Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation (Bacterial reverse mutation assay / Ames test): negative with and without metabolic activation in Salmonella typhimurium strains TA1535, TA1537, TA100 and TA98 and Escherichia coli WP2 uvrA (OECD TG 471) (Verspeek-Rip, C., 1999).
Cytogenicity in mammalian cells: not required: in vivo micronucleus study available
Mutagenicity in mammalian cells: negative with and without metabolic activation in mouse lymphoma L5178Y cells (WIL, 2015).

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

Genetic toxicity in vivo

Description of key information

Mouse micronucleus assay study (ip administration): Negative (OECD TG 474) (BioReliance, 2009).

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

Additional information

Information is available for mutagenicity to bacterial and mammalian cells from in vitro studies, and for cytogenicity from an in vivo micronucleus assay.

D6 has been tested in a reliable study conducted in accordance with OECD 471 and in compliance with GLP (Verspeek-Rip, C., 1999). No evidence of test substance induced increase in the number of revertants was observed when the substance was tested in the presence and absence of metabolic activation in Salmonella typhimurium strains TA1535, TA1537, TA100 and TA98 and Escherichia coli WP2uvrA, in either the initial or the independent repeat experiment. Appropriate solvent and positive controls were included and gave expected responses. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

In vitro cytogenicity testing is not required as there is a reliable in vivo micronucleus assay available.

In vitro mutagenicity: D6 has been tested for mutagenicity in mouse lymphoma cells L5178Y cells in a reliable study conducted according to OECD TG 476, in compliance with GLP (WIL, 2015a). No cytotoxicity was observed, and no increase in mutant frequency above the Global Evaluation Frequency was observed in either the absence or presence of exogenous metabolic activation when tested up to concentrations limited by the solubility of the test substance. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity in mouse lymphoma L5178Y cells under the conditions of the test.

D6 has been tested in a reliable in vivo micronucleus assay in ICR mice conducted according to OECD TG 474 and in compliance with GLP (BioReliance, 2009). No statistically significant increase in the incidence of micronucleated polychromatic erythrocytes relative to control was observed in any test substance group up to limit concentrations at 24 and 48 hours after intraperitoneal administration of the test substance. Reductions in PCE/EC ratio (up to 11%) were observed in some test article groups, indicating that the test article had reached the target tissue. Administration of cyclophosphamide (the positive control) induced marked increases in micronucleated polychromatic erythrocytes. It is concluded that the test substance is negative for the induction of micronuclei in vivo under the conditions of the test.





Justification for classification or non-classification

Based on reliable in vitro and in vivo data, D6 does not require classification for mutagenicity according to Regulation (EC) No 1272/2008.