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Genetic toxicity in vitro

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:
03 May 2012 to 14 May 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted under GLP conditions
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
In 2nd exp in tester strain TA100,only 2 plates of test substance conc 3330 μg/plate in presence of S9-mix were tested.All other dose levels were plated in triplicate & testing of extra plate would have given no additional info: deviation had no influence
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
yes
Remarks:
In 2nd exp in tester strain TA100,only 2 plates of test substance conc 3330 μg/plate in presence of S9-mix were tested.All other dose levels were plated in triplicate & testing of extra plate would have given no additional info: deviation had no influence
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9-fraction
Test concentrations with justification for top dose:
100, 333, 1000, 3330, and 5000 ug test article per plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: dimethyl sulfoxide (DMSO) (SeccoSolv, Merck, Darmstadt, Germany)
Untreated negative controls:
yes
Remarks:
Vehicle used as a negative control.
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: Without metabolic act TA1535: sodium azide (SA) TA1537: 2-nitrofluorene (NF) TA98: 2-nitrofluoroene (NF) TA100: methylmethanesulfonate (MMS) WP2uvrA: 4-nitroquinoline N-oxide (4-NQO) With metabolic activation All strains: 2-aminoanthracene (2AA)
Details on test system and experimental conditions:
METHOD OF APPLICATION:Test substance was included in the top agar.
DURATION
- Preincubation period:
- Exposure duration: 48 hours +/- 4 hours.
STAIN (for cytogenetic assays):
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
Evaluation criteria:
The negative control data (number of spontaneous revertants per plate) should be within the
laboratory historical range for each tester strain. Also, the positive control chemicals should produce responses in all tester strains, which are within the laboratory historical range documented for each positive control substance. Furthermore, the mean plate count should be at least three times the concurrent vehicle control group mean.
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and 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
Species / strain:
E. coli WP2 uvr A
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative EU

No increase in revertant colonies was observed in either the presence or absence or S9 mix for any strain. All criteria for a valid study were met as described in the protocol. Under the conditions of the study, the test article was not mutagenic in the Bacterial Reverse Mutation Assay in either the presence or absence of metabolic activation.
Executive summary:

The mutagenic potential of the test article (clear colorless liquid, purity 99.99%) was evaluated in the Bacterial Reverse Mutation Assay with S. typhimurium strains TA98, TA100, TA1535, and TA1537 and E. coli strain WP2 uvrA in the presence and absence of a metabolic activation system (S9-mix rat liver induced by Phenobarbital and B-napthoflavone). The study was performed in compliance with OECD GLP (1997). The study design was based on OECD No. 471 (1997) and EC 440/2008 B.12/14 L142 (2008). The test article was dissolved in dimethyl sulfoxide. A dose range-finding test was performed at concentrations up to 5000ug/plate in the absence and presence of S9 mix in strains TA100 and WP2uvrA. A second assay was performed in all strains at doses of 100, 333, 1000, 3330, and 5000 ug/plate in the presence or absence of S9 mix. Strain specific positive controls and vehicle controls were tested in parallel. All treatments were performed in triplicate. No increase in revertant colonies was observed in either the presence or absence or S9 mix for any strain. All criteria for a valid study were met as described in the protocol. Under the conditions of the study, the test article was not mutagenic in the Bacterial Reverse Mutation Assay in either the presence or absence of metabolic activation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Justification for classification or non-classification

Criteria for classifying CAS# 756-12-7 as mutagenic are not met.