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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:
From 9 MAY 1984 to 11 MAY 1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Basic data given: comparable to guidelines

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1984
Report date:
1984

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
o-anisidine
EC Number:
201-963-1
EC Name:
o-anisidine
Cas Number:
90-04-0
Molecular formula:
C7H9NO
IUPAC Name:
2-methoxyaniline
Details on test material:
- Name of test material (as cited in study report): Echtrot BB Base flüssig

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
other: auxotrophic for histidine
Species / strain / cell type:
S. typhimurium TA 1538
Additional strain / cell type characteristics:
other: auxotrophic for histidine
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
other: auxotrophic for tryptophan
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9 (Aroclor 1254 induced)
Test concentrations with justification for top dose:
0.004, 0.02, 0.1, 0.5, 2.5 and 10 µl/plate
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
Remarks:
, not required in this case
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene
Remarks:
with metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
Remarks:
, not required in this case
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene (all strains tested), 9-Aminoacridine (TA1537), Methylhydrazone derivative (TA98, TA1538, TA100), Streptocotocine (TA1535), ENNG (WP2 uvrA)
Remarks:
without metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 2 days

NUMBER OF REPLICATIONS:
- 4 plates per strain and dose level, including the negative controls
- only 2 plates per strain and dose level for positive controls
Evaluation criteria:
positive: number of revertants is more than double the spontaneous mutations

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
Vehicle controls validity:
not applicable
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not applicable
Untreated negative controls validity:
valid
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
Vehicle controls validity:
not applicable
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 with metabolic activation
negative without metabolic activation

The test item "Echtrot BB Base flüssig" did not excert mutagenic activity in the reverse bacterial mutation assay (plate incorporation, maximum dose level 10 µl/plate) with or without metabolic activation.
Executive summary:

This study was performed to investigate the potential of "Echtrot BB Base flüssig" to induce gene mutations in the plate incorporation test using the Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98, and TA 100, and the Escherichia coli strain WP2 uvrA. The assay was performed with and without liver microsomal activation. Each concentration, including the negative controls, was tested in quadruplicate. Positive controls were tested in duplicate. The test item was tested at the following concentrations: 0.004, 0.02, 0.1, 0.5, 2.5 and 10 µl /plate. No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), were observed. No substantial increase in revertant colony numbers of any of the six tester strains was observed following treatment with "Echtrot BB Base flüssig" at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance. Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies. In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.