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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Not mutagenic or clastogenic in bacterial and/or mammalian cells in vitro.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Adequate information exists to characterise the mutagenicity of Rosin adduct esters. These are formed after modification of rosin with either fumaric acid or maleic anhydride followed by esterification with glycerol and/or pentaerythritol, and hence the adduct ester products therefore exhibit close structural similarities. The available data includes results of tests conducted using Resin acids and rosin acids, fumarated, esters with pentaerythritol and Rosin, fumarated, reaction products with glycerol and pentaerythritol. This information is summarised below.

Bacterial cell mutation

In a reverse gene mutation assay in bacteria, strains TA 98, TA 100, TA 1535 and TA 1537 of S. typhimurium and strain WP2 of E. coli were exposed to resin acids and rosin acids, fumarated, esters with pentaerythritol in water/ethanol (1:19) at concentrations of 62, 185, 556, 1667 and 5000 µg/plate in the presence and absence of a mammalian metabolic activation system using both the plate incorporation method and the pre-incubation method (Vivotecnia Research, 2010b). No cytotoxicity was observed at any dose level up to the limit concentration of 5000 µg/plate. No experiment with the test substance showed ratios above 2.5 as compared to the negative control, either with or without S9 metabolic activation. No dose response was observed for any of the tested bacterial strains. All vehicle and positive controls induced the appropriate responses in the corresponding strains. Based on the test conditions used in this study, resin acids and rosin acids, fumarated, esters with pentaerythritol was found to be neither mutagenic nor pro-mutagenic.

In a reverse gene mutation assay in bacteria, strains TA 98, TA 100, TA 1535 and TA 1537 of S. typhimurium and strain WP2 of E. coli were exposed to rosin, fumarated, reaction products with glycerol and pentaerythritol in DMSO at concentrations of 60, 190, 560, 1670, 500 µg/plate in the presence and absence of a mammalian metabolic activation system using both the plate incorporation method and the pre-incubation method (Vivotecnia Research, 2010a). No cytotoxicity was observed at any dose level up to the limit concentration of 5000 µg/plate. No experiment with the test substance showed ratios above 2.5 as compared to the negative control, either with or without S9 metabolic activation. No dose response was observed for any of the tested bacterial strains. All vehicle and positive controls induced the appropriate responses in the corresponding strains. Based on the test conditions used in this study, rosin, fumarated, reaction products with glycerol and pentaerythritol was found to be neither mutagenic nor pro-mutagenic.

Mammalian chromosomal aberrations

Resin acids and rosin acids, fumarated, esters with pentaerythritol (CAS No. 94581-15-4), dissolved in ethanol, was assessed for its potential to induce structural chromosomal aberrations in human lymphocytes in vitro in three independent experiments. In each experimental group two parallel cultures were analysed (Harlan Cytotest Cell Research GmbH, 2010). Per culture 100 metaphases were scored for structural chromosomal aberrations, except for the positive control in Experiment II, in the absence of S9 mix, where only 50 metaphases were scored. The highest applied concentration in the pre-test on toxicity (1950.0 µg/mL of the test item) was chosen with regard to the solubility properties of the test item and with respect to the current OECD Guideline 473. Dose selection of the cytogenetic experiment was performed considering the toxicity data and the occurrence of test item precipitation in accordance with OECD Guideline 473. In the absence and presence of S9 mix, concentrations showing clear cytotoxicity were not scorable for cytogenetic damage. In Experiment IB with narrow concentration spacing in the absence of S9 mix and in Experiment II in the absence of S9 mix, clear cytotoxicity was observed at the highest evaluated concentration. In Experiment II in the presence of S9 mix it was not possible to obtain evaluable concentrations in a cytotoxic range. In all independent experiments, neither a statistically significant nor a biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item. No evidence of an increase in polyploid metaphases was noticed after treatment with the test item as compared to the control cultures. Appropriate mutagens were used as positive controls. They induced statistically significant increases (p < 0.05) in cells with structural chromosome aberrations.

Mammalian cell mutation

The study was conducted according to a method that was designed to assess the potential mutagenicity of resin acids and rosin acids, fumarated, esters with pentaerythritol (CAS No. 94581-15-4) on the thymidine kinase, TK +/-, locus of the L5178Y mouse lymphoma cell line (Harlan Laboratories Ltd, 2010c). The method used meets the requirements of the OECD (476) and Method B17 of Commission Regulation (EC) No. 440/2008. Two independent experiments were performed. In Experiment 1, L5178Y TK +/- 3.7.2c mouse lymphoma cells (heterozygous at the thymidine kinase locus) were treated with the test material at eight dose levels, in duplicate, together with vehicle (DMSO) and positive controls using 4-hour exposure groups both in the absence and presence of metabolic activation (2% S9). In Experiment 2, the cells were treated with the test material at eight dose levels using a 4-hour exposure group in the presence of metabolic activation (1% S9) and a 24-hour exposure group in the absence of metabolic activation. The dose range of test material was selected following the results of a preliminary toxicity test. The dose range for Experiment 1 was 10 to 80 µg/ml in the absence of metabolic activation, and 20 to 160 µg/ml in the presence of metabolic activation. The dose range for Experiment 2 was 20 to 160 µg/ml in the absence of metabolic activation, and 40 to 180 µg/ml in the presence of metabolic activation. The maximum dose level used was limited by test material induced toxicity. Precipitate of test material was not observed at any of the dose levels in the mutagenicity test. The vehicle (solvent) controls had acceptable mutant frequency values that were within the normal range for the L5178Y cell line at the TK +/- locus. The positive control materials induced marked increases in the mutant frequency indicating the satisfactory performance of the test and of the activity of the metabolising system. The test material did not induce any toxicologically significant dose-related increases in the mutant frequency at any dose level, either with or without metabolic activation, in either the first or the second experiment. The test material was considered to be non-mutagenic to L5178Y cells under the conditions of the test.

Justification for classification or non-classification

Not classified according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 or UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS).