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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Conducted at GLP-accredited facility to current guidelines

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
10% rat liver S9
Test concentrations with justification for top dose:
0, 50, 150, 500, 1500, 5000 µg/plate
Vehicle / solvent:
DMSO - test material miscible
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
without activation
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without activation
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
without activation
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
with activation
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
Remarks:
with activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: none
- Exposure duration: 48 hours


NUMBER OF REPLICATIONS: 2 experiments, with each strain/concentration performed in triplicate with/without activation


DETERMINATION OF CYTOTOXICITY
- Method: background lawn

ACCEPTANCE CRITERIA
All tester strains exhibit a characteristic number of spontaneous revertants in the vehicle and controls
Appropriate characteristics for each tester strain confirmed
All tester strain cultures in the range 1-9.9x10^9 bacteria per ml
Each mean positive control should be at least two-fold higher than the vehicle control
A minimum of 4 non-toxic test material concentrations
No evidence of excessive contamination
Evaluation criteria:
Dose-related increase in revertants over the concentration range or a reproducible increase at one or more concentrations in at least one strain with or without activation
Biological relevance considered first, with statistical methods used as an aid to evaluation but not the only factor in determining a positive response
Statistics:
As recommended by UKEMS

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:
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
Species / strain:
E. coli WP2 uvr A
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
The test material was not mutagenic to any tester strain, either in the presence or absence of metabolic activation, up to the recommended limit concentration.