Reaction mass of Pentaerythritol bis (2-ethylhexanoate) bis (3,5,5-trimethylhexanoate) and Pentaerythritol tris (2-ethylhexanoate) 3,5,5-trimethylhexanoate and Pentaerythritol 2-ethylhexanoate tris (3,5,5-trimethylhexanoate) and Pentaeryhthritol tetrakis(2-ethylhexanoate) and Pentaerythritol tetrakis (3,5,5-trimethylhexanoate)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test (OECDTG 471): negative

In vitro chromosome aberration test (EU Method): negative

Genemutation in mammalian cells (OECDTG 490): negative

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Ames test:

An Ames test was performed according to OECD 471 guideline and GLP principles.

RB68 was initially tested in the tester strains TA100 and WP2uvrA as a dose-range finding test with concentrations of 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate in the absence and presence of S9-mix. Based on the results of the dose-range finding test, the following dose-range was selected for the mutation assay with the tester strains, TA1535, TA1537 and TA98 in the absence and presence of S9-mix: 52, 164, 512, 1600 and 5000μg/plate.

RB68 precipitated on the plates at the highest dose level only. 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.

To obtain more information about the possible mutagenicity of the test item, a pre-incubation experiment was performed in the absence and presence of S9-mix. Based on the results of the first mutation assay, RB68 was tested up to the dose level of 5000 µg/plate in the tester strains TA1535, TA1537, TA98, TA100 and WP2uvrA. There was no reduction in the bacterial background lawn and no biologically relevant decrease in the number of revertants at any of the concentrations tested in all tester strains in the absence and presence of S9-mix.

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.

All other bacterial strains showed negative responses over the entire dose range, i.e. no significant dose-related increase in the number of revertants in two independently repeated experiments.

In conclusion, based on the results of this study it is concluded that RB68 is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Chromosome aberration test:

In a chromosome aberration study, cultured peripheral human lymphocytes were exposed to different concentrations of RB68, in the presence and absence of S9-mix according to an EU Method and GLP principles. RB68 was tested up to and including the concentration of 5000 μg/mL, with and without S9 -mix. The exposure period was 22 and 46 hours without S9 and 4 hours with S9.

RB68 induced a decrease in the mitotic index both with and without metabolic activation. Reliable negative and positive controls were included.

The test material RB68 demonstrated no significant increases in the frequency of cells with aberrations in any of the treatment conditions. The substance has been shown to be non-clastogenic to human lymphocytes under the conditions of this test.

MLA:

The mouse lymphoma assay with the substance was conducted according to OECD 490 guideline and GLP principles.

In the first experiment, RB68 was tested up to concentrations of 78 μg/mL in the absence and presence of S9-mix, respectively. The incubation time was 3 hours. In the second experiment, RB68 was again tested up to concentrations of 78 μg/mL in the absence of S9-mix. The incubation time was 24 hours. The test item precipitated in the culture medium at this dose level. Reliable negative and positive controls were included. No significant toxicity was observed and all dose levels were evaluated in the absence and presence of S9-mix in both experiments.

In the absence of S9-mix, RB68 did not induce a biologically relevant increase in the mutation frequency in the first experiment. This result was confirmed in an independent experiment with modification in the duration of treatment. In the presence of S9-mix, RB68 did not induce a biologically relevant increase in the mutation frequency.

In conclusion, RB68 is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described in this study.

Justification for classification or non-classification

Based on the results of the Ames test, in vitro chromosome aberration and mouse lymphoma studies, the substance RB68 does not have to be classified for genotoxicity in accordance with Regulation (EC) No 1272/2008 and its updates.