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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:
2007
Reliability:
1 (reliable without restriction)
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
GLP compliance:
yes
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Target gene:
Thymidine Kinase Locus (TK+/-) in Mouse Lymphoma L5178Y Cells
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9
Test concentrations with justification for top dose:
Pre-Test: 2.3, 4.7, 9.4, 18.8, 37.5, 75, 150, 300µg/ml with and without S9 mix
Experiment I: 18.8, 37.5, 75, 150, 300, 600 µg/ml with and without S9 mix;
Experiment II: 18.8, 37.5, 75, 150, 300, 600 µg/ml with and without S9 mix;
Vehicle / solvent:
DMSO
Controlsopen allclose all
Positive controls:
yes
Positive control substance:
cyclophosphamide
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Untreated negative controls:
yes
Negative solvent / vehicle controls:
other: medium
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium;

DURATION
- Exposure duration: 4, and 24 h
- Expression time (cells in growth medium): 48 to 72 h;
- Selection time (if incubation with a selection agent): approximately 11 d

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: approximately 4000

DETERMINATION OF CYTOTOXICITY
Relative Suspension Growth

OTHER EXAMINATIONS:
- Determination of mutant colony numbers
-small mutant colonies
-large mutant colonies

Evaluation criteria:
standard
Statistics:
A linear regression (least squares) was performed to assess a possible dose dependent increase of mutant frequencies using SYSTAT 11 software. The number of mutant colonies obtained for the groups treated with the test item was compared to the solvent control groups.

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
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
Additional information on results:
Precipitation at max. dose (600 µg/ml) visible even by naked eye;
COMPARISON WITH HISTORICAL CONTROL DATA: y
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

the test item did not induce mutations in the mouse lymphoma thymidine kinase locus assay using the cell line L5178Y in the absence and in the presence of metabolic activation; No substantial and reproducible dose dependent increase in mutant colony numbers was observed. No relevant shift of the ratio of small versus large colonies occurred.
The substance is not subject of classification and labelling