Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test

The test substance (AD-2000) was tested in the Salmonella typhimurium reverse mutation assay with four histidine­ requiring strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli (WP2uvrA). The test was performed in two independent experiments  in the presence and absence of S9-mix (rat liver S9-mix induced by a combination of phenobarbital  and ß-naphthoflavone).

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

AD-2000 did not induce a significant dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+) colonies in tester strain WP2 uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed in an independently  repeated experiment

Based on the results of the dose range finding test, AD-2000 was tested in the first mutation assay at a concentration  range of 3 to 333 µg/plate in the absence and presence of 5% (v/v)

S9-mix in tester strains TA1535, TA1537 and TA98. In an independent  repeat of the assay with

additional parameters,  AD-2000 was tested at the same concentration  range as the first assay in the absence and presence  of 10% (v/v) S9-mix in tester strains TA1535, TA1537, TA98, TA100 and WP2 uvrA. AD-2000 precipitated on the plates at the top dose of 333 µg/plate. The bacterial background  lawn was not reduced at any of the concentrations  tested and no biologically relevant decrease in the number of revertants was observed

Choromosomal aberration study

This report describes the effect of the test substance on the number of chromosome aberrations in cultured peripheral human lymphocytes in the presence and absence of a metabolic activation system (phenobarbital and ß-naphthoflavone induced rat liver S9-mix). The possible clastogenicity of the test substance was tested in two independent experiments.

In the first cytogenetic assay, the test substance was tested up to 33 µg/ml for a 3 h exposure time with a

24 h fixation time in the absence and presence of 1.8% (v/v) S9-fraction. the test substance precipitated in the culture medium at this dose level.

In the second cytogenetic assay, the test substance was tested up to 100 µg/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. In the presence of S9-mix the test substance was tested up to 33 µg/ml for a 3 h exposure time with a 48 h fixation time. the test substance precipitated in the culture medium at these dose levels

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

No effects of the test substance on the number of polyploid cells and cells with endoreduplicated chromosomes were observed both in the absence and presence of S9-mix. Therefore it can be concluded that the test substance does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions described in this report

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

Additional information

Justification for classification or non-classification