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Administrative data

Key value for chemical safety assessment

Additional information

In vitro data:

Mutagenicity in bacterial reverse mutation assays (Ames test) has been investigated with three members of the category of derivatives of 4,4’-bis(1,3,5-triazinyl-2-yl)amino)stilbene-2,2’-disulfonic acid, each with one anilino and one alkyl amino moiety: EC 432-690-8, EC 476-900-6 and EC 416-640-2. Negative results were obtained in all tests with and without metabolic activation.

Clastogenicity in mammalian cells has been investigated in reliable studies with three members of the category ofderivatives of 4,4’-bis(1,3,5-triazinyl-2-yl)amino)stilbene-2,2’-disulfonic acid, each with one anilino and one alkyl amino moiety: EC 432-690-8, EC 476-900-6 and EC 416-640-2. Negative results were obtained in the presence and absence of metabolic activation.

 

In vivo data:

A micronucleus test was investigated with the analogue substance EC 476-900-6. The oral application of the substance to mice did not induce micronuclei as determined by the micronucleus test in the bone marrow cells (polychromatic erythrocytes).

 

All these tests consistently yielded negative results indicating that the members of the category have no mutagenic potential. It is concluded that these negative findings can also be transferred to the substance defined in section 1. Besides these negative findings on mutagenicity the substance is unlikely to be bioavailable after oral, dermal and inhalation exposure and therefore has not to be classified as germ cell mutagens. No carcinogenic properties were identified.

 

 

EC 432-690-8:

None of the in vitro mutagenicity studies in bacteria and mammalian cells afforded indications of mutagenic effects neither with nor without metabolic activation. Neither gene mutations nor chromosomal aberrations were detected.

 

EC 476-900-6:
In vitro testing: The mutagenic effects of the test item were determined in a reverse mutation assay with Salmonella typhimurium. Test systems were the strains TA 97a, TA 98, TA 100., TA 102, and TA 1535 with (+) and without (-) the metabolic activation system S9 (from male Wistar rats). Concentrations tested were 0.003 - 0.01 - 0.1- 0.333 - 1.0 - 2.5 – 5.0 mg/plate. Three replicates per concentration level and control were performed. Based on the results of this study the test item was found to have no mutagenic effects.

The potential of the test item to induce chromosome aberrations was investigated in V79 Chinese hamster lung fibroblasts (V79) in vitro. For each experiment duplicate cell cultures were used for each concentration. The test compound was tested at concentrations of 46.9 to 2850 µg/mL including solvent controls with (+) and without (-) metabolic activation system from Spraque Dawley-rat liver. The test substance registered was clastogenic in V79 cells without S9 mix at the 28 hrs preparation interval, whereas in all other parts of the experiment no clastogenicity was observed. The observed increase of the aberration rate under this experimental conditions (28 hrs preparation interval without S9 mix) appeared together with an increase of cytotoxicity at these dose levels. It is striking, that the aberration rate increased with increasing cytotoxicity, indicating that clastogenicity correlated with cytotoxicity. As it has been shown for many chemicals that positive results are often irrelevant when correlated with cytotoxcity and that such chemicals are not genotoxic in vivo, the relevance of the present result with the test substance registered for the in vivo situation is questionable. Furthermore, in a vivo mutagenicity tests (i.e. the in vivo micronucleus test) it is proofed that the test substance is not genotoxic in vivo. Thus, the result of the chromosomal aberration study can be regarded as an irrelevant positive finding .

 

In vivo testing:The potential to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse was assessed in the micronucleus assay (OECD 474). The test substance was administered once orally to 12 mice per test group (6 per sex) at dose levels of 500, 1000 and 2000 mg/kg bw for the 24 hour preparation interval. A dose level of 2000 mg/kg bw was investigated for the 48 hour interval. The mean number of polychromatic erythrocytes was not decreased after treatment with the test item which indicates that the test substance did not have any cytotoxic properties in the bone marrow. In comparison to the respective controls there was no statistically significant or biologically relevant enhancement in the frequency of the detected micronuclei at any preparation interval and dose level after administration of the test item. The mean values of the micronuclei detected were below or near the value of the vehicle control. In parallel the treatment with the positive control (cyclophospahamide) showed a statistically significant increase of induced micronucleus frequency. Based on the experimental conditions reported the oral application of the substance to mice did not induce micronuclei as determined by the micronucleus test in the bone marrow cells (polychromatic erythrocytes).

The analogue substance EC 476-900-6 is not considered a genotoxin of relevance for humans.

 

EC 416-640-2

None of the in vitro mutagenicity studies in bacteria and mammalian cells afforded indications of mutagenic effects neither with nor without metabolic activation. Neither gene mutations nor chromosomal aberrations were detected.


Short description of key information:
The substance defined in section 1 has with high probability no mutagenic potential.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available studies are considered reliable and suitable for classification purposes under 67/548/EEC. As a result the category members are not considered to be classified for genetic toxicity under Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC.

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the category members are not considered to be classified for genetic toxicity under Regulation (EC) No. 1272/2008.