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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:
1996-01-01
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
Mutagenicity Test Data of Existing Chemical Substances
Author:
anonymous
Year:
1996
Bibliographic source:
Japan Chemical Industry Ecology Toxicology & Information Center, Japan (JETOC)
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
equivalent or similar to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

Method

Target gene:
The Salmonella typhimurium histidine (his) reversion system measures his- -> his+ reversions.
Species / strainopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Species / strain:
S. typhimurium TA 102
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Species / strain:
S. typhimurium TA 1538
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Species / strain:
other: TA104
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Species / strain:
E. coli WP2 uvr A
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Species / strain:
E. coli WP2 uvr A pKM 101
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction
Test concentrations with justification for top dose:
10, 20, 50, 100, 200, 500, 1000, 2000 µg/plate
Vehicle:
DMSO
Controlsopen allclose all
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -2-(2-furyl)-3-(5-nitro-2furyl)acrylamide
Remarks:
without activation: TA100, WP2uvrA and WP2uvrA/pKM101
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
without activation: TA1585
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without activation: TA1537
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
without activation: TA1538
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
without activation: TA1538
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: bleomycin
Remarks:
without activation: TA102
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: pyruvic aldehyde
Remarks:
without activation: TA104
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-amino anthracene
Remarks:
with activation: all cited strains (S. typhimurium TA 1535, TA 1537, TA1538, TA 98, TA 100, TA 102, TA 104, WP2uvrA and WP2uvrA/pKM101)
Evaluation criteria:
Two-fold rule criteria was used for data evaluation (Ames et al., 1975). The chemicals are considered to be mutagenic when a dose- related increase in revertant colonycount is observed and the number of the revertant colonies per plate with the test substance is more than twice that of the negative control (solvent control) and when the reproducibility of the test result is observed.
Mutagenic potency was calculated by the following equation and maximum value of mutagenic potency was expressed as the specific activity on the data sheet:
mutagenic potency (induced revertants / mg test substance) = (number of induced revertants on the dose X - number of revertant on the solvent control) / mg test chemical on the dose X.
Statistics:
Determination of Mean and Standard deviation

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity:
yes
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
other: TA 104
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Key result
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not applicable
Positive controls valid:
yes
Additional information on results:
Mutagenicity (frame-shift) was observed for S. typhimurium TA 98 and TA 1538 at concentrations of 10 to 1000 µg/plate. An evaluation of 2000 µg/plate was not possible due to growth inhibition.
The specific mutagenicity in case of TA 98 was 5900 revertants /mg, in case of TA 1538 13900 revertants /mg.

Applicant's summary and conclusion

Conclusions:
The reported data of this mutagenicity assay according to AMES shows, that after metabolic activation, the test item induced gene mutations by frameshift in the genome of 2 of 9 strains (S. typhimurium TA 98 and TA 1538) used. Therefore, TODI is considered mutagenic in this bacterial reverse mutation assay.
Executive summary:

A study was performed to assess the mutagenicity potency of TODI. The method was similar to OECD Guideline 471 (Bacterial Reverse Mutation Assay and EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria) of Commission Directive 92/69/EEC (which constitues Annex V of Council Directive 67/548/EEC). The test strains S. typhimurium TA 1535, TA 1537, TA 1538, TA 98, TA 100, TA 102, TA 104 and E. coli WP2 uvr A as well as E. coli WP2 uvr A pKM 101 were examined at 10, 20, 50, 100, 200, 500, 1000, 2000 µg TODI/plate. Mutagenicity (frame-shift) was observed for S. typhimurium TA 98 and TA 1538 at concentrations of 10 to 1000 µg/plate. An evaluation of 2000 µg/plate was not possible due to growth inhibition. The reported data of this mutagenicity assay according to AMES shows, that after metabolic activation, the test item induced gene mutations by frameshift in the genome of 2 of 9 (S. typhimurium TA 98 and TA 1538) strains used. Therefore, TODI is considered mutagenic in this bacterial reverse mutation assay.