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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Oct. 15, 2008 ~ Dec. 31, 2008
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
yes (incl. certificate)
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Everzol Supra Navy Blue FBN
- Molecular formula (if other than submission substance): C37H23ClN10O22S7. 6Na
- Molecular weight (if other than submission substance): 1356.5
- Smiles notation (if other than submission substance): not applicable
- InChl (if other than submission substance): not applicable
- Structural formula attached as image file (if other than submission substance): see section 1.2
- Substance type: mono constituent substance
- Physical state: powder
- Analytical purity: 72.1%
- Impurities (identity and concentrations): see section 1.2
- Composition of test material, percentage of components: see section 1.2
- Isomers composition: see section 1.2
- Lot/batch No.: SR328515
- Stability under test conditions: stable
- Storage condition of test material:2 ~ 8 ℃

Method

Target gene:
thymidine kinase
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
- Type and identity of media: RPMI1640
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254-induced post-mitochondrial fraction (S9)
Test concentrations with justification for top dose:
5, 4.25, 3.5, 3 mg/mL for non-activation system
5, 4.25, 3.5, 3, 2.5, 1.25, 0.625 mg/mL for non-activation system
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: Everzol Supra Navy Blue FBN is soluble in water up to 100g/L

Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
2-acetylaminofluorene
Remarks:
Migrated to IUCLID6: ethylmethanesulphonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium


DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 12 days


SELECTION AGENT (mutation assays): Trifluorothymidine (TFT)



DETERMINATION OF CYTOTOXICITY
- Method: viability, relative suspension growth



Evaluation criteria:
Mutant Frequency

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
positive
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:
Interpretation of results (migrated information):
positive

According to Clive (1987), "twofold" rule of M.F. is a criterion for judging positive response. In the non-activation system, the M.F. of 5 and 4.25 mg/mL of Everzol Supra Navy Blue FBN were approximately twofold than negative control. In the S9 activation system, the M.F. of 5, 4.25, 3.5 and 3 mg/mL of Everzol Supra Navy Blue FBN were approximately twofold than negative control.