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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:
9 Sept 2010 - 21 October 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted under GLP conditions and OECD Guideline 471

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
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): MTDID 21721, TFEE-5
- Substance type: clear colorless liquid
- Physical state: liquid
- Lot/batch no.: TFEE5-276/12/09-1
- Expiration date of the lot/batch: 12 Dec 2012
- Stability under test conditions: stable
- Storage condition of test material: room temperature in the dark under nitrogen

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: Salmonella strains: histidine-dependent; E. coli: tryptophan-dependent
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
other: Salmonella strains: histidine-dependent; E. coli: tryptophan-dependent
Metabolic activation:
with and without
Metabolic activation system:
phenobarbital- and beta-naphthoflavone-induced rat liver S9
Test concentrations with justification for top dose:
First Experiment: 10, 33, 100, 333, 1000, 3330, and 5000 ug/plate (with and without S9)
Second Experiment: 33, 100, 333, 1000, and 3330 ug/plate (with and without S9)
Vehicle / solvent:
- Vehicle: ethanol
- Justification for choice of solvent/vehicle: The test substance was emulsified in ethanol at concentrations of 33.3 mg/ml and higher and was soluble in ethanol at concentrations of 10 mg/ml and lower; compatibility with the test system
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Without S9: TA1535, sodium azide; TA1537, 9-aminoacridine; TA98, 2-nitrofluorene; TA100, methylmethanesulfonate; WP2uvrA, 4-nitroquinoline N-oxide With S9: All strains, 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h
SELECTION AGENT (mutation assays): Salmonella strains: histidine; E. coli: tryptophan
NUMBER OF REPLICATIONS: 3 replicate plates in each experiment
DETERMINATION OF CYTOTOXICITY
- Method: reduction in the bacterial lawn, increase in the size of microcolonies, and decrease in the number of revertant colonies observed
Evaluation criteria:
A substance is considered positive (mutagenic) in the test if: a) the total number of revertants in tester strain TA100 is greater than 2-times the concurrent control, or the total number of revertants in tester strains TA 1535, TA 1537, TA 98, or WP2uvrA is greater than 3-times the concurrent control.
b) In case of a positive response, the positive response is reproducible in at least one independently repeated experiment.
Statistics:
Mean and SD of the number of revertant colonies per plate

Results and discussion

Test results
Species / strain:
other: S. typhimurium strains TA98, TA100, TA1535, and TA 1537 and E. coli strain WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: 3300 and 5000 ug/plate
COMPARISON WITH HISTORICAL CONTROL DATA: All control values were within the laboratory historical control ranges with the exception of the positive control value for tester strain TA98 in the absence of S9 in the first experiment (1264) which was slightly above the upper limit of the historical range (1260). This did not adversely affect the study integrity.
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 Criteria used for interpretation of results:OECD GHS

Based on the results of this study, TFEE-5 is not mutagenic in either the presence or absence of metabolic activation.
Executive summary:

The mutagenic potential of TFEE-5 (MTDID 21721, lot TFEE5-276/12/09-1) was evaluated in the Bacterial Reverse Mutation Assay with S. typhimurium strains TA98, TA100, TA1535 and TA1537 and E. coli strain WP2 uvrA in both the presence and absence of a metabolic activation system (S9-mix: phenobarbital- and beta-naphthoflavone-induced rat liver).

METHODS: This study was performed in compliance with OECD GLP (1997). The homogeneity, stability and concentration of the test substance were not analyzed. The study design was based on OECD 471 (1997), EC 440/2008 B13/14 L142 (2008) and Japan MHLW guidelines (2004). TFEE-5 was prepared in 100% ethanol for addition to the test system. TFEE-5 was tested at up to 5000 ug per plate in the first experiment and up to 3330 ug per plate in the second experiment. Both experiments were performed in the presence and absence of S9-mix. Strain-specific positive controls were included in all tests.

RESULTS: No increase in revertant colonies was observed in either the presence or absence of S9-mix for any strain. All criteria for a valid study were met as described in the protocol.

CONCLUSION: Under the conditions of this study, TFEE-5 was not mutagenic in the Bacterial Reverse Mutation Assay in either the presence or absence of metabolic activation.