Fatty acids, C18-unsatd, dimers, hydrogenated, diisopropyl esters

Endpoint summary

Additional information

Justification for grouping of substances and read-across

There are only limited data available on genetic toxicity of Fatty acids, C18-unsatd., dimers, hydrogenated, diisopropyl esters (CAS 103213-20-3). In order to fulfil the standard information requirements set out in Annex VII and VIII, 8.4., in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, 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 of genetic toxicity :

CAS	Chemical name	Molecular weight	Gene mutation in vitro (Ames)	Chromosome aberration in vitro	Gene mutation in vitro (MLA)
103213-20-3 (a)	Fatty acids, C18-unsatd., dimers, hydrogenated, diisopropyl esters	645	RA: CAS 68440-06-2	RA: CAS 68440-06-2	RA: CAS 68440-06-2
68440-06-2 (b)	Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters	789	Experimental result: negative	Experimental result: negative	Experimental result: negative

(a) The substance subject to registration is indicated in bold font.

(b) Reference (read-across) substances are indicated in normal font. Lack of data for a given endpoint is indicated by “--“.

The above mentioned substances are considered to be similar on the basis of the structural similar properties and/or activities. The available endpoint information is used to predict the same endpoints for Fatty acids, C18-unsatd., dimers, hydrogenated, diisopropyl esters (CAS 103213-20-3). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Discussion

No data on genetic toxicity is available with Fatty acids, C18-unsatd., dimers, hydrogenated, diisopropyl esters (CAS 103213-20-3). Therefore, read across from the structurally analogue Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters (CAS 68440-06-2) was applied.

Genetic toxicity (mutagenicity) in bacteria in vitro

A reliable bacterial gene mutation study (Ames test) performed according to OECD TG 471 and in compliance with GLP with Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters (CAS 68440-06-2) is available (Wisher, 2015). The strains Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and E.coli WP2 uvr A were tested according to the plate incorporation (experiment I) and pre-incubation (experiment II) procedure in the absence and presence of a metabolic activation system (phenobarbitone/β-naphthoflavone-induced rat liver S9-mix). Two independent experiments were conducted in three repetitions at concentrations from 15 to 5000 µg/plate (experiment I) and 50 to 5000 µg/plate (experiment II). The test material did not induce cytotoxicity in any of the tested strains at any tested concentration. Appropriate solvent (acetone) and positive controls were included and gave the expected results. Precipitation of the test material was recorded at concentrations ≥1500 µg/plate in both experiments (with and without metabolic activation). No significant increase in the number of revertants was observed in any of the tester strains with and without metabolic activation. Based on the results of the study the test material was considered to be not mutagenic to bacteria under the conditions of the test.

 

Genetic toxicity (cytogenicity) in mammalian cells in vitro

An in vitro chromosome aberration test in human lymphocytes was performed with Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters (CAS 68440-06-2) according to OECD TG 473 and in compliance with GLP (Bowles, 2015). Human peripheral lymphocytes were cultured and treated with the test material or vehicle (acetone) in the absence or presence of a metabolic activation system (phenobarbitone/β-naphthoflavone-induced rat liver S9-mix) in duplicates at concentrations of 2.5, 5, 10, 20, 40 and 60 µg/mL for 4 or 24 hours (without S9-mix) and 4 hours (with 2% S9-mix). Fixation and staining of the cells were performed 24 hours after start of exposure with the test material. Cytotoxicity was assessed by determination of the mitotic index. Appropriate solvent and positive controls were included in the test and gave the expected results. The test substance did not cause a statistically significant, dose-related increase in chromosome aberrations under the tested conditions. Slight toxicity was only observed at 40 µg/mL in the 24-hour continuous exposure group (without S9-mix). Precipitation of the test material was recorded at 60 µg/mL in the 4-hour exposure group in the presence of S9-mix, and at and above 40 µg/mL in the 4 and 24-hour exposure group in the absence of S9-mix. Based on the results of the study the test material is considered not to be clastogenic in this chromosome aberration test in vitro.

 

Genetic toxicity (mutagenicity) in mammalian cells in vitro

An in vitro gene mutation test in Mouse lymphoma L5178Y cells was performed Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters (CAS 68440-06-2) according to OECD TG 476 and in compliance with GLP (Brown, 2015). Mouse lymphoma cells were treated with the test material or vehicle (acetone) in the absence or presence of a metabolic activation system (phenobarbitone/β-naphthoflavone-induced rat liver S9-mix) in duplicates at concentrations from 4.88 to 156.25 µg/mL for 4 hours (+S9-mix: experiment I + II and -S9-mix: experiment I) and 9.77 to 312.5 µg/mL for 24 hours (-S9-mix: experiment II). After exposure with the test material mouse lymphoma cells were cultured in microtiter plates containing trifluorothymidine (4 µg/mL) selective medium for additional 10 - 14 days. Appropriate solvent and positive controls were included in the test and gave the expected result. No evidence of marked toxicity following exposure to the test item in either the absence or presence of metabolic activation was recorded in both experiments. A cloudy precipitate of the test item was observed in both experiments at and above 39.06 µg/mL which turned to a greasy/oily precipitate at 156.25 µg/mL at the end of exposure. The test substance did not cause a statistically significant, dose-related increase in mutant frequency under the tested conditions. Based on the results of the study the test material is considered not to be mutagenic in this mouse lymphoma test in vitro.

Justification for selection of genetic toxicity endpoint
No study was selected, since all available in vitro genetic toxicity studies were negative.

Short description of key information:
In vitro gene mutation:
Bacterial reverse mutation assay (Ames test / OECD 471): negative (RA CAS 68440-06-2)
In vitro chromosome aberration test in human lymphocytes (CA / OECD 473): negative (RA CAS 68440-06-2)
In vitro gene mutation assay in mouse lymphoma cells (MLA / OECD 476): negative (RA CAS 68440-06-2)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The available data on genetic toxicity of Fatty acids, C18-unsatd., dimers, hydrogenated, diisopropyl esters do not meet the criteria for classification according to Regulation (EC) 1272/2008 and are therefore conclusive but not sufficient for classification.