Registration Dossier

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:
2002
Report Date:
2002

Materials and methods

Test guideline
Qualifier:
according 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
Test material form:
solid

Method

Target gene:
his D, his , his G and tryp E
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S-9 mix
Test concentrations with justification for top dose:
5 000 µg (recommended maximum test concentration)- 1 500 µg - 500 µg - 150 µg - 50 µg
Vehicle / solvent:
paraffin oil
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
9-aminoacridine
2-nitrofluorene
sodium azide
other: B-Propiolactone, 2-Anthramine, cis-platinium
Details on test system and experimental conditions:
For each of the strains, 100 µL of the bacterial suspension (1-5 x 109 bacteria/mL) and 100 µL of the test substance are mixed with 2.0 mL of overlay agar and poured over the surface of a minimal agar
plate (90 mm in diameter) (n = 3). The overlay agar is allowed to solidify before incubation.
Plates are incubated at 37° C over a 48 hour period. The number of revertant colonies per plate is counted.

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks:
b-propiolactone for test without metabolic activation. 2-anthramine for test with metabolic activation
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks:
9-aminoacridine for test without metabolic activation. 2-anthramine for test with metabolic
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks:
2-nitrofluorène for test without metabolic activation. 2-anthramine for test with metabolic
Key result
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks:
Sodium azide for test without metabolic activation. 2-anthramine for test with metabolic
Key result
Species / strain:
E. coli WP2 uvr A pKM 101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks:
Cis-platinum for test without metabolic activation. Dimethyl Benzanthracene for test with metabolic

Applicant's summary and conclusion

Conclusions:
There is no evidence of any increase in the number of revertant colonies in the presence of the test substance (5 000 µg - 1 500 µg - 500 µg - 150 µg - 50 µg) without and with metabolic activation.
Executive summary:

The genetox toxicity in vitro was performed according to the OECD 471 guideline.

The test substance does not induce any mutagenic change in Salmonella typhimarium TA1535, TA1537, TA98, TA100 and Escherichia coli WP2, with or without metabolic activation.