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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames Test

Under the conditions of the test the test material was not mutagenic to strain TA98, TA100, TA1535, or TA1537 with or without metabolic activation.

Chromosome Aberration

The test material was considered not to induce any statistically significant increases in the frequency of cells with aberrations and, therefore was considered to be non-clastogenic.

Mouse Lymphoma Assay

Under the conditions of the test, the test material did not induce any increases in the mutant frequency at the TK +/- locus in L5178Y cells that exceeded the Global Evaluation Factor (GEF) of 126 x 10^-6, consequently the test material is considered to be non-mutagenic in this assay.

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Ames Test

A study was conducted to investigate the mutagenic potential of the test material. The study was performed to the basic test system used is that described by Ames, McCann, and Yamasaki (1975), under GLP conditions. The study was assigned a reliability score of 2 in line with the principles for assessing data quality as defined by Klimisch et al. (1997).

The test material was diluted with acetone and tested in the histidine-deficient strains of Salmonella typhimurium TA98, TA100, TA1535, and TA1537 at dose levels of 0.01 to 10 mg/plate with and without metabolic activation provided by Aroclor-induced rat liver S-9. The test material appeared to form a stable emulsion with acetone and the dilutions were well dispersed in the top agar; however, after incubation, test material, at > 0.1 mg/plate, was observed on the surface of the top agar. The test material was cytotoxic at the highest doses to all strains. No reproducible increases in mutant frequency were observed. Under the conditions of the test the test material was not mutagenic to strain TA98, TA100, TA1535, or TA1537 with or without metabolic activation.

Chromosome Aberration

The potential of the test material to induce structural chromosomal aberrations was determined in a GLP study which was conducted in accordance with standardised guidelines OECD 473 and Japanese Guidelines for Screening Mutagenicity Testing Of Chemicals. The study was assigned a reliability score of 2 in line with the principles for assessing data quality as defined by Klimisch et al. (1997).

Duplicate cultures of human lymphocytes, treated with the test material, were evaluated for chromosome aberrations at up to four dose levels, together with vehicle and positive controls. In this study, three exposure conditions were investigated; 4 hours exposure in the presence of an induced rat liver homogenate metabolizing system (S9), at a 2 % final concentration with cell harvest after a 20-hour expression period, 4 hours exposure in the absence of metabolic activation (S9) with a 20-hour expression period and a 24-hour exposure in the absence of metabolic activation.

The dose levels used in the Main Experiment were selected using data from the preliminary toxicity test where the results indicated that the maximum concentration should be limited to the lowest precipitating dose level. The dose levels selected were 7.81, 15.63, 31.25, 62.5, 125 and 250 µg/mL or 4 hour exposure with and without S9 and 24-hours without S9.

All vehicle (Minimal Essential Medium - MEM) controls had frequencies of cells with aberrations within the range expected for normal human lymphocytes.

All the positive control materials induced statistically significant increases in the frequency of cells with aberrations. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated. The test material demonstrated some toxicity in the upper dose range of the Preliminary Toxicity Test, however, the maximum dose level for the Main Experiment was limited to the lowest precipitating dose level and consequently no marked toxicity was observed in the Main Experiment. The test material did not induce any statistically significant increases in the frequency of cells with aberrations, using a dose range that included a dose level that was the lowest precipitating dose level.

Under the conditions of the study, the test material was considered not to induce any statistically significant increases in the frequency of cells with aberrations and, therefore was considered to be non-clastogenic.

Mouse Lymphoma Assay

The mutagenic activity of the test material was evaluated in an in vitro mammalian cell gene mutation test with L5178Y mouse lymphoma cells. The GLP study was conducted in accordance with the standardised guidelines OECD 490 and EU Method B.17, US EPA OPPTS 870.5300 and in alignment with the Japanese MITI/MHW guidelines for testing of new chemical substances.

One main Mutagenicity Test was performed and in this main test, L5178Y TK +/- 3.7.2c mouse lymphoma cells (heterozygous at the thymidine kinase locus) were treated with the test material at 8 dose levels in duplicate, together with vehicle (R0 media), and positive controls using 4-hour exposure groups both in the absence and presence of metabolic activation (2 % S9), and a 24-hour exposure group in the absence of metabolic activation. The dose range of test material used in the main test was selected following the results of a preliminary toxicity test. The test material exhibited dose-related toxicity to the cells in each of the three exposure groups of the preliminary toxicity test. The dose levels plated for viability and expression of mutant colonies were 3.91, 7.81, 15.63, 31.25, 62.5 and 125 µg/mL for 4-hour treatment without S9, 4-hour treatment with S9 (2 %) and 24-hour treatment without S9. The maximum dose level used was limited by the presence of precipitate. A cloudy precipitate of test material was observed at and above 62.5 µg/mL at the end of the exposure period in all three exposure groups. The vehicle control cultures had mutant frequency values that were acceptable for the L5178Y cell line at the TK +/- locus. The positive control substances induced marked increases in the mutant frequency, sufficient to indicate the satisfactory performance of the test and of the activity of the metabolizing system. The test material did not induce any toxicologically significant or dose-related (linear-trend) increases in the mutant frequency at any of the dose levels in the main test, in any of the three exposure groups.

Under the conditions of the test, the test material did not induce any increases in the mutant frequency at the TK +/- locus in L5178Y cells that exceeded the Global Evaluation Factor (GEF) of 126 x 10^-6, consequently it is considered to be non-mutagenic in this assay

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to genetic toxicity.