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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: This study was selected as a key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labeling and/or risk assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
Qualifier:
according to
Guideline:
other: EC Commission Directive 2000/32/EC, Annex 4D-B13/14 No. L136
Deviations:
no
Remarks:
Conducted according to guideline in effect at time of study conduct
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Purity: not reported as such

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:
Aroclor 1254-induced rat liver S9
Test concentrations with justification for top dose:
33.3, 66.7, 100, 333, 667, 1000, 3333, and 5000 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Water
- Justification for choice of solvent/vehicle: Based on the solubility of the test substance and compatibility with the target cells.
Controls
Untreated negative controls:
yes
Remarks:
Sterile water
Negative solvent / vehicle controls:
yes
Remarks:
Sterile water
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (TA1535, TA1537, TA100, WP2 uvrA +S9), benzo[a]pyrene (TA98 +S9), 2-nitrofluorene (TA98 -S9), sodium azide (TA100 and TA1535 -S9), Acridine mutagen ICR-191 (TA1537 –S9), 4-nitroquinoline-N-oxide (WP2 urvA -S9)
Remarks:
All positive controls were diluted with dimethyl sulfoxide (DMSO) except sodium azide, which was diluted with water.
Details on test system and experimental conditions:
METHOD OF APPLICATION: The plate incorporation method was applied. Treatment without activation was conducted by adding 100 μL of an overnight culture containing approximately 10E9 bacteria per millilitre to top agar supplemented with L-histidine and D-biotin or L-tryptophan. The components were mixed and poured onto a plate containing minimal agar. Treatments with activation were conducted as those without activation except that S9 mix was added to the bacteria/top agar mixture before it was poured onto a minimal glucose agar plate. The plates were incubated at approximately 37±2°C for approximately 57 to 68 hours.

DURATION
- Exposure duration: 57 to 68 hours

NUMBER OF REPLICATIONS: 2 trials with 3 treatments per concentration
Evaluation criteria:
For the test substance to be evaluated positive, it must cause a dose-related increase in the mean revertants per plate unless observed at the top dose level only. Data sets for tester strains TA1535 and TA1537 were judged positive if the increase in mean revertants at the highest numerical dose response was ≥ 3.0-fold the mean concurrent negative control value (vehicle control). Data sets for tester strains TA98, TA100 and WP2 uvrA were judged positive if the increase in mean revertants at the highest numerical dose response was ≥ 2.0-fold the mean concurrent negative control value (vehicle control).
Statistics:
Trials were evaluated independently. For each selected tester strain, the mean number of revertants and the standard deviation at each concentration with and without S9 activation 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:
no cytotoxicity
Vehicle controls validity:
valid
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:
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 in the absence and presence of S9 activation

The study and the conclusions which are drawn from it fulfil the quality criteria (validity, reliability, repeatability).
Negative when tested in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and in Escherichia coli WP2 uvr A in the absence and presence of S9 activation.
Executive summary:

The test substance was evaluated for mutagenicity in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and in Escherichia coli WP2 uvr A with and without an exogenous metabolic activation system (S9). The maximum concentration tested was 5000 µg/plate. No evidence of mutagenic activity was detected in either of two independent trials. In this study, the test substance was negative.