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Key value for chemical safety assessment

Additional information

In vitro:

In a GLP compliant Ames test, performed according to EU-Method B.14, 5 Salmonella typhimurium strains (TA 1535, TA 1537, TA 1538, TA 98, and TA 100) were used to the test the mutagenic potential of the test substance (10, 100, 333.3, 1000, 5000 µg per plate), both with and without metabolic activation (CCR 1990). No cytotoxic effects were observed in the test groups with and without metabolic activation in both experiments. Up to the highest investigated dose, neither a significant and reproducible increase of the number of revertants was found in any strain as compared to the solvent control nor a concentration-dependent enhancement of the revertant number exists. The presence of liver microsomal activation did not influence these findings. The test substance did not show any mutagenic activity in any strain. Therefore, it was concluded that the test substance did not show any mutagenic activity in any strain.

In a GLP-compliant chromosome aberration test, tested according to OECD guideline 473, chinese hamster lung fibroblast (V79) were exposed to the test substance with and without metabolic activation (CCR 1990). Different exposure times (up to 28 hours) and dosage (up to 5 mg/mL) were chosen in this study. The mitotic index was reduced after treatment with the highest concentrations (with and without S9 mix) indicating that the test substance had cytotoxic properties.There was no relevant increase in cells with structural aberrations after treatment with the test article at any fixation interval either without or with metabolic activation by S9 mix. Therefore, it was stated that the test substance did not induce structural chromosome aberrations in the V79 Chinese hamster cell line.

No studies on the mutagenicity in mammalian cells of FAT40400 were available. However, a HPRT gene mutation test is available for the structural analogue FAT 40426.

In a GLP-compliant mammalian cell gene mutation assay, tested according to OECD guideline 476, Chinese hamster V79 cells were exposed to the test substance with and without metabolic activation and the potential to induce mutations at the HPRT locus was assessed (BSL Bioservice 2013). The selection of the concentrations was based on data from the pre-experiments. Experiment I with and without metabolic activation and experiment II with metabolic activation were performed as a 4h short-term exposure assay. Experiment II without metabolic activation was performed as 20h long time exposure assay. The following concentrations were used. Experiment I without metabolic activation: 5, 10, 25, 50, 100, 250, 500, 1000, 2000 and 2500µg/mL; Experiment I with metabolic activation: 5, 10, 25, 50, 100, 250, 500, 1000, and 2500µg/mL; Experiment II without metabolic activation: 10, 25, 50, 100, 200, 400, 600 and 800µg/mL; Experiment II with metabolic activation: 100, 316, 1000, 1250, 1500, 2000, 2400, 2800, and 3000µg/mL. No precipitation was noted in the experiments. Biologically relevant growth inhibition was observed in experiment I and II with and without metabolic activation. In experiment I without metabolic activation the relative growth was 16.5% for the highest concentration (2500 µg/mL) evaluated. The highest biologically relevant concentration evaluated with metabolic activation was 2500 µg/mL with a relative growth of 17.4%. In experiment II without metabolic activation the relative growth was 12.2% for the highest concentration (800 µg/mL) evaluated. The highest concentration evaluated with metabolic activation was 3000 µg/mL with a relative growth of 21.8%. In both experiments no biologically relevant increase of mutants was found after treatment with the test item (with and without metabolic activation). No dose-response relationship was observed. The positive controls showed distinct biologically relevant effects in mutation frequency. In conclusion, the test substance is considered to be non-mutagenic in the HPRT locus using V79 cells of the Chines hamster.

In vivo:

In a GLP-compliant erythrocyte micronucleus test, tested according to OECD guideline 474, 6 NMRI mice per sex were treated once by oral gavage with the test substance (5000 mg/kg bw) dissolved in 1% CMC followed by a 24, 48 and 72 hours post exposure period (CCR 1990). In a pre-experiment 5000 mg/kg bw was estimated to be the maximum tolerated dose. The finding that the mean number of normochromatic erythrocytes was not increased after treatment with the test article as compared to the negative controls, indicated that the test substancehad no cytotoxic properties. In comparison with the corresponding negative controls there was no enhancement in the frequency of the detected micronuclei at any preparation interval after application of the test article and therefore it could be concluded that the test substance did not induce micronuclei in this study.


Short description of key information:
The test substance is considered to be not mutagenic in the in vitro chromosomal aberration test and Ames test. Furthermore, the in vivo erythrocyte micronucleus test did not show any genetic toxicity. The is substance is also considered to be non-mutagenic in the HPRT mutation assay with a structural analogue.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on the available genotoxicity studies, the test substance does not need to be classified for genotoxicity according to the Directive 67/548/EEC and according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008