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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study out of Japan.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report Date:
2001

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 3-Cyanopyridine
- Molecular formula (if other than submission substance):
- Molecular weight (if other than submission substance): 104.12
- Smiles notation (if other than submission substance):
- InChl (if other than submission substance):
- Structural formula attached as image file (if other than submission substance): see Fig.
- Substance type: white solid
- Physical state: solid
- Analytical purity: 99.9%
- Impurities (identity and concentrations):
- Composition of test material, percentage of components:
- Isomers composition:
- Purity test date:
- Lot/batch No.: 709S4067
- Expiration date of the lot/batch:
- Radiochemical purity (if radiolabelling):
- Specific activity (if radiolabelling):
- Locations of the label (if radiolabelling):
- Expiration date of radiochemical substance (if radiolabelling):
- Stability under test conditions: stable under normal handling conditions and during period of test
- Storage condition of test material:
- Other: soluble in water

Method

Target gene:
histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Obtained from Professor B.N. Ames of the University of California on May 27, 1983. Each of the bacteria strains was kept in frozen storage in an ultra-low temperature cessel at -80 degrees C or lower.
Species / strain / cell type:
E. coli WP2 uvr A
Details on mammalian cell type (if applicable):
Obtained from Professor Matsushima of the Medical Research Institute of Tokyo University on October 10, 1985. Each of the bacteria strains was kept in frozen storage in an ultra-low temperature cessel at -80 degrees C or lower.
Metabolic activation:
with and without
Metabolic activation system:
S9 fraction from liver of induced rats.
Test concentrations with justification for top dose:
313, 625, 1250, 2500, 5000 micromole/plate (common ratio of 2). Maximum concentration of 50 mg/mL of 3-cyanopyridine in injection-use water, the solution was diluted with the same solvent at a common ration of 2 for use.
Vehicle / solvent:
Water
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
water
Positive controls:
yes
Positive control substance:
other: acrylamide
Remarks:
TA-98 and TA-100 without S9
Negative solvent / vehicle controls:
yes
Remarks:
water
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA-1535 without S9
Negative solvent / vehicle controls:
yes
Remarks:
water
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
WP2 uvr A without S9
Negative solvent / vehicle controls:
yes
Remarks:
water
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA-1537 without S9
Negative solvent / vehicle controls:
yes
Remarks:
water
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene
Remarks:
All strains with S9
Details on test system and experimental conditions:
The test was conducted in duplicate by the preincubation method. Into a test tube was poured 0.1 mL of test substance solution, 0.5 mL of 0.1 M sodium phosphate buffer solution (pH 7.4) and 0.1 mL of bacterial suspension were added, and the mixture was shaken for 20 min at 37 degrees C. When S9 was present, 0.5 mL of S9 mix was added instead of the 0.1 M sodium phosphate buffer solution. Following preincubation, 2 mL of top agar was admixed to the test tube and the contents tereof were layered on a minimum glucose agar plate medium. Once the layered top agar had solidified, the bacteria were cultured for 48 hours at 37 degrees C. The development of bacterial colonies was observed with a steromicroscope. After checking for the presence of antibackterial properties due to the test substance, the number of reverse mutation colonies on the plate was counted with an automatic colony counter. In the reliminaty tests, one plate was employed at each concentration. In the main test, three plates were employed at each concentration and the test was conducted twice to confirm reproducibility. Further, a negative control substance (solvent) and a positive control substance for each bacteria strain were employed in place of the test substance solution to provide control groups that were handled in the same manner as the test substance groups.
Evaluation criteria:
For each of the test bacteria strains, irrespective of the presence or absence of S9 mix, when the number of reverse mutation colonies (average value) accompanying an increase in test substance concentration increased to twice or more that of the negative control value, and that increase was found to be reproducible, the test substance was judged to be positive. All other cases were judged to be negative.
Statistics:
Statistical menthods were not employed to judge the test results.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
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:
In the results of two sets of main test conducted within the above-stated concentration range, no increase in the number of reverse mutation colonies of twice or more the level of the negative (solvent) control value was observed for any of the bacteria strains, irrespective of the presence or absence of S9 mix. Nor was any antibacterial property observed for any of the bacteria strains, irrespective of the presence or absence of S9 mix. Based on these results, it was concluded that the mutagenicity of 3-Cyanopyridine was negative.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

3-Cyanopyridine was tested in a standard Ames assay and no mutations were found with and without metabolic activation.