is concluded that the test article is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Executive summary:

The test article was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (IA1535, TA1537, TA100 and TA98) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli WP2uvrA. The test was performed in two independent experiments in the presence and absence of S9-mix. In the dose range finding test, the test article was tested up to concentrations of 5000 and 4740 micrograms/plate in the strains TA100 and WP2uvrA, respectively in the absence and presence of 5% (v/v) S9-mix. The test article did not precipitate on the plates at this dose level. In tester strain TA100, toxicity was observed at dose levels of 3330 and 5000 micrograms/plate in the absence of S9-mix and at a dose level of 1000 micrograms/plate and upwards already in the presence of S9-mix. In tester strain WP2uvrA, no toxicity was observed at any of the dose levels tested. In the first mutation assay, the test article was tested up to concentrations of 5000 micrograms/plate in the absence and presence of 5% (v/v) S9-mix in the strains TA1535, TA1537 and TA98. Toxicity was observed in all three tester strains. In the second mutation assay, the test article was tested up to concentrations of 3330 micrograms/plate in tester strain TA1535, TA1537 and TA100 and up to 5000 micrograms/plate in tester strain TA98 and WP2uvrA in the absence and presence of 10% (v/v) S(-mix. Toxicity was observed in all tester strains, except in test strain WP2uvrA in the presence of S9-mix. The test article did not induce a dose-related, two-fold increase in the number of revertant (His+) colonies in each of the four tester strains and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in an independently repeated experiment. Based on the results of this study, it is concluded that the test article is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.