Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In an Ames test the substance was tested in salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and E. colistrain WP2uvrAin presence and absence of metabolic activation. No increase in mutant frequency was reported in any of the strains tested both in presence and absence of metabolic activation. The substance is considered to be non-mutagenic under the conditions of this test.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 July 2017 to 29 September 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Qualifier:
according to
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Target gene:
histidine/tryptophan
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Details on mammalian cell type (if applicable):
CELLS USED
- Source of cells: University of California, Berkeley, on culture discs, on 04 August 1995, British Industrial Biological Research Association, on a nutrient agar plate, on 17 August 1987.
- Methods for maintenance in cell culture if applicable: stored at -196 °C in a Statebourne liquid nitrogen freezer, model SXR 34

Metabolic activation:
with and without
Metabolic activation system:
phenobarbital/beta-naphthaflavone induced rat liver S9 (Lot No. PB/BNF S9 20-08-2017 )
Test concentrations with justification for top dose:
Test 1 (plate incorporation): 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate
Test 2 (pre-incubation): 15, 50, 150, 500, 1500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: tetrahydrofurane
- Justification for choice of solvent/vehicle: substance is insoluble isterile distilled water, dimethyl sulphoxide, dimethyl formamide and acetonitrile at 50 mg/mL, acetone at 100 mg/mL and tetrahydrofuran at 200 mg/mL
A suspension in THF was considered as best doseable.
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
THF (in both tests <0.05 mL in final test solution)
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
N-ethyl-N-nitro-N-nitrosoguanidine
benzo(a)pyrene
other: 2-Aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: exp 1 plate incorporation; exp 2 preincubation

DURATION
- Preincubation period: 20 min at 37 ± 3 °C
- Exposure duration: 48 h at 37 ± 3 °C

NUMBER OF REPLICATIONS: 3/concentration

DETERMINATION OF CYTOTOXICITY
- Method: visible reduction in the growth of the bacterial background lawn
Evaluation criteria:
a substance is considered positive when:
1. A dose-related increase in mutant frequency over the dose range tested (De Serres and Shelby, 1979).
2. A reproducible increase at one or more concentrations.
3. Biological relevance against in-house historical control ranges.
4. Statistical analysis of data as determined by UKEMS (Mahon et al., 1989).
5. Fold increase greater than two times the concurrent solvent control for any tester strain (especially if accompanied by an out-of-historical range response (Cariello and Piegorsch, 1996)).
Statistics:
Dunnetts Regression Analysis
Key result
Species / strain:
other: TA1535, TA1537, TA98, TA100 and WP2uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
precipitate at 1500 ug/plate and above; in exp 2 reduced bacterial backgrownd lawn at 1500 ug/plate in TA 100 and TA1535 and at 5000 ug/plate in TA 100, TA1535 and TA98
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: the precipitate did not hinder scoring of the number of revertants
Conclusions:
Based on the findings in this test the substance is considered non-mutagenic in bacteria
Executive summary:

In an Ames test the substance was tested in salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and E. coli strain WP2uvrA in presence and absence of metabolic activation. No increase in mutant frequency was reported in any of the strains tested both in presence and absence of metabolic activation. The substance is considered to be non-mutagenic under the conditions of this test.

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

Additional information

Justification for classification or non-classification

Based on the information available, the substance does not need to be classified for mutagenicity according to Regulation (EC) No 1272/2008 (CLP).