Amines, C11-14-branched alkyl, monohexyl and dihexyl phosphates

Endpoint summary

Additional information

Ames Test

In a bacterial reverse mutation assay, conducted similar to OECD Guideline 471 the test substance was evaluated for its mutagenic potential based on the ability to induce point mutations in selected loci of Salmonella typhimurium TA 1535, TA 1537, TA 98 and TA 100 (CIBA-Geigy, 1984). The following dose levels were evaluated at indicated condition:

- 1st and 2nd Experiment: 0, 10, 40, 160, 640, 2560 μg/plate (plate incorporation test with and without S9 mix, all strains);

- 3rd Experiment: 0, 1, 4, 16, 64, 256 μg/plate (plate incorporation test without S9 mix, all strains)

Acetone was used as vehicle control and appropriate positive control substances were also evaluated. At concentrations of 256 µg/0.1 ml and above the substance precipitated in soft agar. An inhibiting effect (as reduction in the colony count) was observed at concentrations of 40 and 160 µg/plate without metabolic activation. This effect was more pronounced in the experiment without metabolic activation than in those with activation.

No dose-related biologically relevant increase or doubling in the number of his+ revertants was observed in plate incorporation test in any of the tested strains with or without metabolic activation. According to the results of the present study, the test substance is not mutagenic in the Salmonella typhimurium mutation assay under the experimental conditions chosen. The positive controls yielded the expected results.

HPRT Test

In a GLP-compliant study performed according to OECD guideline 476, the potential of the test item to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster was investigated (Harlan 2013). The assay was performed in two independent experiments, using two parallel cultures each. In the presence of metabolic activation strong toxic effects in one of the parallel cultures were observed in both main experiments at 40.0 µg/mL. No relevant and reproducible increase in mutant colony numbers/10^6 cells was observed in the main experiments up to the maximum concentration with and without metabolic activation. In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

Chromosome Aberration Study

In a GLP compliant study performed according OECD guideline 473 the test item was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro. (Harlan CCR 2013) Two independent experiments were performed. In both experiments, in the absence and presence of S9 mix, no biologically relevant increase in the number of cells carrying structural chromosome aberrations was observed. No biologically relevant increase in the rate of polyploid metaphases was found after treatment with the test item (1.5 - 3.9 %) as compared to the rates of the solvent controls (1.7 - 3.9 %). No biologically relevant increase in the rate of endomitotic metaphases was found after treatment with the test item (0.0 - 0.1 %) as compared to the rates of the solvent controls (0.0 - 0.1 %).No visible precipitation of the test item in the culture medium was observed. In Experiment I in the absence of S9 mix and in Experiment II in the absence of S9 mix with 18 hours continuous treatment concentrations showing clear cytotoxicity were not scorable for cytogenetic damage. A very steep dose-cytotoxicity course was observed in these two experimental parts. In Experiment I and II in the presence of S9 mix and in Experiment II in the absence of S9 mix with 28 hours continuous treatment clear cytotoxicity indicated as reduced mitotic index was observed at the highest evaluated concentration (48.5, 43.4 and 47.1 %, respectively).

Short description of key information:
The material was tested in an Ames test, an HPRT assay and a chromosome aberration test. All assays (Ames test is a pre-GLP test) were performed according to the GLP and OECD guidelines. The test substance was no considered to be mutagenic in bacteria and mammalian cells and was not clastogenic when tested in mammalian cells.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data are considered reliable and suitable for the purpose of classification. Based on the criteria for classification of Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC, classification for genetic toxicity is not warranted.

 

Classifiation, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification. Based on the criteria laid down in Regulation (EC) No. 1272/2008, as amended for the 2nd time in Directive EC 286/2011, classification for genetic toxicity is not warranted.