Ethyl linoleate

Endpoint summary

Additional information

Bacterial Reverse Mutation Assay (Ames Test)

A GLP compliant bacterial reverse mutation test was performed according to OECD 471 and EU method B.13/14 (BASF, 2010). 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 independent experiments in the presence and absence of S9 mix (rat liver S9 mix induced by a combination of phenobarbital and beta-naphthoflavone). The test substance was dissolved in ethanol. In the dose range finding test, the test substance was tested up to concentrations of 5000 µg/plate in the absence and presence of S9 mix in the strains TA100 and WP2uvrA. the test substance precipitated on the plates at dose levels of 1000 µg/plate and upwards. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed. Results of this dose range finding test were reported as part of the first experiment of the mutation assay.

Based on the results of the dose range finding test, the test substance was tested in the first mutation assay at a concentration range of 10 to 1000 µg/plate in the absence and presence of 5% (v/v) S9 mix in tester strains TA1535, TA1537 and TA98. In an independent repeat of the assay with additional parameters, the test substance was tested at the same concentration range as the first assay in the absence and presence of 10% (v/v) S9 mix in tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA. the test substance precipitated on the plates at the top dose of 1000 µg/plate. The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed. The test substance did not induce a significant dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9 metabolic activation. These results were confirmed in an independently repeated experiment.

In this study, the negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.

Based on the results of this study it is concluded that the test substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Chromosome Aberration Assay

A GLP compliant chromosome aberration assay was performed according to OECD 473 and EU method B.10 (BASF, 2010). The possible clastogenicity of the test substance was tested in two independent experiments with cultured peripheral human lymphocytes. In the first cytogenetic assay, the test substance was tested up to 333 µg/mL for a 3 h exposure time with a 24 h fixation time in the absence and presence of 1.8% (v/v) S9-fraction. The test substance precipitated in the culture medium at this dose level. In the second cytogenetic assay, the test substance was tested up to 800 µg/mL for a 24 h continuous exposure time with a 24 h fixation time and up to 600 µg/mL for a 48 h continuous exposure time with a 48 h fixation time in the absence of S9 mix. Appropriate toxicity was reached at these dose levels. In the presence of S9 mix the test substance was tested up to 333 µg/mL for a 3 h exposure time with a 48 h fixation time. The test substance precipitated in the culture medium at this dose level. The number of cells with chromosome aberrations found in the solvent control cultures was within the laboratory historical control data range. Positive control chemicals, mitomycin C and cyclophosphamide, both produced a statistically significant increase in the incidence of cells with chromosome aberrations, indicating that the test conditions were adequate and that the metabolic activation system (S9 mix) functioned properly. The test substance did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and presence of S9 mix, in either of the two independently repeated experiments. No effects of the test substance on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both in the absence and presence of S9 mix. Therefore it can be concluded that the test substance does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions described.

Finally, it is concluded that this test is valid and that the test substance is not clastogenic in human lymphocytes under the experimental conditions described.

Mammalian Cell Gene Mutation Test

A GLP compliant mouse Lymphoma assay was performed according to OECD 476 and EU method B.17 (BASF, 2010). The test substance was suspended in dimethyl sulfoxide. The test was performed in two independent experiments in the absence and presence of S9 mix (rat liver S9 mix induced by a combination of phenobarbital and beta-naphthoflavone).

In the first experiment, the test substance was tested up to concentrations of 150 and 300 µg/mL in the absence and presence of 8% (v/v) S9 mix, respectively. The incubation time was 3 hours. The test substance was tested up to cytotoxic levels of 82 and 88% in the absence and presence of S9 mix, respectively.

In the second experiment, the test substance was tested up to concentrations of 90 and 300 µg/mL in the absence and presence of 12% (v/v) S9 mix, respectively. The incubation times were 24 hours in the absence of S9 mix and 3 hours in the presence of S9 mix. The test substance was tested up to cytotoxic levels of 74 and 86% in the absence and presence of S9 mix, respectively.

The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range and within the acceptability criteria of this assay. Mutation frequencies in cultures treated with positive control chemicals were increased by 11- and 15-fold for MMS in the absence of S9 mix, and by 11- and 13-fold for CP in the presence of S9 mix. It was therefore concluded that the test conditions, both in the absence and presence of S9 mix, were appropriate and that the metabolic activation system (S9 mix) functioned properly. In the absence of S9 mix, the test substance did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent repeat experiment with modifications in the duration of treatment time. In the presence of S9 mix, the test substance did not induce a significant increase in the mutation frequency in the first experiment. This result was confirmed in an independent repeat experiment with modifications in the concentration of the S9 for metabolic activation.

It is concluded that the test substance is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described.

Short description of key information:
RA: 91051-53-5:
Ames Test (BASF, 2010): negative
in vitro chromosome aberration test (BASF, 2010): negative
Mouse Lymphoma Test (BASF, 2010): negative

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available in vitro studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for mutagenicity under Directive 67/548/EEC, as amended for the 31st time in Directive2009/2/EC.

 

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. As a result the substance is not considered to be classified for mutagenicity under Regulation (EC) No 1272/2008, as amended for the fifth time in Directive EC 944/2013.