Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In a key Ames test the test item tested up to a concentration of 5000 µg/plate, caused no mutagenic effect in the Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 neither in the plate incorporation test nor in the preincubation test each carried out without and with metabolic activation. In a key Chromosome aberration toxicity study in V79 cell of the Chinese hamster in vitro, no structural chromosome and numerical aberrations were observed with and without metabolic activation mix when tested up to cytotoxic concentrations. In a key Mouse Lymphoma assay in the cell line L5178Y, the test item did not induce mutations in the mouse lymphoma thymidine kinase locus assay in the absence and presence of metabolic activation when tested up to cytotoxic concentrations.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Bacterial mutagenicity

In a key Ames test (Flügge, 2013) the test item was examined in the 5 Salmonella typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 in two independent experiments, each carried out without and with metabolic activation. The first experiment was carried out as a plate incorporation test and the second as a preincubation test. The test item was completely dissolved in aqua ad iniectabilia. Aqua ad iniectabilia was used as vehicle control. Based on a preliminary cytotoxicity test, six concentrations of 31.6, 100, 316, 1000, 3160 and 5000 µg test item/plate were employed in the plate incorporation test and in the preincubation test, each carried out without and with metabolic activation. No signs of cytotoxicity were noted in the plate incorporation test and in the preincubation test. No increase in revertant colony numbers as compared with control counts was observed for the test item, tested up to a concentration of 5000 µg/plate, in any of the 5 test strains in two independent experiments without and with metabolic activation, respectively (plate incorporation and preincubation test).

Mammalian gene mutation

In a key Mouse Lymphoma assay in the cell line L5178Y(Wollny, 2006), a test item containing 80% active ingredient was assessed for its potential to induce mutations in the mouse lymphoma thymidine kinase locus using the cell line L5178Y with and without liver microsomal activation and a treatment period of 4h. After a dose range finding study, two independent experiments with in duplo cell cultures were tested up to highly toxic concentrations leading to 10 -20% viability. No substantial and reproducible dose dependent increase in mutant colony numbers was observed in both main experiments. No relevant shift of the ratio of small versus large colonies was limited by toxicity of the test item. Under the experimental conditions reported, the test substance did not induce mutations in the mouse lymphoma thymidine kinase locus assay using the cell line L5178Y in the absence and presence of metabolic activation.

Chromosome aberration

In a key Chromosome aberration toxicity study in V79 cell of the Chinese hamster in vitro (Schulz, 2003), a test item containing 80% active ingredient was assessed for its potential to induce structural chromosome aberrations in two independent experiments up to 5000 µg act.ingr./mL (approx. 10 mM of the active ingredient). In each experimental group two parallel cultures were set up. Per culture 100 metaphase plates were scored for structural chromosome aberrations. Toxic effects indicated by reduced cell numbers and/or mitotic indices of about and below 50% of control were observed in all experimental parts. However, in experiment I in the absence and the presence of S9 mix concentrations showing clear cytotoxicity were not evaluable for cytogenetic damage. In both independent experiments, no biologically relevant increase in the number of cells carrying structural and numerical chromosomal aberrations was observed after treatment with the test item. Appropriate mutagens were used as positive controls.

Conclusion

Standard information requirements according to REACH Guidance Part 3 R7a were fulfilled for genotoxicity testing, including bacterial and mammalian mutagenicity and chromosomal aberration. Based on the available results, there were no indications of mutagenicity or genotoxicity. The substance can be considered to have no mutagenic or genotoxic potential.

An in vivo cytogenetics assay was requested based on ECHA Communication number CCH-D-2114330559-45-01/F. The results will be provided as soon as possible with an update of the dossier.Docusate sodium was used as read across substance for the registration of potassium 1,2-bis(2-ethylhexyloxycarbonyl)ethanesulphonate (CAS 7491-09-0). Based on the results of the in vitro chromosomal abberation (OECD 473 -) study wiht docusate sodium, which showed increases in the proportion of cells with structural aberrations, ECHA requested an in vivo cytogenetics assay (mammalian erythrocyte micronucleus test, mammalian bone marrow chromosomal aberration test or mammalian alkaline comet assay). The new study with docusate sodium will be conducted for both the registrations of bothpotassium 1,2-bis(2-ethylhexyloxycarbonyl)ethanesulphonate and docusate sodium, but also for the other diester sulfosuccinates.


Justification for classification or non-classification

Based on these results and according to the EC Directive (No.93/21/EEC) and CLP (No. 1272/2008 of 16 December 2008), the test substance does not have to be classified and has no obligatory labelling requirement for genetic toxicity.