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
Study period:
06 February 2007 to 15 February 2007
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:
2007
Report date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
In the range finding test, the S9-fraction was 5.6% (v/v). Amount of the S9-fraction in the S9-mix was increased by 12% and positive and negative control data were within historical range for both tester strains.Deviation has no influence on test result
Qualifier:
according to guideline
Guideline:
other: European Economic Community (EEC), Directive 2000/32/EC, Part B: Methods for the Determination of Toxicity; B. 13/14: "Mutagenicity: "Reverse Mutation Test using bacteria". EEC Publication Commission Directive (Published June 8, 2000).
Deviations:
yes
Remarks:
In the range finding test, the S9-fraction was 5.6% (v/v). Amount of the S9-fraction in the S9-mix was increased by 12% and positive and negative control data were within historical range for both tester strains.Deviation has no influence on test result
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
- Name of test material (as cited in study report): MTDID 6675
- Substance type: Clear colourless liquid
- Physical state: Liquid
- Analytical purity: Formulation tested is 30.0% active ingredient in water
- Purity test date: 12/20/2005
- Lot/batch no.: 140499-8/25
- Expiration date of the lot/batch:02 July 2008
- Storage condition of test material: Refrigerated (2-8 degrees C) in the dark

Method

Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA1535, TA1537, TA98, TA100, and Escherichia coli WP2uvrA.
Additional strain / cell type characteristics:
other: rfa: deep rough (defective lipopolysaccharide cellcoat) gal: mutation in the galactose metabolism chl: mutation in nitrate reductase bio: defective biotin synthesis uvrB: loss of the excision repair system (deletion of the ultraviolet-repair B gene)
Metabolic activation:
with and without
Metabolic activation system:
Rat liver microsomal enzymes were routinely prepared from adult male Wistar rats, which were obtained from Charles River, Sulzfel, Germany.
Test concentrations with justification for top dose:
100-5000 micrograms test article per plate.
Vehicle / solvent:
Vehicle(s)/solvent(s) used: None (test article was supplied as 30% active ingredient in water).
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: Without Metabolic Activation: TA1535: Sodium azide, TA1537: 9-aminoacridine, TA98: 2-nitrofluorene, TA100: methylemthanesulfonate, WP2uvrA: 4-nitroquinoline N-oxide. With Metabolic Activation: All Strains: 2-aminoanthracene.
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 hours of incubation time.
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Results and discussion

Test results
Species / strain:
other: Salmonella typhimurium TA1535, TA1537, TA98, TA100, and Escherichia coli WP2uvrA.
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
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

Based on the results of this study, the test article is not mutagenic in the Salmonella typhimurium reverse mutation assay and the Escherichia coli reverse mutation assay.
Executive summary:

The mutagenic activity of the test article (liquid, batch 140499-19/10) was evaluated in the Salmonella typhimurium reverse mutation assay (strains: TA1535, TA1537, TA98 and TA100, histidine-requiring) and the Escherichia coli reverse mutation assay (strain: WP2uvrA, tryptophan-requiring) in the presence and absence of a metabolic activation system (S9-mix: rat liver S9-mix induced by a combination of phenobarbital and ß naphthoflavone) following OECD guideline No. 471 “Genetic Toxicology: Bacterial Reverse Mutation Test” (adopted July 21, 1997). The test material was diluted in Milli-Q water resulting in a clear colorless liquid that was filter (22 µm) sterilized and used within 3 hours of preparation. A conversion factor for purity (29.9 ± 0.1% in water) was used in dose calculations. A dose rangefinder was performed using concentrations up to 5000 µg/plate of the test article in the presence and absence of 5.6% (v/v) S9-mix on S. typhimurium strain TA100 and E. coli strain WP2uvrA. The bacterial background lawn was not reduced and there was no biologically relevant decrease in the number of revertants which confirmed that this was an appropriate range of doses. The test article was then tested at 100 to 5000 µg/plate on S. typhimurium strains TA1535, TA1537 and TA98 in the presence and absence of 5% (v/v) S9-mix. An independent assay was performed at the same MTDID 6675 concentration range in the presence and absence of 10% (v/v) S9-mix on S. typhimurium strains TA1535, TA1537, TA98 and TA100 and E. coli strain WP2uvrA. The bacterial background lawn was not reduced and there was no biologically relevant decrease in the number of revertants in these assays. The test article did not precipitate on any plate in any assay. Negative control (Milli-Q water) and positive controls (strain specific) were included in each assay and the results indicated that the test conditions of each plate were adequate and the metabolic activation system functioned appropriately.The test article did not induce a significant dose-related increase in the number of revertant colonies in any of the tested strains both in the presence and absence of S9-metabolic activation. These results were confirmed in an independently repeated experiment. Based on the results of this study,the test article is not mutagenic in the Salmonella typhimurium reverse mutation assay and the Escherichia coli reverse mutation assay.