Reaction products of 2-amino-4,6-dinitrophenol, 4-amino-5-hydroxy 2,7-naphthalenedisulfonic acid, 4-nitrobenzenamine and resorcinol, sodium salts

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro Gene Mutation study in Bacteria - AMES

Positive, with and without metabolic activation. Under the test condition, the substance did induce point mutation by base pair change or frameshifts in the genome of the strains TA 1537, TA 1538, TA 98 and TA 100.

In vitro mammalian cells Chromosome aberration, OECD473

Negative. The substance, under the experimental conditions, did not induce structural chromosome aberrations in the V79 Chinese hamster cell line.

In vitro mammalian cell gene mutation assay, OECD476

Negative. Under the experimental condition, the substance did not induce point mutation at the HGPRT locus in V79 cells.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In vivo Unscheduled DNA Synthesis in rat hepatocytes

Negative. The substance did not induce DNA - damage leading to repair synthesis in the hepatocytes of the treated rats.

In vivo Mammalian Erythrocytes Micronucleous, OECD474

Negative. The test has been performed on mouse and rats. The test article did not induce micronuclei in mouse and rat.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Studies on bacteria (AMES)

The test has been conducted on the Similar substance 01, according to the OECD Guideline 471 and to the EU Method B.14 in order to evaluate the potential of the test item to induce gene mutations according to the plate incorporation test.

During the test, Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 have been used.

Under the test condition, the substance did induce point mutation by base pair change or frameshifts in the genome of the strains TA 1537, TA 1538, TA 98 and TA 100. The Similar substance 01 is considered to be mutagenic in this assay.

Based on the read-across principle(read-across from supporting substance -structural analogue or surrogate), the result can be considered for the genetic toxicity assessment of the registered substance. Justification for read-across is detailed in the report attached to the IUCLID section 13.

Studies on in vitro mammalian cells

Two tests have been conducted on Similar substance 01, according to the Guideline OECD 473

During the tests the V79 cell lines has been tested in presence and absence of the Mammalian microsomal fraction S9 mix (obtained from S9 liver microsomal fraction from Wistar rats)

The test item, under the experimental conditions, did not induce structural chromosome aberrations in the V79 Chinese hamster cell line, therefore is considered to be not mutagenic.

One test has been conducted on Similar substance 01, according to the OECD Guideline 476.

Under the experimental condition, the substance did not induce point mutation at the HGPRT locus in V79 cells, in presence and absence of the Mammalian microsomal fraction S9 mix. Therefore the Similar substance 01 is considered to be not mutagenic.

Based on the read-across principle (read-across from supporting substance-structural analogue or surrogate), the result can be considered for the genetic toxicity assessment of the registered substance. Justification for read-across is detailed in the report attached to the IUCLID section 13.

Studies in vivo

Two tests have been conducted according to the OECD Guideline 474 in mouse and rats. During the study the Similar substance 01 was assessed in the micronucleus assay for its potential to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse.

The mean values of micronuclei observed after treatment with the test article were in the same range as compared to the negative control groups in both species and did not induce micronuclei as determined by the micronucleus test in the bone marrow cells of the mouse.

Therefore, the Similar substance 01 is considered to be non-mutagenic in this micronucleus in vivo assay.

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

According to ECHA Guidance R.7a, Figure R.7.7-1 Flow chart of the mutagenicity testing strategy and Table R.7.7-5, a positive Gene mutation test in bacteria (AMES) requires additional evaluation on the potential mutagenicity of the substance, proceeding with Annex VIII (or Annex IX if in vivo testing is considered appropriate).

Studies on in vitro mammalian cells, performed according to OECD473 and OECD476 (Micronucleus and Chromosome aberration), are negative.

In addition, an in vivo a study performed according the OECD474 (Mammalian erythrocyte micronucleus) and an USD study (Unscheduled DNA synthesis test with mammalian live) are available, with negative results.

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 induced gene mutations in the strains in the Salmonella typhimurium reverse mutation assay.

However, additional studies are available, in vitro (Micronucleus and Chromosome aberration) and in vivo (Mammalian erythrocyte micronucleus test and Unscheduled DNA synthesis (UDS) test with mammalian liver). The studies are all negative.

Based on the read-across principle, the results can be 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.