2,2,3,3,4,4,5,5,6,6-decafluoro-6-trifluorovinyloxyhexanenitrile

Genetic toxicity: in vitro

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

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:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

Version / remarks:

1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2,3,3,4,4,5,5,6,6-decafluoro-6-trifluorovinyloxyhexanenitrile

EC Number:

443-050-2

EC Name:

2,2,3,3,4,4,5,5,6,6-decafluoro-6-trifluorovinyloxyhexanenitrile

Cas Number:

120903-40-4

Molecular formula:

C8F13NO

IUPAC Name:

2,2,3,3,4,4,5,5,6,6-decafluoro-6-[(1,2,2-trifluoroethenyl)oxy]hexanenitrile

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: 3M, Lot 3
- Purity, including information on contaminants, isomers, etc.: 93.8%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: approximately 5°C in a refrigerator, light protected, under nitrogen.
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: no data.
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: no data.
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: confirmed over 4 hours in ethanol.
- Reactivity of the test material with the incubation material used (e.g. plastic ware): no data.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): dissolved in ethanol at appropriate concentrations immediately before use.
- Preliminary purification step (if any): no data.
- Final concentration of a dissolved solid, stock liquid or gel: concentrations of: 50, 160, 500, 1600, and 5000 μg/plate.
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): no data.

FORM AS APPLIED IN THE TEST (if different from that of starting material)
- Specify the relevant form characteristics if different from those in the starting material, such as state of aggregation, shape of particles or particle size distribution: no data.

Method

Target gene:

Histidine and tryptophan operons.

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system:
Aroclor-induzierter Rattenleber S9-Mix "english" Aroclor-induced rat-liver S9-mix

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 50, 160, 500, 1600, and 5000 ug/plate.
Concentration range in the main test (without metabolic activation): 50, 160, 500, 1600, and 5000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-aminoanthracene for all strains with S9

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration (single, duplicate, triplicate) : triplicate
- Number of independent experiments : 2

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): No data.
- Test substance added in agar (plate incorporation)

TREATMENT AND HARVEST SCHEDULE:
- Preincubation period, if applicable: no data.
- Exposure duration/duration of treatment: 48 hours.

FOR GENE MUTATION:
- Expression time (cells in growth medium between treatment and selection): 48 hours
- Selection time (if incubation with a selective agent): 48 hours

Rationale for test conditions:

Per OECD 471.

Evaluation criteria:

The assay is considered valid if the following criteria are met:
- The solvent control data are within the lab's normal control range for spontaneous mutant frequency
- The positive controls induce increases in the mutation frequency which are significant and within the laboratory's normal range.

Criteria for a positive response:
A test criteria is considered mutagenic if it has either of the following effects:
- It produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial background lawn.
- It produces a dose-related increase in the mean number of revertants per plate in at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.
If the test substance does not achieve either of the above criteria, it is considered to show no mutagenic activity in this system.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Data on pH: No data.
- Data on osmolality: No data.
- Possibility of evaporation from medium: Not expected based on vapor pressure and boiling point.

Ames test:
- Signs of toxicity : none observed.
- Individual plate counts : counted following 48 hour exposure.
- Mean number of revertant colonies per plate and standard deviation : revertant colonies were comparable to solvent controls at all exposure level. Standard deviation were within historical control ranges.

Applicant's summary and conclusion

Conclusions:

Based on the results of the test, MV5CN is not mutagenic in an Ames assay with and without metabolic activation (S9).

Executive summary:

The mutagenic potential of MV5CN was evaluated in the Bacterial Reverse Mutation Assay with S. typhimurium strains TA98, TA100, TA 102, TA1535 and TA1537 in both the presence and absence of a metabolic activation system (S9-mix: Aroclor 1254-induced rat liver). This study was performed in compliance with OECD GLP (1999). The test method was based on OECD 471 (1997), U.S. EPA OPPTS 870.5100 (1998), and EC Directive 2000/32/EC, L136, Annex 4D, Part B 13/14. Two independent mutagenicity studies were conducted (first plate incorporation test with 10% and second plate incorporation test with 30% S9-mix), each in the absence and presence of a metabolizing system. For both studies, MV5CN was dissolved in ethanol. The concentrations used for the first plate incorporation test (10% S9-mix) were 50, 160, 500, 1600, and 5000 ug per plate. The test compound did not precipitate on the plates up to the highest investigated dose of 5000 ug/plate. Because of toxicity in the first plate incorporation test, dose levels from 5 to 5000 ug/plate were chosen for the second plate incorporation test (30% S9-mix). Strain-specific positive controls were included in all tests. All criteria for a valid study were met as described in the protocol. No increase in revertant colonies was observed in either the presence or absence of S9-mix for any strain at any concentration. Under the conditions of this study, MV5CN was not mutagenic in the Bacterial Reverse Mutation Assay in either the presence or absence of metabolic activation.