Registration Dossier

Administrative data

Key value for chemical safety assessment

Additional information

Only test data for the Ames test were available for registered substance, however read across data were available from read across substance Sodium bis(C11-14-isoalkyl, C13-rich) sulfosuccinate (CAS No. 848588 -96 -5). Justification for read across within the category of sulfosuccinates Di-esters is documented in a separate document attached in Section 13.

 

Bacterial reverse mutation

- In a key Ames test, the registered test item containing 63-67% active ingredient was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537, TA 1538, TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA (Müller, 1988). The mutagenicity studies were conducted in the absence and in the presence of a metabolizing system derived from rat liver homogenate. A dose range of 6 different doses from 4 to 5000µg/plate was used. Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in the literature. All the positive control compounds gave the expected increase in the number of revertant colonies. The test compound proved not to be toxic to the bacterial strains. 5000µg/plate was chosen as top dose level for the mutagenicity study. In the absence of the metabolic activation system the test compound did not show a dose dependent increase in the number of revertants in any of the bacterial strains. Also in the presence of a metabolic activation system, treatment of the cells with Netzer SB 10 did not result in relevant increases in the number of revertant colonies. It can be stated that the test substance is not mutagenic in these bacterial test systems either with or without exogenous metabolic activation at the dose levels investigated.

- In a supporting Ames test with -read across substance 'Sodium bis(C11-14-isoalkyl, C13-rich) sulfosuccinate' (CAS No. 848588 -96 -5), 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 (Flügge, 2013a). 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, which was also 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).

In conclusion, negative results were obtained for bacterial mutagenicity in a key study with registered substance and a support study with read across substance CAS No. 848588 -96 -5 tested in 5 strains with and without metabolic activation.

 

Mammalian mutagenicity

Data were available from read across substance 'Sodium bis(C11-14-isoalkyl, C13-rich) sulfosuccinate' (CAS No. 848588 -96 -5).

In a key mammalian gene mutation study (Flügge, 2013b) the test item was tested for mutagenic potential in a gene mutation assay in cultured mammalian cells (V79, genetic marker HPRT) both in the presence and absence of metabolic activation. The duration of the exposure with the test item was 4 hours or 24 hours in the experiments without S9 mix and 4 hours in the experiments with S9 mix.

The test item was completely dissolved in aqua ad iniectabilia. Aqua ad iniectabilia served as the vehicle control. Based on cytotoxicity at concentrations of 250 or 1000 µg/mL in the preliminary test, 500 µg test item/mL was employed as the top concentration for the main study.

Concentrations of 15.63, 31.3, 62.5, 125, 250 or 500 µg test item/mL and 31.3, 62.5, 125, 250 or 500 µg test item/mL were selected for the experiments without and with metabolic activation, respectively. The mutation frequency of the cultures treated were within the normal range of the vehicle controls. Under the present test conditions, the test item tested up to cytotoxic concentrations in the experiments without and with metabolic activation was negative in the HPRT-V79 mammalian cell mutagenicity test conditions where positive controls exerted potent mutagenic effects.

In conclusion,negative results were obtained for mammalian mutagenicity in a key study with read across substance CAS No. 848588 -96 -5

tested in V79 cells with genetic marker HPRT.

 

Chromosome aberration

Data were available from read across substance 'Sodium bis(C11-14-isoalkyl, C13-rich) sulfosuccinate' (CAS No. 848588 -96 -5).

A key in vitro micronucleus test was conducted using human peripheral lymphocytes both in the presence and absence of metabolic activation by a rat liver post-mitochondrial fraction (S9 mix) from Aroclor 1254 induced animals (Flügge, 2013c). The test was carried out employing 2 exposure times without S9 mix: (4 hours and 20 hours) and one exposure time with S9 mix (repeated). The harvesting time was 24 hours after the end of exposure. Each treatment was conducted in duplicate. The test item was completely dissolved inaqua ad iniectabilia. The vehicle aqua ad iniectabilia served as the vehicle control. Based on a preliminary experiment, cytotoxicity was noted starting at a concentration of 250 µg test item/mL in the experiment without and with metabolic activation. Hence, 250µg/mL were employed as the top concentration for the main test without and with metabolic activation in two independent experiments, each (4-hour and 20-hour exposure). There was no increase in micronuclei up to the cytotoxic concentration when compared to control both with and without metabolic activation. Under the present test conditions, the test item tested up to cytotoxic concentrations, in the absence and in the presence of metabolic activation employing two exposure times (without S9) and one exposure time (with S9) revealed no indications of any chromosomal damage in thein vitromicronucleus test. In the same test, Mitomycin C and cyclophosphamide induced significant damage.

In conclusion, negative results were obtained for chromosome aberration in a key study withread across substance CAS No. 848588 -96 -5 t ested in an in vitro Micronucleus test in human peripheral lymphocytes.

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, and no further testing is needed. The substance can be considered to have no mutagenic or genotoxic potential.


Justification for selection of genetic toxicity endpoint
Although the Ames study was selected as key study, the other studies were equivalent (key) studies for mammalian gene mutation and chromosome aberration.

Short description of key information:
Data were available from the registered substance and the read across substance Sodium bis(C11-14-isoalkyl, C13-rich) sulfosuccinate (CAS No. 848588 -96 -5). In the key Ames test with the registered substance, no increase in mutations were observed in different Salmonella typhimuriumstrains with and without metabolic activation up to 5000 µg/plate. In a supporting Ames test, the read across test item tested up to a concentration of 5000 µg/plate also 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 Mammalian gene mutation test the read across substance tested without and with metabolic activation was negative in the HPRT-V79 mammalian cell mutagenicity test conditions up to cytotoxic doses of 500 µg/mL. Finally In a key in vitro Micronucleus study in human peripheral lymphocytes, no indications of chromosomal damage were seen up to a cytotoxic concentration of 156.3 µg/mL medium, in the absence and in the presence of metabolic activation employing two exposure times (without S9) and one exposure time (with S9), respectively.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on the negative findings with registered and read across substance, the test item does not need to be classified and has no obligatory labelling requirement for genotoxicity according to the EC Directive (No.93/21/EEC) and CLP (No. 1272/2008 of 16 December 2008).