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Administrative data

Key value for chemical safety assessment

Additional information

Valid experimental data are available to assess the genetic toxicity of Fatty acids, C18-unsatd., dimers, ethoxylated

GENETIC TOXICITY IN VITRO

In vitro:

 

Gene mutation in bacteria:

 

OECD conform studies:

The test substance was tested in two independent tests according to the OECD guideline 471 at a concentration range of 0; 33; 100; 333; 1000; 2750 and 5500 μg/plate (BASF SE, 2012, 40M0072/12M049). The following bacterial strains were used: TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA. The test substance was tested in the absence and pesence of Phenobarbital/ß-Naphthoflavone-induced S9-mix. In both test series the plates were incubated for 48 h at 37°C. According to the results of the present study, the test substance did not lead to a relevant increase in the number of revertant colonies either without S9 mix or after adding a metabolizing system in two experiments carried out independently of each other (standard plate test and preincubation assay). Besides, the results of the negative as well as the positive controls performed in parallel corroborated the validity of this study, since the values fulfilled the acceptance criteria of this study. In this study with and without S9 mix, the number of revertant colonies in the negative controls was within or nearby the range of the historical negative control data for each tester strain. In addition, the positive control substances both with and without S9 mix induced a significant increase in the number of revertant colonies within the range of the historical positive control data or above.

Conclusively, the test item is not a mutagenic test substance in the bacterial reverse mutation test in the absence and the presence of metabolic activation.

 

Gene mutation in mammalian cells:

 

OECD conform studies:

A study was performed to investigate the potential of Fatty acids, C18-unsatd., dimers, ethoxylated to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster according to OECD guideline 476, in two independent experiments (BASF SE, 2013, 50M0072/12X104). 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 concentration used in the range finding pre-experiments was 5.4 µL/mL of the test item or approximately 5.0 µL/mL of the pure substance. The concentration range of the main experiments was limited by precipitation. The test item was dissolved in DMSO. The tested concentrations of the main experiments were 42.5, 84.4, 168.8, 337.5, 675, 1350 µg/mL with and without metabolic activation in the first experiment and without metabolic activation in the second experiment for 4 and 24 hours, respectively. The 4 hour incubation with metabolic activation in the second experiment was done with 84.4, 168.8, 337.5, 675, 1350, 2700 µg/mL. No substantial and reproducible dose dependent increase of the mutation frequency was observed up to the maximum concentration with and without metabolic activation. Appropriate reference mutagens (and DMBA), used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test system 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, Fatty acids, C18-unsatd., dimers, ethoxylated (NIO AD1) is considered to be non-mutagenic in this HPRT assay.

 

Cytogenetic in mammalian cells:

OECD conform studies:

The test item Fatty acids, C 18-unsatd., dimers, ethoxylated, dissolved in DMSO, was assessed for its potential to induce micronuclei in V79 cells of the Chinese hamster in vitro in the absence and presence of metabolic activation by S9 mix according to OECD guideline 487 (BASF SE, 2012, 1483301). Two independent experiments were performed. In Experiment I the exposure period was 4 hours with and without metabolic activation. In Experiment II the exposure period was 24 hours without S9 mix and 4 hours with metabolic activation. The cells were prepared 24 hours (Exp. I and II) after start of treatment with the test item. In each experimental group two parallel cultures were set up and 1000 cells per culture were scored for micronuclei. The highest treatment concentration in this study, 5.4 µL/mL was chosen with regard to the purity (93.6 %) of the test item and with respect to the OECD Guideline for in vitro mammalian cytogenetic tests. In Experiment I, visible precipitation of the test item in the culture medium was observed at 675.0 µg/mL and above in the absence and presence of S9 mix. In addition, precipitation occurred in Experiment II, at 337.5 and 675.0 µg/mL in the absence of S9 mix and at 337.5 µg/mL and above in the presence of S9 mix. Phase separation was observed in Experiment I in a range of 21.1 and 337.5 µg/mL in the absence and presence of S9 mix and at 1350.0 µg/mL and 5.4 µL/mL in the absence of S9 mix. In Experiment II phase separation was observed in a range of 42.2 and 2700.0 µg/mL in the absence of S9 mix and at 168.8 µg/mL and above in the presence of S9 mix. No relevant influence on pH value was observed, osmolarity was slightly decreased in Experiment II at the two highest applied concentrations. In the absence and presence of metabolic activation no cytotoxicity measured as reduced proliferation index was observed. No biologically relevant increase in the percentage of micronucleated cells was observed up to the highest evaluated concentration. The rates of micronucleated cells after treatment with the test item (0.35 - 1.10 %) were close to the solvent control values (0.85 - 1.75 %) and within the range of the laboratory historical solvent control data. In Experiment I in the presence of S9 mix, the value of the solvent control (1.75 %) slightly exceeded the range of the laboratory historical solvent control data (0.15 – 1.50 %). Mitomycin C (0.1 µg/mL), Griseofulvin (9.0 µg/mL) or CPA (15.0 or 20.0 µg/mL) were used as positive controls and showed a distinct increase in the percentage of micronucleated cells.

In conclusion, it can be stated that under the experimental conditions reported, the test item Fatty acids, C 18-unsatd., dimers, ethoxylated did not induce micronuclei in V79 cells (Chinese hamster cell line) in vitro in the absence and presence of metabolic activation. Therefore, Fatty acids, C 18-unsatd., dimers, ethoxylated has to be considered as non-mutagenic in this in vitro test system when tested up to precipitating concentrations.

GENETIC TOXICITY IN VIVO

In vivo cytogenicity:

According to integrated testing strategy, based on available information no further in vivo testing is suggested.

Key study assignment:

For each toxicological endpoint one reliable and suitable study are available. No further reliable and relevant information was available; therefore these studies are integrated as key studies.

Assessment of genetic toxicity:

According to the ITS (guidance on information requirements, R.7.7.6 Integrated Testing Strategy (ITS) for mutagenicity), first in vitro tests were conducted to assess the mutagenic potential. All performed invitro tests assessing the mutagenicity in bactria (BASF SE, 2012, 40M0072/12M049),the gene mutation in mammalian cell (BASF SE, 2013, 50M0072/12X104) and the cytogenetic in mammalian cells (BASF SE, 2012, 1483301) are negative.

As all available in vitro test did not show any positive result and no further hints for mutagenicity can be draw from other information (like structure elements etc.) not further testing is suggested and summarizing no mutagenic effects of the test substance is expected.


Justification for selection of genetic toxicity endpoint
3 GLP and guideline compliant key studies are availabe (Ames test, HPRT and Micronucleus in vitro), all are used according to integrated testing strategy.

Short description of key information:
Gene mutation in bacteria:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA, with and without metabolic activation, OECD 471: negative with and without metabolic activation (BASF SE, 2012, 40M0072/12M049)

Gene mutation in mammalian cells:
HPRT, V79, with and without metabolic activation, OECD 476: non mutagenic with and without metabolic activation (BASF SE, 2013, 50M0072/12X104)

Cytogenetic in mammalian cells:
Micronucleus test in vitro, V79, with and without metabolic activation, two experiments , OECD 487: non mutagenic with and without metabolic activation (BASF SE, 2012, 1483301)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

In summary no hints for mutagenicity obtained from experiments in vitro, therefore the test material does not fulfil the requirement for germ cell mutagens cat. 2 according to GHS (Regulation (EU) 1272/2008) or mutagenic category 3 DPD (67/548/EEC).

 

Labelling mutagenic:

GHS: no labelling

DSD: no labelling