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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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study conducted according to OECD Guideline and in compliance with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
1997
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
2000
Deviations:
no
GLP compliance:
yes
Type of assay:
mammalian cell gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

Method

Target gene:
thymidine kinase
Species / strain
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9-mix, Phenobarbital/ß-naphthoflavone induced
Test concentrations with justification for top dose:
Cytotoxicity test concentrations:
1.22, 4.88, 19.53, 78.13, 312.5, 1250 and 5000 µg/mL (3-hour treatment period with and without S9)
0.3125, 0.625, 1.25, 2.5, 5, 10 and 20 µg/mL (24-hour treatment period without S9)

Genotoxicity test concentrations:
1st experiment: 0, 0.625, 1.25, 2.5, 5, 10 and 20 µg/mL (3-hour treatment period, with and without S9)
2nd experiment: 0, 0.313, 0.625, 0.938, 1.25, 2.5 and 5 µg/mL (24-hour treatment period, without S9),
0, 2.5, 5, 10, 15, 20 and 30 µg/mL (3-hour treatment period, with S9)
Vehicle / solvent:
- Vehicle/solvent used: R0 medium
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: with S9-mix: Cyclophosphamide, without S9-mix: Ethylmethanesulphonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 24 hours and 3 hours

NUMBER OF REPLICATIONS: 2

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
Evaluation criteria:
Treated cultures were examined for a significant increase in mutant frequency.
Statistics:
UKEMS statistical package

Results and discussion

Test results
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
treatment period / test concentration / decrease in relative suspension growth: without S9-mix: 3 h / 20 µg/mL / 98 %; with S9-mix: 3 h / 20 µg/mL / 70 and 73 %; without S9-mix: 24 h / 5 µg/mL / 91 %
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxicity was observed at 20 µg/mL with and without S9-mix (3 hour treatment period) and at 5 µg/mL without S9 (24-hour treatment period)
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion