Registration Dossier

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:
January 21st 2011 - February 4th 2011
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 2-(N-benzyl-N-methylamino( ethyl acetoacetate
- Physical state: Liquid
- Purity test date: 95.5%
- Lot/batch No.: 112/2010
- Expiration date of the lot/batch: December 2011
- Stability under test conditions: stable
- Storage condition of test material: at room temperature, protected from light

Method

Target gene:
TA 98: his D 3052
TA 100: his G 46
TA 1535: his G 46
TA 1537: his C 3076
TA 102: G 428
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
EXPERIMENT I: 0.0316, 0.100, 0.316, 1.0, 2.5 and 5.0 uL/plate
EXPERIMENT II: 0.0100, 0.0316, 0.100, 0.316, 1.0, 2.5 and 5.0 uL/plate
Controls
Untreated negative controls:
yes
Remarks:
Aqua dest.
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
other: strain specific positive controls
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 60 minutes at 37 °C
- Exposure duration: 48 hours

DETERMINATION OF CYTOTOXICITY
- Method: by a clearing or rather diminution of the background lawn or a reduction in the number of revertants down to a mutation factor <0.5 in relation to the solvent control.
Evaluation criteria:
The mean value of revertant counts is divided by the mean values of the solvent control. The biological relevance of the result is the criterion for the interpretation of results.
Statistics:
A statistical evaluation of the results is not regarded as necessary

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
EXP I: at conc. >= 2.5 uL/plate (without metabolic activation) and at a conc. of 5.0 uL/plate (with metabolic activation) ; EXP II: at conc. >= 1.0 uL/plate (without metabolic activation) and at a conc. >= 2.5 uL/plate (with metabolic activation)
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:
EXP I: at conc. >= 2.5 uL/plate (without metabolic activation) and at a conc. of 5.0 uL/plate (with metabolic activation) ; EXP II: at conc. >= 1.0 uL/plate (without metabolic activation) and at a conc. >= 2.5 uL/plate (with metabolic activation)
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

2 (N-Benzyl-N-methylamino) ethyl acetoacetate is considered to be non-mutagenic in this bacterial reverse mutation assay.