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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Test material form:
liquid

Method

Species / strain
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 liver microsomal fraction and S9 Mix
Test concentrations with justification for top dose:
The test item concentrations to be applied in the main experiments were chosen according to the results of the pre-experiment. 5000 μg/plate was selected as the maximum concentration. The concentration range covered two logarithmic decades. Two independent experiments were performed with the following concentrations:
Experiment I:
10.0, 31.6, 100, 316, 1000 and 2500 μg/plate
Experiment II:
5.0, 15.8, 50.0, 158, 500, 1580 and 5000 μg/plate
Vehicle / solvent:
The test item was dissolved in Ethanol and diluted prior to treatment. The solvent was compatible with the survival of the bacteria and the S9 activity.
Controls
Untreated negative controls:
yes
Remarks:
A. dest., Eurofins Munich, Lot No. 170220
Negative solvent / vehicle controls:
yes
Remarks:
Ethanol, AppliChem Lot No. 5Y103701
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-nitro-o-phenylene-diamine; 2-aminoanthracene

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
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:
S. typhimurium TA 1537
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:
S. typhimurium TA 102
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:
S. typhimurium 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:
S. typhimurium TA 98
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

Applicant's summary and conclusion

Conclusions:
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, TEA Trioleate did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used.
Therefore, TEA Trioleate is considered to be non-mutagenic in this bacterial reverse mutation assay.
Executive summary:

The test item TEA Trioleate was investigated for its potential to induce gene mutations according to the plate incorporation test (experiment I and II) using Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 102.

In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate. The following concentrations of the test item were prepared and used in the experiments:

Experiment I:

10.0, 31.6, 100, 316, 1000 and 2500 μg/plate

Experiment II:

5.0, 15.8, 50.0, 158, 500, 1580 and 5000 μg/plate

Precipitation of the test item was observed in all tester strains used in experiment I and II (with and without metabolic activation). In experiment I precipitation of the test item was found at concentrations of 1000 μg/plate and higher (with and without metabolic activation). In experiment II precipitation of the test item was found at a concentration of 5000 μg/plate (with and without metabolic activation).

Toxic effects of the test item were noted in some tester strains evaluated in experiment I and II.

In experiment I toxic effects of the test item were observed in tester strain TA 100 at concentrations of 1000 μg/plate and higher (without metabolic activation). In tester strain TA 1535 toxic effects of the test item were noted at a concentration of 2500 μg/plate (without metabolic activation). In tester strain TA 1537 toxic effects of the test item were observed at a concentration of 2500 μg/plate (with metabolic activation).

In experiment II toxic effects of the test item were noted in tester strain TA 1537 at concentrations of 1580 μg/plate and higher (without metabolic activation).

No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with TEA Trioleate at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II.

All criteria of validity were met.