Fatty acids, C18-unsatd., dimers, polymers with 2-ethylhexanol and neopentyl glycol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Genetic toxicity (mutagenicity) in bacteria: negative (RA CAS 147256-33-5, CAS 68440-06-2 and CAS 61788-89-4)

Genetic toxicity (cytogenicity) in mammalian cells: negative (RA CAS 147256-33-5 and CAS 68440-06-2)

Genetic toxicity (mutagenicity) in mammalian cells: negative (RA CAS 68440-06-2 and CAS 61788-89-4)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for read-across

There are no available data assessing genetic toxicity of Fatty acids, C18-unsatd., dimers, polymers with 2-ethylhexanol and neopentyl glycol (CAS 68552-19-2). The assessment of genetic toxicity was therefore based on studies conducted with the source substances Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5), Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters (CAS 68440-06-2) and Fatty acids, C18-unsatd., dimers (CAS 61788-89-4) as part of a read across approach, which is in accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5. in order to fulfil the standard data requirements defined in Regulation (EC) No 1907/2006, Annex VII and VIII, 8.4. Structural similarities and comparable toxicokinetic properties of the source and target substance are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

 

Genetic toxicity (mutagenicity) in bacteria in vitro

CAS 147256-33-5

A reliable gene mutation study in bacteria (Ames test) performed according to OECD TG 471 and in compliance with GLP with Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5) is available (Harlan CCR, 2012). 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 3 to 5000 µg/plate. The test material did not induce cytotoxicity in any of the tested strains at any tested concentration. In the first experiment, precipitation was observed at concentrations ≥333 µg/plate or ≥1000 µg/plate with or without metabolic activation, respectively. In the second experiment precipitation was evident at ≥1000 µg/plate in the presence or absence of S9-mix. Appropriate solvent (acetone) and positive controls were included and gave the expected results. 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.

CAS 68440-06-2

A reliable gene mutation study in bacteria (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 (Harlan Laboratories, 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.

CAS 61788-89-4

A reliable Ames test was performed with the read across substance fatty acids, C18 unsatd., dimers (CAS No. 61788-89-4) equivalent to OECD guideline 471 in S. typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 with and without metabolic activation by rat liver S9-mix (Environmental Safety Laboratory, 1993). The cells were treated with 50, 150, 500, 1500, 5000 µg and 1000, 2000, 3000, 4000, 5000 µg of the test substance/plate. No cytotoxicity was observed and as some significant increases seen in revertant colonies were irreproducible, it is concluded that under the conditions of this study fatty acids, C18 unsatd., dimers did not induce mutations in Salmonella typhimurium strains TA1535, TA1537, TA100 and TA98.

Genetic toxicity (cytogenicity) in mammalian cells in vitro

CAS 147256-33-5

An in vitro mammalian chromosome aberration test according to OECD Guideline 473 was performed with Fatty acids, C18-unsatd., dimers, mixed esters with oleic acid and trimethylolpropane (CAS 147256-33-5) in Chinese hamster lung fibroblasts (V79) (LPT, 2009). In this GLP-conform study, duplicate cell cultures were treated with the test substance at concentrations ranging from 312.5 to 5000 µg/mL for a period of 4 and 20 h (without metabolic activation) or 4 h only (with metabolic activation). These concentrations were selected based on a preliminary cytotoxicity test, in which no toxicity was observed up to the highest concentration of 5000 µg/mL. Consistent with this, treatment of cells in the main study did not induce cytotoxicity in V79 cells up to the maximum concentration of 5000 µg/mL. Based on these results, the analysis of chromosome aberrations was performed at concentrations of 625, 1250, 2500 and 5000 µg/mL, both in the presence or absence of metabolic activation. No significant increase in the number of phases with aberrations at any preparation time and concentration was observed. The positive controls significantly increased the rate of chromosome aberrations, thus indicating the sensitivity of the assay. In conclusion, the test substance was not clastogenic in Chinese hamster lung fibroblasts (V79), neither in the presence nor in the absence of a metabolic activation system, under the chosen experimental conditions.

CAS 68440-06-2

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 (Harlan Laboratories, 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

CAS 68440-06-2

An in vitro gene mutation test in Mouse lymphoma L5178Y cells was performed with Fatty acids, C18-unsatd., dimers, hydrogenated, 2-ethylhexyl esters (CAS 68440-06-2) according to OECD TG 476 and in compliance with GLP (Harlan Laboratories, 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.

CAS 61788-89-4

A mammalian cell gene mutation assay with mouse lymphoma L5178Y cells was performed with the read across substance fatty acids, C18 unsatd., dimers (CAS 61788-89-4) according to OECD guideline 476 (Huntington Research Center, 1993). The cells were treated with 25, 50, 100, 150, 225, 250, 275 and 300 µg/mL with and without metabolic activation for 3 h. After an expression time of 48h and a selection time of 12 days, survival and mutant frequency were determined. The test substance was cytotoxic with (≥ 150 µg/mL) and without metabolic activation (≥300 µg/mL) but showed no influence on the mutation rate of mouse lymphoma L5178Y cells.

 

Conclusion for genetic toxicity

The available data show that the source substances are not mutagenic to bacteria and not mutagenic to mouse lymphoma L5178Y cells in vitro. In addition, the available data with the source substances revealed no clastogenic properties in Chinese hamster lung fibroblasts and in cultured peripheral human lymphocytes in vitro.

Justification for classification or non-classification

Based on the analogue read-across approach, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.