Registration Dossier

Administrative data

Key value for chemical safety assessment

Additional information

Genetic toxicity in bacteria:

 

The substance Tri-2-ethylhexylamine was tested for mutagenicity in the Ames assay according to a protocol similar to OECD TG 471 (adopted 1981) and following the testing protocol published by Ames (1973, 1975). Different concentrations of the substance (dose range: 20 µg- 5000 µg/plate) were tested in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 in the standard plate test with and without metabolic activation. External metabolic activation was provided by adding S9-mix from Aroclor 1254 treated male Sprague-Dawley rats. In deviation to the current version of the OECD TG 471 no strain was used for the detection of oxidising mutagens and cross-linking agents, i.e. neither TA102 nor Escherichia coli WP2 uvrA nor E. coli WP2 uvrA (pKM101) were used. No bacteriotoxic effect (reduced his-background growth) and no increase in the number of his+ revertants was observed either without S-9 mix or after addition of a metabolizing system. According to the results of the present study, the test substance Tri-2-ethylhexylamine is not mutagenic in the Ames test under the experimental conditions chosen here (BASF AG 1985).

 

Genetic toxicity in mammalian cells:

 

The potential of Tris-(2-ethylhexyl)amin O 2446 to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster was investigated in a study according to OECD TG 476 and GLP regulations by Harlan Cytotest Cell Research GmbH for BASF SE (BASF SE 2010).

The assay was performed in two independent experiments, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 hours. The second experiment was performed with a treatment time of 4 hours with and 24 hours without metabolic activation. The highest applied concentration (3600 µg/mL) was equal to a molar concentration of approximately 10 mM.

No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments. Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test item and the activity of the metabolic activation system.

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. Therefore, Tris-(2-ethylhexyl)amin O 2446 is considered to be non-mutagenic in this HPRT assay.

 

 

In addition, the test item Tris-(2-ethylhexyl)amin O 2446, diluted in tetrahydrofuran (THF), was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in two independent experiments. The assays were conducted in accordance with OECD TG 473 and GLP regulations by Harlan Cytotest Cell Research GmbH for BASF SE (BASF SE 2010).

In the first experiment exposure periods of 4 hours with and without metabolic activation were employed. The preparation interval was 18 hours. In the second experiment exposure periods were 18 and 28 hours without metabolic activation, and 4 hours with metabolic activation, respectively. Accordingly, the preparation intervals were 18 and 28 hours without, and 28 hours with metabolic activation. Metabolic activation was provided by adding S-9 mix from Phenobarbital /ß-naphthoflavone induced male Wistar HsdCpb:WU rat livers.

In each experimental group two parallel cultures were set up. At least 100 metaphases per culture were evaluated for structural chromosome aberrations, except for the positive control in Experiment I without metabolic activation, where only 50 metaphases were evaluated. The highest applied concentration (5µL/mL) was chosen with respect to the current OECD Guideline 473. Dose selection for the cytogenetic experiments was performed considering the toxicity data.

 

In this study in the absence and presence of S9 mix neither test item precipitation nor cytotoxicity was observed up to the highest applied concentration. Phase separation occurred in Experiment I in the presence and absence of metabolic activation.

In both independent experiments no clastogenicity was observed at the concentrations evaluated for cytogenetic damage either with or without metabolic activation. However, two statistically significant increases in the number of aberrant cells excluding gaps were observed after treatment with 0.02 µL/mL (3.5 % aberrant cells excluding gaps) in Experiment I in the presence of S9 mix and with 2.5 µL/mL (3.0 % aberrant cells excluding gaps) in Experiment II in the presence of S9 mix. These observations have to be regarded as biologically irrelevant, since the values were in the range of the laboratory’s historical solvent control data (Exp. I, with S9 mix: 0.0 – 4.0 % aberrant cells excluding gaps; Exp. II, with S9 mix: 0.5 – 3.0 % aberrant cells excluding gaps) and no dose-related increases were observed. No relevant increase in polyploid metaphases was found after treatment with the test item as compared to the frequencies of the control cultures.

Appropriate mutagens were used as positive controls. They induced statistically significant increases (p < 0.05) in cells with structural chromosome aberrations.

In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce structural chromosome aberrations in V79 cells (Chinese hamster cell line) in vitro.

Therefore, Tris-(2-ethylhexyl)amin O 2446 is considered to be non-clastogenic in this chromosome aberration test in the absence and presence of metabolic activation, when tested up to the highest required concentration.

 

 

Genetic toxicity in mammals:

No data available.


Short description of key information:
In vitro studies:
- Ames assay: negative (comparable to OECD TG 471; S. typhimurium strains TA98, TA100, TA1535 and TA1537, with and without metabolic activation)
- HPRT assay: negative (OECD TG 476; V79 cells of the Chinese hamster, with and without metabolic activation)
- Chromosome aberration assay: negative (OECD TG 473; V79 cells of the Chinese hamster, with and without metabolic activation)

In vivo studies: no data available

Endpoint Conclusion:

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data are reliable and suitable for the purpose of classification under Directive 67/548/EEC. Based on the data, classification for genetic toxicity is not warranted under Directive 67/548/EEC.

 

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

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