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Toxicological information

Genetic toxicity: in vitro

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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)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report Date:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
July 21, 1997
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch no. 1190567/006 provide by Raschig GmbH
- Expiration date of the lot/batch: 31.08.2001
- Purity test date: not available

RADIOLABELLING INFORMATION (if applicable): not available

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: stable
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: soluble (in water)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: none

TREATMENT OF TEST MATERIAL PRIOR TO TESTING: none

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 97
Species / strain / cell type:
S. typhimurium TA 98
Species / strain / cell type:
S. typhimurium TA 100
Species / strain / cell type:
S. typhimurium TA 102
Species / strain / cell type:
S. typhimurium TA 1535
Metabolic activation:
with and without
Metabolic activation system:
S9
Test concentrations with justification for top dose:
First experiment: 0.05.; 0.15; 0.5; 1.5; 5.0 mg/plate, where 5 mg/plate is the recommended maximum test concentration for soluble non-cytotoxic substances
Second experiment: 1.25; 2.5; 5.0 mg/plate, where 5 mg/plate is the recommended maximum test concentration for soluble non-cytotoxic substances
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
sodium azide
benzo(a)pyrene
other: 4-Nitro-1,2-phenylene diamine and 2-Amino-anthrcene
Details on test system and experimental conditions:
METHOD OF APPLICATION: 1st experiment: in agar (plate incorporation); 2nd experiment: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h (both experiments)
- Expression time (cells in growth medium): not available
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable

SELECTION AGENT (mutation assays): not applicable

NUMBER OF REPLICATIONS: 4 plates per strain and dose

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: not applicable

NUMBER OF CELLS EVALUATED: not applicable

DETERMINATION OF CYTOTOXICITY
- Method: other: titre ratio
- Any supplementary information relevant to cytotoxicity: none

OTHER EXAMINATIONS: none

- OTHER: none
Evaluation criteria:
reproducible increase of revertant colonies per plate (factor >=2)
Statistics:
none

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 97
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
Key result
Species / strain:
S. typhimurium TA 98
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
Key result
Species / strain:
S. typhimurium TA 100
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
Key result
Species / strain:
S. typhimurium TA 102
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
Key result
Species / strain:
S. typhimurium TA 1535
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:
TEST-SPECIFIC CONFOUNDING FACTORS: not available

RANGE-FINDING/SCREENING STUDIES: none

HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%): not available

ADDITIONAL INFORMATION ON CYTOTOXICITY: none

Any other information on results incl. tables

In a study according to OECD TG 471 under GLP concidtions, the substance was not mutagenic in five strains (TA97, TA98, TA100, TA102, TA1535) with and without metabolic activation.

Applicant's summary and conclusion

Executive summary:

The mutagenic potential of the substance was investigated in a bacterial reverse mutation assay according to OECD TG 471 under GLP. The substance was soluble and non-cytotoxic up to 5 mg/plate, i.e. the recommended maximum concentration. The substance was tested in five strains (TA97, TA98, TA100, TA102, TA1535) with and without metabolic activation (S9).

In a first experiment, five concentrations (0.05; 0.15; 0.5; 1.5; 5.0 mg/plate) were tested for 48 hours in the plate incorporation method. In the second experiment the concentrations 1.25, 2.5 and 5.0 mg/plate were tested (48 h) in the pre-incubation method. The number of colonies were evaluated by eye.

While the positive controls were clearly mutagenic, the substance did not increase the number of colonies. Therefore, the substance is considered to be not mutagenic.