Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot 2/C4F9S03-K
- Expiration date of the lot/batch: 04-06-01
- Purity test date: 01/17/02

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Solubility and stability of the test substance in the solvent/vehicle: The maximum solubility of test article in DMSO was 300 mg/mL.

Method

Target gene:
Histidine operon (S. typhimurium) and tryptophan operon (E. coli)
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Details on mammalian cell lines (if applicable):
CELLS USED
- Source of cells: Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 were originally obtained from Dr. Bruce N. Ames, University of California, Berkley. Escherichia coli strain WP2uvrA was obtained from Ms. Judy Mayo of Pharmacia and Upjohn Co., Kalamazoo, Michigan.
Metabolic activation:
with and without
Metabolic activation system:
Exogenous metabolic activation system consisting of Aroclor 1254 or phenobarbital-induced rat liver S-9 in 0.15M KC1 plus cofactors (S-9 mix). The components of the standard S-9 mix prepared from Aroclor 1254 or phenobarbital-induced, Sprague-Dawley rats.
Test concentrations with justification for top dose:
50, 100, 500, 1000 and 5000 ug/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Based on the results of a solubility test, DMSO was used as the solvent for this test article.
Controls
Negative controls:
yes
Solvent controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
methylmethanesulfonate
other: 2-Aminoanthracene
Remarks:
Positive chemicals used for the tester strains in the presence and absence of exogenous metabolis activation. All positive control substances were dissolved in DMSO.
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation) and preincubation
- Cell density at seeding (if applicable): 5 x 10^8 to 1 x 10^9 cells/mL

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48-72 hours
- Expression time (cells in growth medium): Plates were incubated at 37C for appxoximately 20 hours
Evaluation criteria:
A response was considered negative if all the strains treated with the test article had mean reversion frequencies that were less than twice that of the mean reversion frequencies of the corresponding solvent control plates in TA 98 and TA 100 and less than three times in TA 1535, TA 1537 and WP2uvrA, and there was no evidence of a dose-dependent response.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Positive controls valid:
yes
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Positive controls valid:
yes

Applicant's summary and conclusion

Conclusions:
Based on the results of the study, potassium perfluorobutane sulfonate did not result in genotoxicity to Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 and Escherichia coli strain WP2uvrA in the presence or absence of exogenous metabolic activation at a dose of 50 -5000 ug/plate.
Executive summary:

The mutagenic potential of potassium perfluorobutane sulfonate was evaluated in the Bacterial Reverse Mutation Assay with Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 and Escherichia coli strain WP2uvrA in the presence and absence of metabolic activation (S9 mix; Aroclor 1254 -induced rat liver). The study was performed using potassium perfluororbutane sulfonate dissolved in DMSO at concentrations of 50, 100, 500, 1000 and 5000 ug/plate based on a range finding study. The first definitive mutation assay (B-1), using the plate incorporation method, was performed with the four S. typhimurium strains and with E. coli WP2uvrA. Treatment during B-1 was performed by adding either 500 uL deionized, distilled water or 500 uL of rat S-9 cofactor mix to tubes containing 2.0 mL top agar supplemented with 1X histidine-biotin or 1X tryptophan solution. Immediately after, 100 uL of strains TA 98, TA 100, TA 1535, TA 1537 or WP2uvrA were added followed by the appropriate test article dose or solvent. Positive controls were treated with 100 uL of the appropriate stock solution. Tubes were vortexed for 2 -3 seconds after and contents were evenly distributed over a Vogel-Bonner bottom agar plate. Plates were solified on a level surface, inverted, and then incubated at 37C for approximately 70.5 hours. Plates, starting with the highest test article concentration, were observed for the presence of precipitate. Plates with no precipitate were counted using an automatic colony counter (ARTEK Counter, Model 880) and those with precipitate were counted by hand. Three counts were taken by rotating the plate on the counter stage and the mediun count was entered into a validaed, Lotus 123 spreadsheet program. The background lawn was also evaluated. Based on the results of the study, potassium perfluorobutane sulfonate did not result in genotoxicity to Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 and Escherichia coli strain WP2uvrA in the presence or absence of exogenous metabolic activation at a dose of 50 -5000 ug/plate.