Acetic acid, anhydride, reaction products with 1,5,10-trimethyl-1,5,9-cyclododecatriene

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The substance is negative in the Ames test (OECD TG 471)

The substance is negative in the in vitro Micronucleus Test in Human Lymphocytes (OECD TG 487)

The substance is negative in the in vitro mammalian cell mutation assay using mouse lymphoma L5178Y cells (OECD TG 476).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

OECD TG 471

The substance is tested in the Ames test in accordance with OECD TG 471 and GLP. Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and a tryptophan-dependent mutant of Escherichia coli, strain WP2 uvrA (pKM101), were treated with the test item using both the Ames plate incorporation and pre-incubation methods at up to seven dose levels, in triplicate, both with and without the addition of S9-mix. Based on results of experiment 1 (plate incorporation), the dose range used for experiment 2 (pre-incubation method) was between 5 to 5000 μg/plate. No evidence of mutagenic activity was seen at any concentration of the substance in either mutation test. Adequate negative and positive controls were included. The concurrent positive controls demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations. There were no significant increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) for both experiments. Based on the results, the substance is not mutagenic in this test.

OECD TG 487

The substance is tested in the in vitro Micronucleus Test in Human Lymphocytes in accordance with OECD TG 487 and GLP. Duplicate cultures of human lymphocytes, treated with different concentrations of the test item, were evaluated for micronuclei in binucleate cells at up to four dose levels, together with vehicle (quadruplicate cultures) and positive controls (duplicate cultures). Three exposure conditions were used for the study using a 4-hour exposure in the presence (0, 5, 10, 20, 40, 60, 80, 160 µg/mL Trimofix) and absence (0, 10, 20, 30, 40, 50, 60, 70, 80, 160 µg/mL Trimofix) of a standard metabolising system (S9) at a 2% final concentration, and a 24-hour exposure in the absence (0, 5, 10, 20, 30, 40, 60, 80 µg/mL Trimofix) of metabolic activation. At the end of the exposure period, the cell cultures were washed and then incubated for a further 24 hours in the presence of Cytochalasin B. The substance was tested up into cytotoxic concentrations, but did not induce the formation of micronuclei, with or without metabolic activation. The negative vehicle and positive controls performed as expected. Based on these results, the substance is not cytogenic in this test.

 OECD TG 476

The substance is tested in the in vitro mammalian cell mutation assay using mouse lymphoma L5178Y cells in accordance with OECD TG 476 and GLP. Concentrations up to 2469.6 µg/mL in the presence and absence of metabolic activation were tested. The substance was found to be soluble at approximately 245.17 mg/mL in DMSO. The study consisted of a preliminary toxicity test and two main tests comprising three independent mutagenicity assays. In the preliminary toxicity test, toxicity was observed following a 3-hour exposure to the substance in both the absence and presence of S9 mix and following a 24-hour exposure in the absence of S9 mix. At concentrations from 4.8 to 2469 µg/mL, relative suspension growth (RSG) was reduced from 100 to 0%, from 112 to 0% and from 89 to 0% respectively. Following 3-hour treatment in the absence and presence of S9 mix, the substance did not increase the mutation frequency such that the evaluation criteria for rating the test substance as positive were met. Vehicle and positive controls performed as expected and were comparable with historical control data of this laboratory. In the 24-hour treatment, no increase in the mean mutant frequencies of any of the test concentrations assessed exceeded the sum of the mean concurrent solvent control and the GEF, however, an acceptable reduction in RTG was not obtained (maximum reduction of 33%). Therefore, a further 24-hour treatment was undertaken. Again, no increase in the mean mutant frequencies of any of the test concentrations assessed exceeded the sum of the mean concurrent solvent control and the GEF, within acceptable levels of toxicity. The maximum concentration assessed in this treatment was 48 µg/mL, with the RTG reduced to 13%. Based on the results, the substance not mutagenic in this test.

Justification for classification or non-classification

Based on the results of the gene mutations in bacterial cells (Ames test), cytogenicity information (in vitro mammalian cell mutation assay using mouse lymphoma L5178Y cells) and the gene mutations in mammalian cells (in vitro micronucleus test in human lymphocytes), the substance is not mutagenic and not cytogenic. Therefore it does not have to be classified for genotoxicity in accordance with EU CLP (EC no. 1272/2008 and its amendments).