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Toxicological information

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:
17 October 2001-13 March 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted under GLP conditions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report Date:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
other: EEC Directive 67/548/EEC, part B (2000)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report:T-7600
- Physical state: solid
- Composition of test material, percentage of components: >97% 2-Propenoic acid, 2-[Methyl[(nonafluorobutyl)sulfonyl]amino]ethyl ester, <2% water, < 0.01% Phenothiazine
- Lot/batch no.: Lot 1
- Expiration date of the lot/batch:06 August 2002
- Stability under test conditions: Not indicated
- Storage condition of test material: At room temperature in the dark

Method

Target gene:
detects base-pair and frameshift mutations
Species / strain
Species / strain / cell type:
other: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and Escherichia coli WP2uvrA
Details on mammalian cell type (if applicable):
- Properly maintained: Yes, Salmonella strains were regulatry checked to confirm histidine-requirement, crystal violet sensitivity, ampicillin resistance (TA98 and TA100), UV-sensitivity and the number of spontaneous revertants. The E.coli strains were regulatory checked to confirm teh tryptophan-requirement, UV-sensitivity and the number of spontaneous revertants.
- Periodically checked for Mycoplasma contamination: No
- Periodically checked for karyotype stability: No
- Periodically 'cleansed' against high spontaneous background:No
Additional strain / cell type characteristics:
other: Th E.coli strain lack an excision repair system.
Metabolic activation:
with and without
Metabolic activation system:
S9-mix (5 and 10%)
Test concentrations with justification for top dose:
3, 10, 33, 100, 333, 1000, 3330 micrograms/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used:DMSO
- Justification for choice of solvent/vehicle: used as a known negative control.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
Saline and DMSO were used for reference.
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Migrated to IUCLID6: positive control for strain TA1535
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: no data
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours
NUMBER OF REPLICATIONS: Three
Evaluation criteria:
This mutation assay is considered acceptable if if meets A) the negative control data is within the laboratory background historical range for each tester strain. B) The positive control chemcials should produce responses in all tester strains which are within the laboratoy historical range documents for each positive control substance 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.
Statistics:
A test substance is considered negative if a) the total number of revertants in any tester strain at any concentration is not greater than two times the solvent control value, with or without metabolic activiation. b) The negative response should be reproducible in at least one independently repeated experiment. A test substance is considered postive if a) it induces a number of revertant colonies, dose related, greater than two-times the number of vertants induced by the solvent control in any of the test strains, either with or without metabolic activation. However, any mean plate count of less than 20 is considered to be not significant. b) the positive response should be reproducible in at least one independently repeated experiments.

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and and E.coli WP2uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
other: all controls were within the laboratory background historical control data ranges indicating that the metabolic activation system functioned properly.
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: In the dose range finding test, the test article was tested up to concentrations of 5000 micrgrams/plate with and without S9-mix in the trains TA100 and WP2uvrA. The test article precipiated on the plates at dose levels of 3330 and 5000 micrograms/plate in the presence of S9-mix. In tester strain Wp2uvrA, no toxicity was observed at all concentrations tested.
COMPARISON WITH HISTORICAL CONTROL DATA: All bacterial strains showed negative responses over the entire dose range, (i.e. no dose-related, two-fold, increase in the number of revertants in two independently repeated experiments.)
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

The results of this study indicated thatt the test article is not mutgenic in the Salmonella typhimurium reverse mutation asasy and in the Escherichia coli reverse mutation assay.
Executive summary:

The test article, C4 -Acrylate, was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of S.typhi TA1535, TA1537, TA100 and TA96) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli Wp2uvrA. The test was performed in two independent experiments in the presence and absence of S9-mix. In the dose range finding test, the test article was tested up to concentrations of 5000 micrograms/plate with and without S9-mix in the trains TA100 and WP2uvrA. The test article precipitated on the plates at dose levels of 3330 and 5000 micrograms/plate in the presence of S9-mix. In tester strain Wp2uvrA, no toxicity was observed at all concentrations tested. In the first and in the second mutation assay, the test article was tested up to concentrations of 3330 micrograms/plate in the absence and presence of S9-mix. The test article precipitated on the plates at this dose level. Toxicity was observed in test strains TA1537 and TA100 in the presence of S9-mix in the second mutation assay at the highest concentration tested. In all other strains tested no toxicity was observed at any of the concentrations tested. The presence of 5 and 10% (v/v) S9 activation did not influence these findings. The test article did not induce a dose related, two-fold increase in the number of revertant Histidine+ colonies in each of the four S.typhi test strains and in the number of revertant (trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation.

These results were confirmed in an independently repeated experiment. Based on the results of these studies, the test article is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.