Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro Gene Mutation study in Bacteria - AMES

Negative. The substance is not mutagenic for bacteria

In vitro mammalian cells Chromosome aberration, OECD473

Negative. The substance is not clastogenic in human lymphocytes.

In vitro mammalian cell gene mutation assay, OECD476

Test substance does not induce gene mutation in Chinese hamster V79 cells after in vitro treatment in the absence or presence of S9 metabolic activation.

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

For substances covered by Annex VIII to REACH genetic toxicity on bacteria and two tests on in vitro mammalian cells are required.

The tests required to fulfil the ANNEX VIII were performed on a similar substance 01 and Similar substance 02 (read-across from supporting substance -structural analogue or surrogate).

Studies on bacteria (AMES)

The mutagenicity on bacteria was performed following the OECD 471 and the EU B.13/B.14.

The similar substance 01 was tested in the Salmonella typhimurium reverse mutation assay with tour histidine- requiring strains of Salmonella typhimurium (TA1535, TA1537, TA100 and TA98) and in the Escherichia coli reverse mutation assay with a tryptopham requiring strain of Escherichia coli WP2uvrA in two independent experiments, modified according to Prival and Mitchell.

The substance was tested up to concentrations of 5000 pg/plate in the absence and presence of S9 mix.

The test item did not precipitate on the plates at this dose level.

The bacterial background lawn was not reduced at all concentrations tested and no decrease in the number of revertants was observed.

The substance did not induce a dose-related increase in the number of revertant (His*) colonies in each of the tour tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trpt) colonies in tester strain WP2uvrA both in the absence and presence of S9- metabolic activation. These results were confirmed in an independently repeated experiment.

Based on the results of this study it is concluded that the similar substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Studies on in vitro mammalian cells

The test has been performed according to the OECD 473 and EU B10.

In the first cytogenetic assay, the substance was tested up to 5000 pg/mL for a 3 h exposure time with a 24 h fixation time in the absence and presence of S9-mix.

In the second cytogenetic assay, the substance was tested up to 1500 pg/mL for a 24 h and 48 h continuous exposure time with a 24 h and 48 h fixation time in the absence of S9-mix.

Appropriate toxicity was reached at these dose levels. In the presence of 1.8% (v/v) S9-fraction was tested up to 5000 pg/ml for a 3 h exposure time with a 48 h fixation time.

The Similar substance 01 did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations in the absence and in the presence of S9-mix, in two independently repeated experiments.

It is concluded that this test is valid and that the Similar substance 01 is not clastogenic in human lymphocytes under the experimental conditions described in this report.

The GENE MUTATION IN CHINESE HAMSTER V79 CELLS OECD476- RTC Study Number: A2931.

It is concluded that the test substance does not induce gene mutation in Chinese hamster V79 cells after in vitro treatment in the absence or presence of S9 metabolic activation, under the reported experimental conditions.

Based on the read-across principle, this evaluated conclusion can be considered valid for the genetic toxicity assessment of the substance. Justification for Read Across is detailed in the report attached to the IUCLID section 13.

Justification for classification or non-classification

This hazard class is primarily concerned with substances that may cause mutations in the germ cells of humans that can be transmitted to the progeny.

Substance that are mutagenic in somatic cells may produce heritable effects if they, or their active metabolites, have the ability to interact with the genetic material of germ cells. Conversely, substances that do not induce mutations in somatic cell in vivo would not be expected to be germ cell mutagens.

However, the results from mutagenicity or genotoxicity tests in vitro and in mammalian somatic and germ cells in vivo are also considered in classifying substances and mixtures within this hazard class.

Category 1: substances known to induce heritable mutations or to be regarded as if they induce heritable mutations in the germ cells of humans. Substances known to induce heritable mutations in the germ cells of humans.

Category 2: substances which cause concern for humans owing to the possibility that they may induce heritable mutations in the germ cells of humans.

Classification for heritable effects in human germ cells is made on the basis of well conducted, sufficiently validated tests as In vitro mutagenicity tests such as these indicated in 3.5.2.3.8:

- in vitro mammalian chromosome aberration test;

- in vitro mammalian cell gene mutation test;

- bacterial reverse mutation tests

The Similar substance 01 did not induced gene mutations in the strains in the Salmonella typhimurium reverse mutation assay.

However, additional study is available, in vitro, Chromosome aberration, resulted negative.

Based on the Annex VIII, the missing GENE MUTATION IN CHINESE HAMSTER V79 CELLS OECD476-RTC Study Number: A2931, the test substance does not induce gene mutation after in vitro treatment in the absence or presence of S9 metabolic activation, under the reported experimental conditions.

Based on the read-across principle, the available results are considered for the genetic toxicity assessment of the substance.

As conclusion, according to the CLP Regulation n.1272/2008 and the ECHA Guidance R.7a, the substance is not classified as mutagenic.