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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Target gene:
his operon (for S. typhimurium strains)
trp operon (for E. coli strain)
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 5,6-benzoflavone
Test concentrations with justification for top dose:
without metabolic activation, all strains: 156, 313, 625, 1250, 2500, 5000 µg/plate
with metabolic activation, S. typhimurium strains: 156, 313, 625, 1250, 2500, 5000 µg/plate
with metabolic activation, E. coli strain: 313, 625, 1250, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle used: DMSO (0.1 mL per test)
- Justification for choice of vehicle: due to limited solubility of the test substance in water, DMSO was selected as vehicle.
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF2; 0.01 or 0.1 µg/plate, -S9, TA 100, TA 98, E. coli); sodium azide (SA; 0.5 µg/plate, -S9, TA 1535); 9-aminoacridine (9AA; 80 µg/plate, -S9, TA 1537); 2-Aminoanthracene (2AA; 0.5-10 µg/plate, all strains)
Details on test system and experimental conditions:
METHOD OF APPLICATION: Pre-incubation method

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3 replications each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition
Evaluation criteria:
Colonies were counted for 3 plates. Mean values which are at least twice as high as the negative vehicle control are considered as positive. Otherwise it is considered as negative.
Statistics:
Mean values and standard deviation 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:
cytotoxicity
Remarks:
at 2500 and 5000 µg/plate with and without S9 mix
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 2500 and 5000 µg/plate without S9 mix
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: YES; A "cytotoxicity" study was performed with 50, 150, 500, 1500 and 5000 µg/plate with and without S9 mix. Growth inhibition was observed at 5000 µg/plate without S9 mix in all tested strains (TA 98, TA 100, TA 1535, TA 1537 and E. coli WP2 uvrA) and at 5000 µg/plate with S9 mix in all S. typhimurium strains.

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 test substance did not induce gene mutations in S. typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and in E. coli WP2 uvrA strain at concentrations up to 5000 µg/plate with and without metabolic activation. Cytotoxicity was observed at and above 2500 µg/plate in all strains without S9 mix and in S. typhimurium strains with S9 mix.