Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

in vitro Gene mutagenicity in bacteria Ames test (S. typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538 and E. coli WP2 uvrA), with or without metabolic activation: negative (OECD 471, BASF 1985). 

Gene mutagenicity in mammalian cells Mouse lymphoma assay, L5178Y cells, with or without metabolic activation: negative (OECD 476; BASF 1996) 

Cytogenicity in mammalian cells Micronucleus formation in vitro, CHL V79, with or without metabolic activation: negative (OECD 487; BASF 2001). 

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Gene mutagenicity in bacteria
A reverse gene mutation assay in bacteria, performed according to OECD guideline 471 is available (BASF 1995). The tester strains used in this assay were Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, TA 1538 and the Escherichia coli tester strain WP2 uvrA. The strains were exposed to concentrations of 100, 250, 500, 1000, 2500 and 5000 µg/plate in the presence and absence of S9 mix along with concurrent vehicle (DMSO) and positive controls. Under the conditions of this test, the test substance did not cause a positive increase in the number of revertants per plate of any of the tester strains either in the presence or absence of S9 mix.
 
Gene mutagenicity in mammalian cells
The target substance was tested for genetic toxicity in vitro in a cell gene mutation test with mouse lymphoma cells according to OECD guideline 476 (CHV 1996). In the mutation assay test substance concentrations (suspension in DMSO) of 3.13, 6.25, 12.5, 25.0, 50.0 and 100 µg/mL were tested in the presence and absence of rat liver S9 metabolic activation. Higher concentrations were not tested because of the insoluble nature of the test substance. All dosing solutions were washed to remove the visible precipitate before application. Two trials of the non-activation and the S9 metabolic activation mutation assay were performed but the first trial was unacceptable because of problems in the cell cultures. In Trial 2, six treatments from 3.13 to 100 µg/mL were initiated and all doses were cloned for mutation analysis. No cytotoxicity was observed under either activation conditions.

None of the six analyzed treatments with or without metabolic activation included a mutation frequency that exceeded the minimum criterion for a positive response. Thus, the test substance was negative with or without metabolic activation in the TK locus in L5178Y lymphoma cells under the conditions of this study.

 
Cytogenicity in mammalian cells

A GLP compliant in vitro micronucleus test was performed according to OECD guideline 487 (BASF 2001). CHL V79 cells were exposed to the following test substance doses:

Mixed Population Method
24 h exposure, 24 h harvest time, without S9-mix
0, 1.25, 2.5, 5, 10, 15 µg/mL

4 h exposure, 24 h harvest time, with S9-mix
0, 1.25, 2.5, 5, 10, 15 µg/mL

Mitotic Shake Off Method (24 h mitotic shake off)
24 h exposure, 27 h preparation time, without S9-mix
0, 0.78, 1.56, 3.125, 6.25, 12.5 µg/mL

4h exposure, 27 h preparation time, with S9-mix
0, 1.56, 3.125, 6.25, 12.5, 25 µg/mL

Strong test substance precipitation in the vehicle was observed at all test doses. In culture the test substance was obviously soluble up to 6.25 µg/mL.

On the basis of the results of the present study, the test substance did not cause any increase in the number of cells containing micronuclei either without S-9 mix or after adding a metabolizing system. These findings were confirmed in two modified versions of the in vitro micronucleus assay carried out independently of each other.
Thus, under the experimental conditions of this assay, the test substance is considered not to be a chromosome-damaging (clastogenic) agent nor did it induce numerical chromosomal aberrations (aneugenic activity) under in vitro conditions in V79 cells .

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. Based on the available in vitro data, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the ninth time in Regulation (EC) No 2016/1179.