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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:
27 May - 06 Jun 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Health Care Inspectorate, Ministry of Health, Welfare and Sport, Den Haag, The Netherlands
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 2-chloroacrylate
EC Number:
801-260-5
Cas Number:
96383-55-0
Molecular formula:
C11H6ClF13O2
IUPAC Name:
3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-chloroacrylate
Details on test material:
- Name of test material (as cited in study report): C6SFCLA
- Physical state: clear colourless liquid
- Lot/batch No.: G2X01
- Expiration date of the lot/batch: 31 Oct 2014
- Storage condition of test material: At room temperature in the dark

Method

Target gene:
"his operon" (for S. typhimurium strains) and "trp operon" (for E. coli strains).
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:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Phenobarbital and Naphthoflavone
Test concentrations with justification for top dose:
Range-finding assay: 3, 10, 33, 100, 333, 1000, 3330, 5000 µg/plate
Experiment 1 (with and without S9-mix): 100, 333, 1000, 3330, 5000 µg/plate; (results from the range finding assay for TA 100 and WP2uvrA are included in the results for experiment 1)
Experiment 2 (with and without S9-mix): 100, 333, 1000, 3330, 5000 µg/plate
Vehicle / solvent:
- Vehicle/solvent used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
2-nitrofluorene
sodium azide
methylmethanesulfonate
other: Without S9-mix: ICR-191 (TA 1537, 2.5 µg); sodium azide (TA 1535, 5 µg); 2-nitrofluorene (TA98, 10 µg); MMS (TA100, 650 µg); 4-NQO (WP2uvrA, 10 µg); With S9-mix: 2-aminoanthracene (TA1535, 2.5/5 µg; TA1535, 2.5 µg; TA98,1 µg;TA100,1/2.5 µg; WP2uvrA,10 µg)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48±4 h

NUMBER OF REPLICATIONS: triplicates each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: bacterial background lawn, number of revertants

METABOLIC ACTIVATION SYSTEM:
- Every batch of S9-mix was characterised with Benzo-(a)-pyrene and 2-aminoanthracene before use.
Evaluation criteria:
Acceptability criteria:
a) The negative control data (number of spontaneous revertants per plate) should be within the laboratory historical range for each tester strain.
b) The positive control chemicals should produce responses in all tester strains, which are within the laboratory historical range documented for each positive control substance. Furthermore, the mean plate count should be at least three times the concurrent vehicle control group mean.
c) The selected dose range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.

A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537, TA98 or WP2uvrA is not greater than three (3) times the concurrent vehicle control.
b) The negative response should be reproducible in at least one independently repeated experiment.

A test substance is considered positive (mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 is greater than two (2) times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537, TA98 or WP2uvrA is greater than three (3) times the concurrent vehicle control.
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one independently repeated experiment.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
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 applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: not observed up to the limit concentration

RANGE-FINDING/SCREENING STUDIES:
No reduction of the bacterial background lawn and no biologically relevant decrease in the number of revertants were observed.
In the dose range finding test, no increase in the number of revertants was observed upon treatment with C6SFCLA under all conditions tested.

COMPARISON WITH HISTORICAL CONTROL DATA:
In strain TA1535 (first experiment, absence of S9-mix), a fluctuation in the number of revertant colonies above the laboratory historical control data range was observed at the dose level of 5000 μg/plate. However, since the increase was not three-fold (a maximum of 1.3-fold was reached), this increase was not considered to be relevant.
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

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