Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report date:
2000

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1997
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1,1,1,2,3,3-hexafluoro-2-(heptafluoropropoxy)-3-[(trifluorovinyl)oxy]propane
EC Number:
216-703-2
EC Name:
1,1,1,2,3,3-hexafluoro-2-(heptafluoropropoxy)-3-[(trifluorovinyl)oxy]propane
Cas Number:
1644-11-7
Molecular formula:
C8F16O2
IUPAC Name:
1,1,1,2,2,3,3-heptafluoro-3-({1,1,1,2,3,3-hexafluoro-3-[(1,2,2-trifluoroethenyl)oxy]propan-2-yl}oxy)propane
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch: 19B2059
- Expiration date of the lot/batch: 14 January, 2000
- Purity test date: 14 January, 2000

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Darkness at approximately 20C in a fume cupboard under inert (N2) conditions
- Stability under test conditions: Confirmed over 4 hours in deionized water
- Solubility and stability of the test substance in the solvent/vehicle: Confirmed over 4 hours in deionized water
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Emulsified in deionized water at appropriate concentrations immediately before use.

FORM AS APPLIED IN THE TEST: Emulsified in deionized water

Method

Target gene:
Histidien operon, tryptophan 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:
Aroclor 1254-induced rat liver homogenate S9-mix.
Test concentrations with justification for top dose:
50, 160, 500, 1600, 5000 ug/plate. Precipitation of the test article was visible at the 5000 ug/plate dose.
Vehicle / solvent:
Deionized water
Controls
Untreated negative controls:
yes
Remarks:
Untreated
Negative solvent / vehicle controls:
yes
Remarks:
Deionized water
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Preincubation period: Overnight (Approximately 12 hours)
- Exposure duration: 48 hours
- Expression time (cells in growth medium): approximately 60 hours
- Selection time (if incubation with a selection agent): 48 hours

SELECTION AGENT (mutation assays): Histidine or tryptophan-minimal agar

NUMBER OF REPLICATIONS: 2 independent runs (with three plates) of the test article and each control

NUMBER OF CELLS EVALUATED: All revertant colonies on each plate were counted.

DETERMINATION OF CYTOTOXICITY
- Method: Bacterial lawn thinning and a reduction in spontaneously occuring colonies.
Evaluation criteria:
A test article is classified as mutagenic if it has either of the following effects:
a) it produces at least a 2-fold increase in the mean number of revertants per plat 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.
b)it induces a dose-related 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 in at least two to three concentrations of the test compound at complete bacterial background lawn.

If the test substance does ont achieve either of the above criteria, it is considered to show no evidence of mutagenic activity in this system.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
An increased number of revertants was obtained in the absence of S9 mix, but this effect was not verified by a repeat experiment with finer graduated concentrations and therefore not considered relevant.
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
Based on the results of the study, the test article was not mutagenic in the Bacterial Reverse Mutation Assay in either the presence or absence of metabolic activation.
Executive summary:

The mutagenic potential of the test article was evaluated in the Bacterial Reverse Mutation Assay with Salmonella typhimurium (strains: TA100, TA1535, TA1537, and TA98) and Escherichia coli (strain: WP2uvrA) in the presence or absence of metabolic activation. This study was conducted in compliance with OECD GLP guidelines (1999). The study design was based on OECD 471 (1997), US EPA OPPTS 870.5100 (1998), EEC Directive 92/69 L383A Annex B13 and B14. The test article was emulsified in deionized water prior to administration. Two mutagenicity tests were performed with all strains (one plate incorporation test and one preincubation test) each in the presence or absence of metabolic activation (rat liver homogenate). Positive and negative controls were included in all tests. Each bacterial strain was exposed to the test article at up to 5000 ug/plate. A repeat preincubation test was performed with TA1537. The positive and negative controls performed as expected which indicated the tests were valid. A weak increase in the number of revertants was noted in the test article-treated group in strain TA1537 in the absence of metabolic activations. The test article did not increase the number of revertants in the repeat test with TA1537. No increases in the number of revertant colonies were noted in any other strain. Based on the results of the study, the test article was not mutagenic in the Bacterial Reverse Mutation Assay in either the presence or absence of metabolic activation.