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The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Additional information

There are no experimental data available on the Leucophor 1111X, thus a read across approach with the structural analogous has been proposed.

All the analogous substances considered are Stilbene derivatives Fluorescent Whitening Agents salts. They display similar structural and physicochemical properties; all of them exhibit high degree of dissociation in water and very low octanol/water partition coefficients because to a higher affinity with water phase than the octanol one. The water solubility is due to the sulphonate groups in the molecules.

The differences occurring in the structure formulas (i.e. substituents) are expected to not significantly impact the toxicological characterisation.

Bacterial reverse mutation assay (e.g. Ames test)

The Salmonella typhimurium and Escherichia coli Reverse Mutation Assay was performed to investigate the potential of the analogous substance 01 to induce gene mutations in the plate incorporation test (experiment I) and the pre-incubation test (experiment ll). The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The plates incubated with the test item showed normal background growth up to 5000 µg/plate with and without metabolic activation in both independent experiments. No toxic effects, evident as a reduction in the number of revertants, occurred in the test groups with and without metabolic activation. No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test item at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance. Appropriate reference mutagens were used as positive controls and showed a distinct in- crease of induced revertant colonies (Sokolowski, 2005).

The test result agrees with the outcomes of the other available studies performed on the analogous substance 02 and on the analogous substance 03.

Mammalian cell gene mutation assay

The analogous substance 02 was assayed for the mutagenicity by the In Vitro Mammalian Cell Gene Mutation Test. V79 hamster fibroblast were used for testing. A preliminary cytotoxicity assay (3-hour treatment) was performed at first; no toxicity was observed in experiments with metabolic activation in any dose, high toxicity was observed in the test without metabolic activation in the highest concentration only (survival 1 %).

The main experiments were performed without as well as with of metabolic activation using the supernatant of rat liver and a mixture of cofactors. Results of first experiment were negative, so the modification of experiment was performed with extended treatment time of 24 hours. The test was performed without metabolic activation only. The second expreriment gives no evidence of the mutagenicity of test substance. In conclusion, the test substance resulted non-mutagenic for V79 cells without as well as with metabolic activation (Täublová, 2015).

Chromosome aberration

The mammalian chromosome aberration potential has been assayed by in vitro system in Chinese hamster lung fibroblasts (V79) and Chinese hamster Ovary (CHO). Furthermore, an in vivo micronucleus assay in bone marrow cells of the mouse is also available.

The analogous substance 02 was tested for its potential to cause chromosome aberrations in cultured Chinese hamster ovary cells with and without an exogenous S-9 activation mixture. The test article was tested for toxicity and for the chromosome aberration potential at dose levels up to 5000 µg/ml. A parallel toxicity test was also performed. When compared to the solvent control data, the test doses did not cause a significant increase in the number of cells with chromosome aberrations in the non-activated or activated system (Thilagar, 1988).

The analogous substance 01 was assessed for its potential to induce structural chromosome aberrations in V79 cells of the Chinese hamster in vitro in four independent experiments. Evaluation of cytogenetic damage induced in V79 cells in the absence and the presence of metabolic activation was performed in four independent experiments. In each experimental group two parallel cultures were set up.

In the study, in the absence and the presence of S9 mix, toxic effects were observed at least at the highest applied concentrations, except for Experiment IB in the presence of S9 mix. However, in several experimental parts concentrations showing clear cytotoxicity were not scorable for cytogenetic damage. In Experiment IA, in the presence of S9 mix, increased aberration rates were observed at test item concentrations below the border of solubility in culture medium. Test item concentrations showing clear test item precipitation in culture medium were not scorable for genotoxicity. In the confirmatory experiment, designated Experiment IB, no clastogenicity was observed up to the highest scored concentration.

In Experiment IIA, at the 28 hrs preparation interval, after continuous treatment in the absence of S9 mix, biologically relevant increased aberration rates were observed. This observation was confirmed in the additional experiment, designated Experiment IIB, showing dose-related biologically relevant and statistically significant increases in the aberration rates. Under the experimental conditions reported, the test item induced structural chromosome aberrations as determined by the chromosome aberration test in V79 cells (Chinese hamster cell line) in vitro; therefore, the substance resulted to be clastogenic in this chromosome aberration test without S9 mix (Schulz and Kunz, 2006).

An in vivo study was performed in the same substance (analogous substance 01) in order to assess the potential of the test item to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse. The test item was formulated in deionised water, which was also used as vehicle control. The volume administered orally was 10 ml/kg b.w.. 24 h and 48 h after a single administration of the test item the bone marrow cells were collected for micronuclei analysis. Ten animals per test group were evaluated for the occurrence of micronuclei. At least 2000 polychromatic erythrocytes (PCEs) per animal were scored for micronuclei. To describe a cytotoxic effect due to the treatment with the test item the ratio between polychromatic and normochromatic erythrocytes was determined in the same sample and reported as the number of PCEs per 2000 erythrocytes. The following dose levels of the test item were investigated: 24 h preparation interval 500, 1000, and 2000 mg/kg b.w.; 48 h preparation interval 2000 mg/kg b.w.. After treatment with the test item the number of PCEs was not substantially decreased as compared to the mean value of PCEs of the vehicle control thus indicating that the substance did not exert any cytotoxic effects in the bone marrow. In comparison to the corresponding vehicle controls there was no biologically relevant or statistically significant enhancement in the frequency of the detected micronuclei at any preparation interval after administration of the test item and with any dose level used. Under the experimental conditions reported, the test item did not induce micronuclei as determined by the micronucleus test with bone marrow cells of the mouse (Honarvar, 2006).

Despite analogous substance 01 resulted to be clastogenic in chromosome aberration test without S9 mix, the substance is not considered as genotoxic because in presence of metabolic activation the substance dis not cause significant aberrations; furthermore, study outcomes from in vivo test confirmed that the test item can be considered as non genotoxic.

DNA damage and/or repair

The analogous substance 01 was assessed in the In vivo alkaline single cell gel electrophoresis analysis for its potential to induce primary DNA damage in cells prepared from the liver and small intestine of treated Wistar HsdCpb: WU (SPF) rats. At the highest dose clinical signs of toxicity such as ruffled fur and reduced spontaneous activity were observed. In each experimental group including the controls, hepatocytes and small intestine cells from six animals were assessed for the occurrence of DNA damage. None of the tested dose levels revealed a biologically relevant or statistically significant increase in DNA damage in the hepatocytes or the cells from the small intestine in the treated animals as compared to the corresponding vehicle controls in the evaluated parameter (% Tail Intensity). In addition, all % Tail intensities of the test item treated animals were within the historical control data range for the hepatocytes and even below the historical control range for the small intestine. In conclusion, oral administration of 1000 and 2000 mg test item/ kg b.w. did not induce any DNA-damage in the in vivo Comet assay performed in liver cells and cells from the small intestine isolated 4 or 16 hours after administration of the test item (Honarvar, 2008).

Justification for selection of genetic toxicity endpoint
Evaluation of the endpoint has been performed with the integrated evaluation of the following studies: in vitro Ames tests, in vitro gene mutation on mammalian cells, both in vitro and in vivo chromosomal aberration and in vivo alkaline single cell gel electrophoresis analysis in rat liver and small intestine.
Detailed justification for read across approach is given in the endpoint summary discussion and in the read across justification report attached to the Section 13 of this dossier.

Short description of key information:
Non genotoxic

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

According to CLP regulation (EC1272/2008), for the purpose of the classification for germ cell mutagenicity, substances are allocated in one of two categories in consideration of the fact that they are:

- substances known to induce heritable mutations or to be regarded as if they induce heritable mutations in the germ cells of humans or substances known to induce heritable mutations in the germ cells of humans or

- substances which cause concern for humans owing to the possibility that they may induce heritable mutations in the germ cells of humans.

On the basis of the results of the available studies, the substance can be considered as not having mutagenic or genotoxic properties.

In conclusion, the substance does not meet the criteria to be classified for genetic toxicity, according to the CLP Regulation (EC 1272/2008).