Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was selected as a key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
Qualifier:
according to guideline
Guideline:
other: EC Commission Directive 2000/32/EC, Annex 4D-B13/14 No. L136
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl methacrylate
EC Number:
218-407-9
EC Name:
3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl methacrylate
Cas Number:
2144-53-8
Molecular formula:
C12H9F13O2
IUPAC Name:
3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-methylprop-2-enoate
Details on test material:
- Purity: 97.59%

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced rat liver S9
Test concentrations with justification for top dose:
33.3, 66.7, 100, 333, 667, 1000, 3333, and 5000 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Ethanol
- Justification for choice of solvent/vehicle: Based on the solubility of the test substance and compatibility with the target cells.
Controls
Untreated negative controls:
yes
Remarks:
ethanol
Negative solvent / vehicle controls:
yes
Remarks:
ethanol
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (TA1535, TA1537, TA100, WP2 uvrA +S9), benzo[a]pyrene (TA98 +S9), 2-nitrofluorene (TA98 -S9), sodium azide (TA100 and TA1535 -S9), Acridine mutagen ICR-191 (TA1537 –S9), 4-nitroquinoline-N-oxide (WP2 urvA -S9)
Remarks:
All positive controls were diluted with dimethyl sulfoxide (DMSO) except sodium azide and ICR-191, which were diluted in sterile water.
Details on test system and experimental conditions:
METHOD OF APPLICATION: In the non-activated assays, 0.5 mL of sham mix and 100 μL of vehicle, test substance dilution, or positive control were added to pre-heated (45–48°C) glass culture tubes containing 2 mL of selective top agar, followed by 100 μL of tester strain. In the S9-activated assays, 100 μL of the vehicle, test substance dilution, or positive control were added to pre-heated (45–48°C) glass culture tubes containing 2 mL of selective top agar, followed by 100 μL of tester strain and 0.5 mL of S9 mix. All mixtures were vortexed and overlaid onto the surface of minimum glucose agar plate.

DURATION
- Preincubation period: none
- Exposure duration: approximately 48-52 hours at 37 ± 2°C
- Expression time (cells in growth medium): approximately 48-52 hours. Plates that were not evaluated immediately following incubation were stored at approximately 4°C.

NUMBER OF REPLICATIONS: toxicity-mutation tests were plated in duplicate; mutagenicity tests plated in triplicate

NUMBER OF CELLS EVALUATED: tester stain culture density was approximately 10e9 cells per mL.

DETERMINATION OF CYTOTOXICITY
A minimum of 3 non-toxic scorable dose levels were required to validate the study. A dose level was considered toxic if it caused:
- A >50% reduction in the mean number of revertants per plate relative to the mean negative control value and exhibited a dose-dependent drop in the revertant count, or
- A reduction in the background lawn.
Evaluation criteria:
For the test substance to be evaluated positive, it must cause a dose-related increase in the mean revertants per plate unless observed at the top dose level only. Data sets for tester strains TA1535 and TA1537 were judged positive if the increase in mean revertants at the highest numerical dose response was ≥ 3.0-fold the mean concurrent negative control value (vehicle control). Data sets for tester strains TA98, TA100 and WP2 uvrA were judged positive if the increase in mean revertants at the highest numerical dose response was ≥ 2.0-fold the mean concurrent negative control value (vehicle control).
Statistics:
For each selected tester strain, the mean number of revertants and the standard deviation at each concentration with and without S9 activation were calculated.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and 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
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
Additional information on results:
RANGE-FINDING/SCREENING STUDIES:
No positive mutagenic responses were observed at any dose level in any tester strain in the absence or presence of S9 metabolic activation. No toxicity was observed at any dose level with any tester strain in either the absence or presence of S9. Test substance precipitation was observed starting at 667 μg/plate in both the activated and non-activated test system.

MUTAGENICITY TEST:
No toxicity was observed at any dose level with any tester strain in either the absence or presence of S9. Test substance precipitation was observed starting at 667 μg/plate for TA98, TA100, TA1535, and TA1537 in presence of S9 activation and for WP2uvrA and TA98 in the absence of S9 activation. Test substance precipitation was observed starting at 1000 μg/plate for TA100 in the absence of S9 activation and for WP2uvrA in the presence of S9 activation. For TA1535 and TA1537 in the absence of S9 activation test substance precipitation was observed starting at 3333 μg/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 in the absence and presence of S9 activation

The study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).
Negative when tested in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and in Escherichia coli WP2 uvr A in the absence and presence of S9 activation.
Executive summary:

The test substance was evaluated for mutagenicity in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and in Escherichia coli WP2uvrA with and without an exogenous metabolic activation system (Aroclor-induced rat liver S9). The maximum concentration tested was 5000 µg/plate. No evidence of mutagenic activity was detected in either the absence or presence of Aroclor-induced rat liver S9. In this study, the test substance was negative.