Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation in bacteria: S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538, with and without metabolic activation, OECD 471: negative with and without metabolic activation (BASF SE, 1999, 9901542) for read across substance Amines, tri-C8-10-alkyl

Gene mutation in mammalian cells: HPRT, V79, with and without metabolic activation, OECD 476, up to 70.0 μg/mL: non mutagenic with and without metabolic activation (BASF SE, 2013, 50M0674/12M311) for read across substance Amines, tri-C8-10-alkyl

Cytogenetic in mammalian cells: Micronucleus test in vitro, V79, with and without metabolic activation, three experiments , OECD 487: non mutagenic with and without metabolic activation (BASF SE, 2013, 33M0674/12M331) for read across substance Amines, tri-C8-10-alkyl Endpoint Conclusion: No adverse effect observed (negative)

Additional information

There is no data available for tridodecylamine. A suitable read across candidate for which data is available is Amines, tri-C8 -10 -alkyl (CAS # 68814 -95 -9):

Gene mutation in bacteria:

 

OECD conform studies:

The test item with the batch number 5J167 was tested in the Salmonella typhimurium Reverse Mutation Assay for the induction of reverse mutations in a bacterial test system. The assay was performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 in two independent experiments, both with and without metabolic activation by S9-mix according to OECD 471 (BASF SE, 1999, 9901542).

Solutions of the test substance were prepared in Tween 80/bidist. water and diluted with Tween 80/bidist. water just before use. The following concentrations were tested:

1st and 2nd test: 8, 40, 200, 1000 and 5000 µg/plate

The direct plate incorporation assay was utilized. The following results were obtained:

Toxic effects were not noted. No enhanced revertant rates compared to concurrent negative controls were observed in all tested strains, neither in the presence nor in the absence of metabolic activation.

Therefore the test substance did not induce reverse mutations in the tested strains of Salmonella typhimurium in this bacterial mutagenicity test.

In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test substance did not induce gene mutations in Salmonella typhimurium strains.

Subsequently, Amines, tri-C8-10-alkyl is considered not to be mutagenic in this bacterial mutagenicity test in vitro.

 

Gene mutation in mammalian cells:

 

OECD conform studies:

The substance Amines, tri-C8-10-alkyl 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 (BASF SE, 2013, 50M0674/12M311). Two independent experiments were carried out, both with and without the addition of liver S9 mix from phenobarbital- and β-naphthoflavone induced rats (exogenous metabolic activation). According to an initial range-finding cytotoxicity test for the determination of the experimental doses and taking into account the cytotoxicity actually found in the main experiments, the following doses were tested.

1st Experiment

without S9 mix

0; 6.3; 12.5; 25.0; 50.0 μg/mL

with S9 mix

0; 2.2; 4.4; 8.8; 17.5; 35.0; 70.0 μg/mL

2nd Experiment

without S9 mix

0; 5.0; 10.0; 20.0; 40.0; 80.0; μg/mL

with S9 mix

0; 1.6; 3.1; 6.3; 12.5; 25.0 μg/mL

Following attachment of the cells for 20-24 hours, cells were treated with the test substance for 4 in the absence of metabolic activation and for 4 hours in the presence of metabolic activation. Subsequently, cells were cultured for 6-8 days and then selected in 6-thioguaninecontaining medium for another week. Finally, the colonies of each test group were fixed with methanol, stained with Giemsa and counted. 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 this study, in the 1st and 2nd Experiment, at least the highest concentrations evaluated for gene mutations were clearly cytotoxic in the absence and the presence of metabolic activation. 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 Amines, tri-C8-10- alkyl is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation.

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells. Therefore, Amines, tri-C8-10- alkyl is considered to be non-mutagenic in this HPRT assay.

 

Cytogenetic in mammalian cells:

OECD conform studies:

The substance Amines, tri-C8-10-alkyl was assessed for its potential to induce micronuclei in V79 cells in vitro (clastogenic or aneugenic activity) according to OECD guideline 487 (BASF SE, 2013, 33M0674/12M331). Three independent experiments were carried out with and without the addition of liver S9 mix from induced rats (exogenous metabolic activation).

The test substance was poorly soluble in culture medium. Due to strong test substance precipitation in an initial range-finding test the following concentrations were tested in this study.

1st Experiment

4 hours exposure; 24 hours harvest time; without S9 mix:

0; 31.3; 62.5; 125.0 µg/mL

4 hours exposure, 24 hours harvest time, with S9 mix:

0; 31.3; 62.5; 125.0 µg/mL

2nd Experiment

24 hours exposure, 24 hours harvest time, without S9 mix

0; 50.0; 100.0; 200.0 µg/mL

4 hours exposure, 24 hours harvest time, with S9 mix

0; 50.0; 100.0; 200.0 µg/mL

3rd Experiment

4 hours exposure, 24 hours harvest time, with S9 mix

0; 100.0; 150.0; 200.0; 250.0 and 300.0 μg/mL

A sample of at least 1 000 cells for each culture were analyzed for micronuclei, i.e.

2 000 cells for each test group. In several cases the sample was increased up to 4 000 cells. The vehicle controls gave frequencies of micronucleated cells within our historical negative control data range for V79 cells. Both positive control substances, EMS and cyclophosphamide, led to the expected increase in the number of cells containing micronuclei. In the main experiments, no cytotoxicity indicated by reduced relative increase in cell count or proliferation index was observed up to the highest applied test substance concentrations. On the basis of the results of the present study, the slight increase in the number of cells containing micronuclei in single test groups as well as the statistical significance both observed only in the presence of a metabolizing system, have to be regarded as biologically irrelevant due to missing reproducibility.

Thus, under the experimental conditions described, Amines, tri-C8-10-alkyl is considered not to have a chromosome-damaging (clastogenic) effect nor to induce numerical chromosomal aberrations (aneugenic activity) under in vitro conditions in V79 cells in the absence and the presence of metabolic activation.

Justification for classification or non-classification

In summary no hints for mutagenicity are obtained from experiments in vitro, therefore the test material does not fulfill the requirement for classification according to GHS (Regulation (EU) 1272/2008) or DSD-DPD (67/548/EEC).

 

Labelling mutagenic:

GHS: no labelling

DSD: no labelling