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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:
January 2013 (study plan) through April 2013 (final report)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Principles of method if other than guideline:
PURPOSE AND PRINCIPLE OF THE STUDY
Cited from OECD guideline 471: “The bacterial reverse mutation test uses amino-acid requiring
strains of Salmonella typhimurium and Escherichia coli to detect point mutations,
which involve substitution, addition or deletion of one or a few DNA base pairs. The principle
of this bacterial reverse mutation test is that it detects mutations which revert mutations
present in the test strains and restore the functional capability of the bacteria to synthesize
an essential amino acid.”
The bacterial reverse mutation test is commonly employed as an initial screen for genotoxic
activity and, in particular, for point mutation-inducing activity.
Principle of the test method: Suspensions of bacterial cells are exposed to the test substance
in the presence and in the absence of an exogenous metabolic activation system.
In the plate incorporation method, these suspensions are mixed with an overlay agar and
plated immediately onto minimal medium. In the preincubation method, the treatment mixture
is incubated and then mixed with an overlay agar before plating onto minimal medium.
For both techniques, after two or three days of incubation, revertant colonies are counted
and compared to the number of spontaneous revertant colonies on solvent control plates.
This study was performed in order to evaluate the mutagenic potential of 4-
(TRIFLUOROMETHOXY) ANILINE in the Bacterial Reverse Mutation Test using five
strains of Salmonella typhimurium. The test was chosen on behalf of the sponsor.
Sponsor’s intent: Registration in accordance with: REACH
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
4-(trifluoromethoxy)aniline
EC Number:
207-317-5
EC Name:
4-(trifluoromethoxy)aniline
Cas Number:
461-82-5
Molecular formula:
C7H6F3NO
IUPAC Name:
4-(trifluoromethoxy)aniline
Test material form:
other: solution
Details on test material:
4-(TRIFLUOROMETHOXY) ANILINE, Batch no. KLP006B041209014

Method

Target gene:
see study report
Test concentrations with justification for top dose:
see study report
Vehicle / solvent:
see study report

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Findings and Results:
Two valid experiments were performed.
First Experiment:
Six concentrations of the test item, dissolved in dimethyl sulfoxide (DMSO) (ranging from
5003 to 15 μg/plate) were used. Five genetically manipulated strains of Salmonella typhimurium
(TA 97a, TA 98, TA 100, TA 102 and TA 1535) were exposed to the test item both
in the presence and in the absence of a metabolic activation system (S9) for 48 hours, using
the plate incorporation method.
None of the concentrations caused a significant increase in the number of revertant colonies
in the tested strains. The test item didn’t show any mutagenic effects in the first experiment.
Signs of toxicity towards all tested strains were observed in the highest concentration
(5003 μg/plate).
The sterility control and the determination of the titre didn’t show any inconsistencies. The
determined values for the spontaneous revertants of the negative controls were in the
normal range. All positive controls showed mutagenic effects with and without metabolic
activation.
Second Experiment:
To verify the results of the first experiment, a second experiment was performed, using six
concentrations of the test item (ranging from 1502 to 47 μg/plate) and a modification in
study performance (pre-incubation method).
The test item didn’t show mutagenic effects in the second experiment, either.
Signs of toxicity towards all tested strains could be observed in the highest concentration
(1502 μg/plate) again.
The sterility control and the determination of the titre didn’t show any inconsistencies. The
determined values for the spontaneous revertants of the negative controls were in the
normal range. All positive controls showed mutagenic effects with and without metabolic
activation.
Under the conditions of the test, the test item didn’t show mutagenic effects towards Salmonella
typhimurium, strains TA 97a, TA 98, TA 100, TA 102 and TA 1535.
Therefore, no concentration-effect relationship could be determined.
The test item 4-(TRIFLUOROMETHOXY) ANILINE is considered as
“not mutagenic under the conditions of the test”.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
The test item 4-(TRIFLUOROMETHOXY) ANILINE is considered as
“not mutagenic under the conditions of the test”.
Executive summary:

Findings and Results: Two valid experiments were performed. First Experiment: Six concentrations of the test item, dissolved in dimethyl sulfoxide (DMSO) (ranging from 5003 to 15 μg/plate) were used. Five genetically manipulated strains of Salmonella typhimurium (TA 97a, TA 98, TA 100, TA 102 and TA 1535) were exposed to the test item both in the presence and in the absence of a metabolic activation system (S9) for 48 hours, using the plate incorporation method. None of the concentrations caused a significant increase in the number of revertant colonies in the tested strains. The test item didn’t show any mutagenic effects in the first experiment. Signs of toxicity towards all tested strains were observed in the highest concentration (5003 μg/plate). The sterility control and the determination of the titre didn’t show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation. Second Experiment: To verify the results of the first experiment, a second experiment was performed, using six concentrations of the test item (ranging from 1502 to 47 μg/plate) and a modification in study performance (pre-incubation method). The test item didn’t show mutagenic effects in the second experiment, either. Signs of toxicity towards all tested strains could be observed in the highest concentration (1502 μg/plate) again. The sterility control and the determination of the titre didn’t show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range. All positive controls showed mutagenic effects with and without metabolic activation. Under the conditions of the test, the test item didn’t show mutagenic effects towards Salmonella typhimurium, strains TA 97a, TA 98, TA 100, TA 102 and TA 1535. Therefore, no concentration-effect relationship could be determined. The test item 4-(TRIFLUOROMETHOXY) ANILINE is considered as “not mutagenic under the conditions of the test”.