Fatty acids, C16-18 and C18-unsatd., Et esters

Endpoint summary

Additional information

Justification for grouping of substances and read-across

There are no data available for the genetic toxicity of Fatty acids, C16-18 and C18-unsatd., ethyl esters (CAS 85049-36-1). In order to fulfil the standard information requirements set out in Annex IX, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, a read-across from structurally related substances was conducted.

In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across).

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006 whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Overview for genetic toxicity

CAS #	Genetic toxicity (mutagenicity) in bacteria in-vitro	Genetic toxicity (cytogenicity) in mammalian cells in-vitro	Genetic toxicity (mutagenicity) in mammalian cells in-vitro	Genetic toxicity in vivo
85049-36-1 Target substance	RA: 544-35-4	RA: 544-35-4	RA: 544-35-4	--
544-35-4	negative	negative	negative	--

The above mentioned substance is considered to be similar on the basis of structural similarity resulting in similar properties and/or activities. The available endpoint information is used to predict the same endpoints for Fatty acids, C16-18 and C18-unsatd., ethyl esters (CAS 85049-36-1).

A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

AMES, OECD 471, negative

A bacterial reverse mutation assay was performed with 9,12-Octadecadienoic acid (Z,Z)-,Ethyl Ester (CAS# 544-35-4) according to OECD guideline 471 and GLP with the S. typhimurium strains TA98, TA100, TA1535 and TA1537 and the E. coli strain WPA2uvr A (Verbaan, 2010). The bacterial tester strains were treated with 10, 33, 100, 333, 1000, 3330 and 5000 µg/plate of the test substance in absence and presence of metabolic activation by phenobarbital- and ß-naphthoflavone-induced rat liver S9-mix. Two independent experiments were performed with triplicates each. Sodium Azide, 9-Aminoacridine, 2-Nitrofluorene, Methylmethanesulfonate and 4-Nitroquinoline-N-oxide and 2-Aminoanthracene were used as positive controls without and with metabolic activation, respectively. Positive control materials induced statistically significant increases in the frequency of revertant colonies indicating the satisfactory performance of the test and the activity of the metabolizing system. The test substance slightly precipitated at concentrations ≥ 1000 µg/plate but induced no cytotoxic or genotoxic effects at any concentration neither in the presence nor in the absence of metabolic activation. Based on the study results,

9,12-Octadecadienoic acid (Z,Z)-,Ethyl Ester did not induce gene mutations in four tested Salmonella and one tested E. coli strain.

 

In vitro mammalian chromosome aberration test, OECD 473, negative

The ability of Ethyl Linoleate (CAS# 544 -35 -4) to induce chromosome aberrations in cultured peripheral human lymphocytes was tested according to OECD guideline 473 and under GLP (Verbaan, 2010). Test substance concentrations of up to 800 µg/mL dissolved in DMSO were tested in the presence and absence of metabolic activation. At concentrations of 333 µg/mL and higher precipitation of test substance occurred. The first experiment was incubated for 3 hours and 24 hours fixation with and without metabolic activation, whereas the second experiment was incubated for 3 hours together with 48 hours fixation (with S9 mix) and 24 and 48 hours incubation without S9 mix followed by 24 and 48 hours fixation, respectively. 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. Ethyl Linoleate 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 on the number of polyploid cells and cells with endoreduplicated chromosomes were observed.

 

In vitro mammalian cell gene mutation test, OECD 476, negative

An in vitro Mammalian Cell Gene Mutation Assay according to OECD Guideline 476 and GLP was performed with Ethyl Linoleate (CAS# 544-35-4-13-3) in mouse lymphoma L5178Y cells (Verspeek-Rip, 2010). The cells were treated for 3 and 24 hours with 5% (v/v) and without S9-mix and with 10% (v/v) and without S9-mix, respectively. The test substance was tested up to 300μg/mL dissolved in DMSO. Precipitation was seen at 100µg/mL and higher. At this concentration cytotoxicity occurred in the presence and absence of metabolic activation. Cyclophosphamide and Methylmethanesulfonate were used as positive controls with and without S9 mix, respectively. Positive and negative controls were valid and in range of historical control data. No significant increase in mutation frequency occurred in any of the test conditions, indicating that Ethyl Linoleate is not mutagenic in the mammalian cells in vitro.

Conclusion:

One study assessing the potential genetic toxicity using analogue based read-across from structural related substance 9,12-Octadecadienoic acid (Z,Z)-,Ethyl Ester (CAS 544-35-4) in bacteria (Ames test) is available. In vitro chromosomal aberration test and an in vitro mammalian cell gene mutation assay were also performed with ethyl linoleate (CAS 544-35-4). The results of all the tests were negative. The available data on genetic toxicity in vitro indicates that the structural related substance do not have genetic toxicity potential.

Justification for selection of genetic toxicity endpoint
Hazard assessment is conducted by means of read-across based on an analogue approach. No study was selected since all available in vitro and in vivo genetic toxicity studies were negative. All available studies are adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substance and overall quality assessment (refer to the endpoint discussion for further details).

Short description of key information:
Negative results in several Salmonella typhimurium strains , with and without metabolic activation (OECD 471, GLP, analogue approach).
Negative results in mammalian chromosomal aberration test (OECD 473, GLP, analogue approach).
Negative results in mammalian cell gene mutation tests (OECD 476, GLP, analogue approach).

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on read-across from structurally similar substances, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.