Sodium hydrogen N-(1-oxotetradecyl)-L-glutamate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Different tests were done in order to assess genetic toxicity.

All were negative.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In vitro:

A study was conducted to determine the in vitro genetic toxicity according to OECD Guideline 471 and EU Method B.13/14, in compliance with GLP. Dose range finding tests as well as direct plate and pre-incubation assays both in the absence and presence of S9-mix were performed. Salmonella typhimurium strains TA1535, TA1537, TA100 and TA98 and Escherichia coli strain WP2uvrA were exposed to the test substance at concentration levels of 0.55, 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate, and also exposed to the negative or positive control substances for 48 ± 4 h (plus a pre-incubation time of 30 min if needed). In the dose range finding study, the test substance was initially tested up to 5000 µg/plate in the tester strains TA100 and WP2uvrA through a direct plate assay. The test substance showed precipitation at the highest dose level. Cytotoxicity was evidenced in TA100 in the absence and presence of S9 -mix. In tester strain WP2uvrA, no toxicity was observed at any of the dose levels tested. In the first mutation experiment, the test substance was tested up to concentrations of 1600 and 5000 µg/plate in the absence and presence of S9-mix, respectively in the tester strains TA1535, TA1537 and TA98. The test substance precipitated on the plates at the dose level of 5000 μg/plate. Cytotoxicity was observed in all tester strains in the absence and presence of S9-mix. Since the test substance was severely cytotoxic in the first mutation experiment, an additional dose range finding test was performed with strains TA100 and WP2uvrA, both with and without S9-mix according to the pre-incubation method. In this dose range finding study, the test substance was initially tested up to concentrations of 512 and 5000 µg/plate in the tester strains TA100 and WP2uvrA, respectively. The test substance precipitated at dose levels of 1600 and 5000 μg/plate and therefore the number of revertants of this dose level could not be determined. Cytotoxicity was observed in tester strain TA100 in the absence and presence of S9-mix. In tester strain WP2uvrA, no toxicity was observed up to the dose level of 1600 μg/plate. In the second mutation experiment, the test substance was tested up to concentrations of 164 and 512 µg/plate (in absence and presence of S9-mix, respectively) in the tester strains TA1535, TA1537 and TA98 in the pre-incubation assay. Cytotoxicity was observed in all three tester strains in the absence and presence of S9-mix. The test substance did not induce a significant dose-related increase in the number of revertant (His+) colonies in any of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in tester strain WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in an independently repeated experiment. The negative and strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly. Under the study conditions, the substance was not mutagenic in the Salmonella typhimurium and Escherichia coli reverse mutation assay (Verspeek-Rip, 2006).

A study was conducted to determine the in vitro genetic toxicity of the read-across substance L-glutamic acid, N-coco acyl derivs., monosodium salts according to OECD Guideline 471, in compliance with GLP. Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 were exposed to the test substance at concentration levels of 0, 4, 20, 100, 500, 2500 and 5000 µg/plate with and without metabolic activation (S9-mix induced Aroclor 1254), and to negative or positive control substances for 48 h. In the dose range finding study, the test substance proved to be toxic for the bacterial strain TA 100 in the absence of a metabolizing system at a dose of 5000 µg/plate. Therefore 5000 µg/plate was chosen as the highest dose in the experiment. The test substance did not precipitate on the plates up to the highest investigated doses. The test substance did not induce toxicity or an increase in number of revertants at any concentration level. Under the study conditions, the substance was not mutagenic in S. typhimurium with and without metabolic activation (Hoechst, 1994).

A study was conducted to determine the in vitro genetic toxicity of the read-across substance L-glutamic acid, N-coco acyl derivs., disodium salts according to OECD Guideline 487, in compliance with GLP. Two independent experiments were performed in Chinese hamster lung fibroblasts (V79 cells). Cells were exposed to the test substance (diluted in water) at concentrations from 0 to 1600 µg/mL for either 4 or 24 h with and without metabolic activation (S9 mix).Cytotoxicity, indicated by a clearly reduced proliferation index, cell numbers or a low quality of the slides was observed in the absence of S9 mix at 700 µg/mL (1st experiment) and at 800 µg/mL (2nd and 3rd experiment). In the presence of S9 mix, cytotoxic effects were obtained 800 µg/mL onwards but only in the 2nd experiment. In the 1st experiment a single statistically significant increase in the number of micronucleated cells was observed at 700 µg/mL after 4 h exposure in the absence of metabolic activation. However, this finding was neither confirmed in the repeat experiment under similar conditions (3rd experiment) nor in any additional experiment under modified test conditions (24 h exposure in the absence of metabolic activation or 4 h exposure in the presence of metabolic activation). The finding was considered artifactual due to the strong cytotoxicity in this test group (RICC 41.4%). Under the study conditions, the substance was not clastogenic in Chinese hamster lung fibroblast V79 cells (BASF SE, 2013).

A study was conducted to determine the in vitro genetic toxicity of the read-across substance L-glutamic acid, N-coco acyl derivs., disodium salts according to OECD Guideline 476, in compliance with GLP. The potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus was evaluated in Chinese hamster ovary (CHO) cells in vitro. Three independent experiments were carried out, with and without the addition of liver S9 mix from phenobarbital- and β- naphthoflavone induced rats (exogenous metabolic activation). In initial range-finding cytotoxicity test(s), the experimental doses of the main experiments were determined. Cells were exposed for 4 or 24 h to the test substance at concentrations from 0 to 1500 µg/mL. Vehicle and positive controls (ethyl methanesulphonate and 7,12 -dimethylbenzanthracene) were included as well in the experiments. The vehicle control showed mutation frequencies within the range expected for the CHO cell line. Both positive control substances led to the expected increase in the frequencies of forward mutations. In the 1st experiment in the absence and presence of metabolic activation and in the 2nd experiment in the presence of metabolic activation, the highest concentrations tested for gene mutations were clearly cytotoxic. However, in the 2nd experiment in the absence of metabolic activation no cytotoxicity was observed up to the highest applied concentration. Therefore, a 3rd experiment was performed which clearly showed reduced colony numbers at least at the highest concentrations. The test substance did not cause any relevant increase in the mutation frequency either without S9 mix or after the addition of a metabolizing system in the experiments. Under the study conditions, the substance was not mutagenic at the HPRT locus of Chinese hamster ovary cells (BASF SE, 2012).

Justification for classification or non-classification

Based on in vitro genetic toxicity studies with the substance itself and with the read-across substances L-glutamic acid, N-coco acyl derivs., monosodium salts and L-glutamic acid, N-coco acyl derivs., disodium salts, no classification for genetic toxicity is warranted according to EU CLP (1272/2008) criteria.