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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
The test item was tested in two Ames Test. The test item is negative in Ames test.
Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1997-03-08 to 1999-03-1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline study
Reference:
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
May 1983/revised Draft, March 1996
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Version / remarks:
May 1983/revised Draft, March 1996
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
December 1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
testing lab.
Type of assay:
bacterial reverse mutation assay
Test material information:
Composition 1
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
Aroclor-induced rat liver S-9 mix
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Aroclor-induced rat liver S-9 mix
Test concentrations with justification for top dose:
20 ug - 5000 ug/plate (standard plate test)
4 ug - 2500 ug/plate (preincubation test)
Vehicle:
Due to the good solubility of the test substance in water, water was selected as the vehicle.
Negative controls:
yes
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: with metabolic activation: 2-aminoanthracene; without metabolic activation: N-methyl-N'-nitro-N-nitrosoguanidine, 4-nitro-o-phenylendiamine, 9-aminoacridine and N-ethyl-N'-nitro-N-nitrosoguanidine
Details on test system and conditions:
METHOD OF APPLICATION: Standard plate test (experiment I), preincubation (experiment II)

DURATION
- Preincubation period: 20 min (Experiment II)
- Exposure duration: 30 sec.
- Expression time: 48-72 hours

NUMBER OF REPLICATIONS: 3 test plates per dose or per control


Evaluation criteria:
The test chemical is considered positive in this assay if the following criteria are met :
• A dose-related and reproducible increase in the number of revertant colonies, i .e . about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.
A test substance is generally considered nonmutagenic in this test if :
• The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments
carried out independently of each other.
Statistics:
not required
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
A bacteriotoxic effect was observed under all test conditions.
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
A bacteriotoxic effect was observed under all test conditions.
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S-9 mix or after the addition of a metabolizing system. No test substance precipilation was found.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

In the key study, the test item was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several bacterial

strains, i .e . Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and Escherichia coli E . coli WP2 uvrA, in a reverse mutation assay. Standard plate test (SPT) and preincubation test (PIT) both with and without metabolic activation (Aroclor-induced rat liver S-9 mix). The dose range in the standard plate test was 20 µg - 5000 µg/plate. According to the results of the present study, the test item is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay.

In the supporting study, the test item was tested in Salmonella typhimurium TA1535 in an Ames test. The test item is negative without metabolic activation.


Justification for selection of genetic toxicity endpoint
Only one key study is available.

Justification for classification or non-classification

Based on the results, the substance was not classified according to Regulation (EC) No 1272/2008 and Directive 67/548/EEC (DSD).