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Toxicological information

Genetic toxicity: in vitro

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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:
6 July 2000- 19 July 2000 (experimental start and completion date)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted under GLP coditions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report Date:
2001

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Qualifier:
according to
Guideline:
other: Japan Ministry of Agriculture, Forestry and Fishereis (1985). Joint Directives of J EPA, J MHW and J MITI (31 October 1997). Kampoan No. 287, Eisei No. 127 and Kikyoku No. 2 (31 October 1997). Official Notice of J MOL (8 February 1999).
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report: T-7499; Perfluorobutanesulfonyl fluoride
- Substance type: Clear colorless liquid
- Physical state: Liquid
- Analytical purity: 96-98%
- Storage condition of test material: Room temperature

Method

Species / strain
Species / strain / cell type:
other: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and and E.coli WP2uvrA/pKM101 (CM891)
Additional strain / cell type characteristics:
other: The S. typhimurium strains contain histidine mutations and the E.coli strain contains a tryptophan dependant mutant (ochre mutation).
Metabolic activation:
with and without
Metabolic activation system:
S9 Fraction
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone. The control substances's solvent was DMSO. - Justification for choice of solvent/vehicle: the solubility of the test substance was assessed at 50 mg/ml in acetone, in which it was sufficently soluble.
Controls
Untreated negative controls:
yes
Remarks:
The solvent, acetone, was used as a negative control.
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Migrated to IUCLID6: and sodium azide, 2-Nitrofluorene, 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (in absence of S9-mix), 2-aminoantracene (in presence of S9 mix)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk
DURATION
- Preincubation period: 30 minutes

NUMBER OF CELLS EVALUATED: background lawn density of non-revertant cells were observed.
Evaluation criteria:
The mutagenicity of a test substance was assessed by applying the following criteria:A) if treatment with a test substance produces an incerase in revertant colony numbers of at least twice the concurrent solvent/vehiclle controls, with some evidence of positive dose relationship, in two separae experimens, with any bacterial strain either in the presence or absence of S9 mix, the test substance will be considered to show evididence of mutagenic activity in this test system. B) if treatment with a test substance does not produce reproducible increases of at least 1.5 times the concurrent solvent/vehicle controls in either mutation test, the test subtance will be considered to show no evidence of mutagenic activity in this test system. C) if the results obtained fail to satisfy the criteria for a clear "positive" or "negative" response given in parts A and B, even after the additional testing outlined in the mutation test procedure, the test data may be subjected to analysis to determine the statistical significance of any increases in revertant colony numbers.
Statistics:
No statisitics were used in parts A and B of the evaluation criteria described above. In part C, the statisitical procedures used will be thsoe decreibed by Mahon et al (1989) and will usually be analysis of variance followed by Dunnett's test. Biological signficance should always be considered along with statistical significant. It should be noted that it is acceptable to conclude an equivocal response if no clear results can be obtained.

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and E.coli WP2uvrA/pKM101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: No substantial increases in revertant colony numbers over control counts were obstained with any of the tester strains following exposure to the test article at any concentartion in either the presence or absence of S9 mix. Visible thinning of the background lawn of non-revertant cells, together with a reduction in revertant colony numbers, was obtained following exposure to the test article at 5000 micrograms/plate was selected for use in the second test.
COMPARISON WITH HISTORICAL CONTROL DATA: the test would be considered valid if the mean of the solvent/vehicle control revertant colony numbers for each strain should like within the 99% confidence limits of the current historical control range of the laboratory, unless otherwise justified by the study director.
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

The test article showed no evidence of mutagenic activity in the bacterial system under the test conditions employed.
Executive summary:

In the in vitro assessment of the mutagenic potential of T-7499, histidine dependent auxotrophic mutants of Salmonella typhimurium, strains TA135, TA 1537, TA98 and TA100, and a tryptophan dependent mutant of Escherichia coli, strain WP2uvrA/pKM101 (CM891), were exposed to the test article diluted in acetone. Acetone was also used as a negative control. Two independent mutation tests were performed in the presence and absence of liver preparations from Aroclor 1254-induced rats (S9 mix). In both tests, the tester strains were exposed to the test article in gas-tight glass vials during a preincubation stage prior to plating out. Concentrations of the test article up to 5000 micrograms per plate were tested in the mutation tests. This is the standard limit concentration recommended in the regulatory guidelines that this assay follows. Other concentrations used were a series of approximately half-log dilutions of the highest concentration. Toxicity (observed as thinning of the background lawn of non-revertant cells, together with a reduction in revertant colony numbers) was seen in all strains following exposure to the test article at 5000 micrograms per plate in both tests. No evidence of mutagenic activity was seen at any concentration the test article in either mutation test. The concurrent positive controls demonstrated the sensitivity of the assay and the metabolizing activity of the liver preparations. It is concluded that, under the test conditions employed, the test article showed no evidence of mutagenic activity in this bacterial system.