Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine

Endpoint summary

Additional information

in vitro

Ames:

The test substance Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine was tested for its mutagenic potential based on the ability to induce point mutations in selected loci of several bacterial strains, i.e. Salmonella typhimurium and Escherichia coli, in a reverse mutation assay according to OECD 471 guideline and GLP (BASF, 2013).

STRAINS: TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA

DOSE RANGE: 33 μg – 5 000 μg/plate (SPT)

10 μg – 2 500 μg/plate (PIT; Salmonella strains)

33 μg – 5 000 μg/plate (PIT; E. coli WP2 uvrA)

TEST CONDITIONS: Standard plate test (SPT) and preincubation test (PIT) both with and without metabolic activation (liver S9 mix from induced rats).

SOLUBILITY: Precipitation of the test substance was found from about 2 500 μg/plate onward with and without S9 mix.

TOXICITY: A bacteriotoxic effect was observed depending on the strain and test conditions from about 333 μg/plate onward.

MUTAGENICITY:

A relevant increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S9 mix or after the addition of a metabolizing system.

CONCLUSION:

Thus, under the experimental conditions of this study, the test substance Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine is not mutagenic in the Salmonella typhimurium/Escherichia coli reverse mutation assay in the absence and the presence of

metabolic activation.

HPRT:

The study was performed to investigate the potential of Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster according to OECD 476 guideline and GLP (Harlan, 2012). 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-experiments was 3060 μg/mL based on the solubility properties of the test item. The test item was dissolved in acetone. The concentration range of the main experiments was limited by cytotoxic effects and precipitation. 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 (EMS 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, Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine is considered to be non-mutagenic in this HPRT assay.

Chromosome aberration:

The test item Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine, dissolved in acetone, was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in three independent experiments according to OECD 473 guideline and GLP (Harlan, 2012).

In each experimental group two parallel cultures were set up. 100 metaphases per culture were evaluated for structural chromosome aberrations, except for the positive control in Experiment II after 28 hours continuous treatment without metabolic activation, where only 50 metaphases were evaluated.

The highest applied concentration (3060.0 ug/mL) was chosen with regard to the solubility properties of the test item and with respect to the current OECD Guideline 473.

Dose selection for the cytogenetic experiments was performed considering the toxicity data and the occurrence of test item precipitation.

In Experiment IA in the absence and presence of S9 mix, in Experiment IB in the absence of S9 mix and in Experiment II after 28 hours continuous treatment in the absence of S9 mix and after 4 hours pulse treatment in the presence of S9 mix concentrations showing clear cytotoxicity (reduced mitotic index, cell number or reduced number of metaphases) were not

evaluable for cytogenetic damage. In Experiment IB in the presence of S9 mix no clear cytotoxicity was observed. In Experiment II after 18 hours continuous treatment a markedly reduced mitotic index was observed at the highest evaluated concentration (53.2 % of

control).

No clastogenicity was observed at the concentrations evaluated either with or without metabolic activation.

No evidence of an increase in polyploid metaphases was found after treatment with the test item as compared to the frequencies of the control cultures.

Appropriate mutagens (EMS and CPA) were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations.

In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce structural chromosome aberrations in V79 cells (Chinese hamster cell line) in vitro.

Therefore, Oleic acid, compound with N-(2-aminoethyl)ethane-1,2-diamine is considered to be non-clastogenic in this chromosome aberration test in the absence and presence of metabolic activation, when tested up to cytotoxic or precipitating or the highest evaluable concentrations.

Short description of key information:
Ames-Test (BASF SE, 2013): negative
HPRT test (Harlan, 2012): negative
Chromosome Aberration (Harlan, 2012): negative

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on the negative data received from the mutagenicity tests, no classification according to EU Directive 67/548/EEC and EU classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No 1272/2008 is warranted.