Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Bacterial Reverse Mutation Test

The test substance is not mutagenic in the bacterial reverse mutation test, either in the absence or presence of a metabolic activation system.

Chromosome Abberation Test

The test substance did not induce either structural or numerical chromosome aberrations in the absence or presence of a metabolic activation system.

HPRT Test

Under the experimental conditions of this study, the test substance is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation.

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

Genetic toxicity in vivo

Description of key information

Micronucleus Assay

It was concluded that the test item did not induce micronuclei in the bone marrow cells of NMRI mice.

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

Additional information

Genetic Toxicity in vitro

 

Key study: Bacterial reverse mutation assay 2015/1099097

The test substance was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli (WP2uvrA). The test was performed in two experiments in the presence and absence of S9-mix (rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone) using the standard plate and the pre-incubation methods. The test substance was dissolved in dimethyl sulfoxide. The test substance was tested in both experiments at concentrations of 33, 100, 333, 1000, 2600 and 5200 μg/plate.

All bacterial strains showed negative responses over the entire dose range, i.e. no significant dose-related increase in the number of revertants in two independently repeated experiments. The positive controls induced the appropriate responses in the corresponding strains. The test substance is not mutagenic in the bacterial strains tested, either in the absence or presence of a metabolic activation system.

 

Supporting studies: Bacterial reverse mutation assay

All studies are GLP and guideline compliant.

In the supporting studies the test substance was also determined to be not mutagenic in the bacterial strains tested, either in the absence or presence of a metabolic activation system.

 

 

Key study: Chromosome aberration test 2009/8000063

The test substance was tested in the in vitro cytogenetic test using cultured Chinese hamster lung (CHL) cells. The test was performed in two independent experiments without and with a metabolic activation system (rat liver S9-mix induced by a combination of phenobarbital and ß-naphthoflavone). The test substance was dissolved in dimethyl sulfoxide. In the short-term treatment, the test substance was treated at a concentration of 78.1, 156, 313 or 625 μg/ml for 6 hours with or without metabolic activation. Eighteen hours after the termination of the treatment, chromosome preparations were made. In the continuous treatment, the test substance was treated without metabolic activation at a concentration of 30.9, 46.3, 69.4, 104 or 156 μg/ml for 24 hours; severe toxicity at the two highest doses prevented analysis. As a result of the metaphase analysis in the cytogenetics tests both with the short-term (with and without metabolic activation) and the continuous treatment (without metabolic activation), there were no significant increases in the frequencies of the metaphases with structural chromosome aberrations and polyploid metaphases at any concentrations of the test substance when compared with concurrent solvent control.

Positive control chemicals, mitomycin C and benzo(a)pyrene, both produced a statistically significant increase in the incidence of cells with chromosome aberrations. The test substance did not induce either structural or numerical chromosome aberrations in the absence or presence of a metabolic activation system.

 

 

Key study: HPRT 2015/111062

The substance was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro. Two independent experiments were carried out, both with and without the addition of liver S9 mix from phenobarbital- and ß-naphthoflavone induced rats (exogenous metabolic activation).

According to an initial range-finding cytotoxicity test for the determination of the experimental doses the following concentrations were tested.

1st Experiment

without S9 mix : 0; 4.7; 9.4; 18.8; 37.5; 75.0; 150.0; 300.0 μg/mL

with S9 mix : 0; 2.3; 4.7; 9.4; 18.8; 37.5; 75.0; 150.0; 300.0 μg/mL

2nd Experiment

without S9 mix : 0; 3.9; 7.8; 15.6; 31.3; 62.5; 125.0; 250.0 μg/mL

with S9 mix : 0; 3.9; 7.8; 15.6; 31.3; 62.5; 125.0; 250.0 μg/mL

The vehicle controls gave mutant frequencies within the range expected for the CHO cell line. Both positive control substances, EMS and DMBA, led to the expected increase in the frequencies of forward mutations.

In both experiments in the absence and in the presence of metabolic activation no cytotoxicity was observed up to the highest tested concentration evaluated for gene mutations. Based on the results of the present study, the test substance did not cause any relevant increase in the mutant frequencies either without S9 mix or after the addition of a metabolizing system in two experiments performed independently of each other.

Thus, under the experimental conditions of this study, the test substance is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation.

Genetic Toxicity in vivo

Key study: Micronucleus Assay 2014/1313074

The bone marrow micronucleus test was performed with the test substance in NMRI male mice. Three dose levels of 250, 500 and 1000 mg/kg bw were orally administered once only. Bone marrow smears were obtained from all groups at 24 hours and from the controls and 1000 mg/kg group after 48 hours. Animals given 1000 mg/kg showed signs of reduced activity and ruffled fur.

No increase in the mean frequency of micronucleated polychromatic erythrocytes was observed in the bone marrow of animals treated with the test substance. In the positive control group dosed with cyclophosphamide, the frequency of micronucleated polychromatic erythrocytes showed a highly significant increase.

It was concluded that the test substance did not induce micronuclei in the bone marrow cells of NMRI mice. 

 

Supporting studies: Micronucleus Assay

All studies are GLP and guideline compliant.

In the supporting studies the test substance was also determined to be not mutagenic in the micronucleus assay.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. No mutagenic effects were observed in the in vitro bacterial reverse mutation test, chromosome aberration test, HPRT test and in the in vivo micronucleus assay. As a result the substance is not considered to be classified for mutagenicity under Regulation (EC) No 1272/2008, as amended for the eighth time in Regulation (EU) No 2016/918.