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Key value for chemical safety assessment

Additional information

Ames-Test:

Five bacterial strains, Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were used to investigate the mutagenic potential of the test item in two independent experiments in accordance with OECD guideline No. 471. Each assay was conducted with and without metabolic activation (S9 Mix). The following test item concentrations were used: 8 -500 µg of test item per plate.In the performed experiments positive and negative (vehicle) controls were run concurrently. The findings show that Di-n-octylcarbonat did not induce any reverse mutations in the absence or on the presence of S9 -mix-induced solutions. The validity criteria of the study were fulfilled. The reported data of this mutagenicity assay shows, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the strains used. Therefore, the test item is considered non-mutagenic in this bacterial reverse mutation assay.

Chromosome aberration test:

The test item dissolved in ethanol was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in the absence and presence of metabolic activation by S9 mix according to OCED guideline No. 473 and EU Method B.10.

Two independent experiments were performed. The chromosomes were prepared 18 h and 28 h after start of treatment with the test item. In each experimental group two parallel cultures were set up. Per culture 100 metaphase per plate were scored for structural chromosome aberrations. In both independent experiments, neither a significant nor a biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item. No increase in the frequencies of polyploid metaphases was found after treatment with the test item as compared to the frequencies of the controls.

Appropriate mutagens were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations. Under the experimental conditions reported, the test item did not induce structural chromosome aberrations as determined by the chromosome aberration test in V79 cells (Chinese hamster cell line) in vitro. Therefore, the test item is considered to be non-clastogenic in this chromosome aberration test.

in vitro gene mutation in mammalian cells (HPRT assay)

The study was performed to investigate the potential to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster.

The assay was performed in two independent experiments, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation.

The highest concentration applied in the pre-experiment was3000 µg/mL (equal to 10 mM). The concentration range of the main experiments was limited by the solubility of the test item in aqueous medium and cytotoxic effects.The test item was dissolved in ethanol.

The tested concentrations of the main experiments are described in chapter12.6(page 18). The evaluated experimental points and the results are summarised in table 1 (page 13).

No substantial and reproducible dose dependent increase of the mutation frequency was observed up to the maximum concentration with and without metabolic activation.

Appropriate reference mutagens (and DMBA), used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test system and the activity of the metabolic activation system.

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, the substance is considered to be non-mutagenic in this HPRT assay.


Short description of key information:
Ames test:
The test item was tested for mutagenic activity in a bacterial reverse mutation assay with five strains of S. typhimurium with and without metabolic activation. The reported data of this mutagenicity assay shows, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the strains used. Therefore, the test item is considered non-mutagenic in this bacterial reverse mutation assay.

Chromosome aberration test:
The test item was tested for structural chromosome aberrations in V79 cells (Chinese hamster cell line ). The reported data shows, that under the experimental conditions reported, the test item did not induce structural chromosome aberrations.

HPRT test
The test substance is not considered to be mutagenic in the HPRT test.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on the available in genotoxicity studies, there is no indication for a mutagenic/genotoxic potential of the test substance. Thus, the test substance has not to be classified with regard to genotoxicity according to Directive 67/548/EEC and to Regulation (EC) No 1272/2008 (CLP).