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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
Not mutagenic in the Salmonella typhimurium reverse mutation assay (OECD guideline 471; GLP)
Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1995-01-24 to 1995-03-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
revised draft document of May 1994
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
31 July 1992
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
his
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
preliminary toxicity test: 0, 10, 100, 500, 1000, 2500 and 5000 µg/plate
mutagenicity tests: 0, 312.5, 625, 1250, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
mitomycin C
Remarks:
without metabolic activation
Positive controls:
yes
Positive control substance:
other: 2-anthramine, Danthron
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); second test with metabolic activation: preincubation

DURATION
- Preincubation period: 60 min (second test with metabolic activation)
- Exposure duration: 48-72 h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: background bacterial lawn
Evaluation criteria:
The following criteria were used as an aid for determining a positive response:
- a reproducible and significant dose relationship
and/or
- a reproducible and significant increase (i.e. a doubling in the number of revertants for at least one of the tested strains when compared to that of the controls) for at least one of the doses

A test substance is considered as non-mutagenic in this test system if the above two criteria are not fully met.
Biological and statistical significances were considered during the evaluation.
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no

COMPARISON WITH HISTORICAL CONTROL DATA:
controls were within historical range

ADDITIONAL INFORMATION ON CYTOTOXICITY:
slight to moderate cytotoxicity was observed in all strains at 5000 µg/plate without metabolic activation; no cytotoxic effects were seen with metabolic activation up to the limit dose
Conclusions:
TFMEA did not induce mutant colonies over background. TFMEA was tested up to limit concentrations of 5000 µg/plate.
Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471, revised draft document of May 1994, S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains were exposed to TFMEA (99.94% a.i.) in DMSO at concentrations of 0, 312.5, 625, 1250, 2500 and 5000 µg/plate in the presence and absence of mammalian metabolic activation. The test was performed as plate incorporation assay; the second experiment with metabolic activation was performed as pre-incubation test with 60 minutes pre-incubation. TFMEA was tested up to limit concentrations (5000 µg/plate). Only slight cytotoxic effects were observed in the highest concentration without metabolic activation.

The positive controls induced the appropriate responses in the corresponding strains. 

There was no evidence of induced mutant colonies over background. 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

One reliable (RL=1), relevant and adequate study to assess the genotoxic potential of TFMEA is available:

In a reverse gene mutation assay in bacteria according to OECD guideline 471, revised draft document of May 1994, S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 strains were exposed to TFMEA (99.94% a.i.) in DMSO at concentrations of 0, 312.5, 625, 1250, 2500 and 5000 µg/plate in the presence and absence of mammalian metabolic activation. The test was performed as plate incorporation assay; the second experiment with metabolic activation was performed as pre-incubation test with 60 minutes pre-incubation. TFMEA was tested up to limit concentrations (5000 µg/plate). Only slight cytotoxic effects were observed in the highest concentration without metabolic activation.

The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background. 

 

Thus, based on the available information, TFMEA is not genotoxic. There are no data gaps for the endpoint genotoxicity. No human information is available for this endpoint. However, there is no reason to believe that these results would not be applicable to humans.


Justification for selection of genetic toxicity endpoint
OECD guideline study; GLP

Justification for classification or non-classification

Based on the available data TFMEA does not need to be classified for mutagenicity according to the criteria given in regulation (EC) 1272/2008 or the former European directive on classification and labelling 67/548/EEC. Thus, no labelling is required.