Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Specific details on test material used for the study:
Supplier: Changzou Foreign Trade Corp.Batch No.: 0041A25 (20141211)Physical state: solid substance, crystalline powderColour: whiteStorage in a cool, dry place, protected from light

Method

Target gene:
Salmonella typhimurium TA98, TA100, TA1535 and TA1537: GCEscherichia coli WP2 uvrA: AT
Species / strainopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9
Test concentrations with justification for top dose:
Based on the results of the preliminary Range Finding Test the following concentrations of the test item were prepared and used in the Initial and Confirmatory Mutation Tests: 5000, 1600, 500, 160, 50 and 16 μg/plate.
Vehicle:
Dimethyl sulfoxide (DMSO)Ultrapure water (UPW)
Controlsopen allclose all
Positive controls:
yes
Positive control substance:
other: 4-Nitro-1,2-phenylenediamine, NPD
Remarks:
4 μg/plate, Salmonella typhimurium TA98, without metabolic activation
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
2 μg/plate, Salmonella typhimurium TA100 and TA1535, without metabolic activation
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
50 μg/plate, Salmonella typhimurium TA1537, without metabolic activation
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
2 μL/plate, Escherichia coli WP2 uvrA, without metabolic activation
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
2 μg/plate for S. typhimurium strains; 50 μg/plate for E. coli WP2 uvrA, Salmonella typhimurium TA98; TA100; TA1535; TA1537 and Escherichia coli WP2 uvrA, with metabolic activation [10 % (v/v) rat liver]
Negative controls:
yes
Remarks:
vehicle Control
Remarks:
DMSO, UPW
Details on test system and conditions:
The study included a Preliminary Solubility Test, a Preliminary Range Finding Test (Informatory Toxicity Test) an Initial Mutation Test (Plate Incorporation Test) and a Confirmatory Mutation Test (Pre-Incubation Test). In the preliminary Range Finding Test as well as in the Initial Mutation Test the plate incorporation method was used. Based on the results of the preliminary tests, a concentrated stock solution was prepared from the test item with Dimethyl sulfoxide (DMSO), which was diluted by serial dilutions to obtain the six dosing solutions investigated in the main tests. The maximum test concentration was in all examined strains 5000 μg test item/plate (±S9 Mix). The AMIDINOTHIOUREA (CAS No. 2114-02-5) concentrations investigated in the Initial and Confirmatory Mutation Tests: 5000, 1600, 500, 160, 50 and 16 μg/plate. Test solutions were freshly prepared at the beginning of the experiment.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to limit concentrations
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes

Applicant's summary and conclusion

Conclusions:
In conclusion, the test item AMIDINOTHIOUREA (CAS No. 2114-02-5) has no mutagenic activity on the applied bacterium tester strains under the test conditions used in this study.