Sodium 4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]toluene-3-sulphonate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Acid Violet 43 was tested in two different Amestests according to OECD guideline 471,both in strains TA 1535, TA 1537, TA 98 and TA 100 with and without metabolic activation.Materials of different purity (54.4% resp.95%) were used in these studies.In both experiments,the test substances did not increase the number of revertants in any strain in the presence or absence of S9.

In a mouse lymphoma assay according to OECD guideline 476,the test substance did not induce gene mutations in two independent experiments in cell line L5178Y at the tk^+/--locus with and without metabolic activation.In experiment 1 strong cytotoxicity was observed at the highest dose level tested after 4-hour treatment (±S9), and in experiment 2 relevant toxic effects were observed at the highest concentration tested under precipitation after 24-hour treatment (-S9).

Acid Violet 43 was tested in two different experiments in an in vitro chromosome aberration test in human lymphocytes according to OECD guideline 473.The highest concentration selected for each of these tests was the lowest concentration achieving a reduction of the mitotic index in the range 50-75%.For the 24-h harvest time the test substance did not induce any significant increase in the aberrant cells frequency, with and without metabolic activation in both experiments. For the 48-h harvest time performed during the repeat test,a significant increase in the aberrant cells frequency was recorded in the 2-h treatment group at the highest concentration with metabolic activation as well as in the 48-h treatment group at the highest concentration without metabolic activation. Aberrations consisted almost of chromatid deletions,and the proportion of cells bearing numerical aberrations was also slightly increased. These changes were observed together with a reduction in mitotic index to 46 and 52 % of controls.

According to the results of this study Acid Violet 43has the potential to induce chromosome aberrations in cultured mammalian cells.

In another chromosome aberration study according to OECD guideline 473,Acid Violet 43 was tested in two independent tests in Chinese Hamster Ovary Cells.The highest concentration for each experimental condition was selected on the basis of solubility or cytotoxicity criteria.In both experiments treatment of cultures with the test substance in the presence and absence of S9 and at both harvest times,resulted

in statistically significantly increased numbers of cells bearing structural aberrations, chromatid and/or chromosome deletions and exchanges. Some of these increases were observed in the absence of overt cytotoxicity.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)

Genetic toxicity in vivo

Description of key information

Acid Violet 43 was tested in an in vivo micronucleus study according to OECD guideline 474 on Swiss mice which received a single oral dose of 2000 mg/kg.In all groups treated with the test substance, the mean values of micronucleated polychromatic erythrocytes were similar to those of their respective vehicle group at each sampling time and no statistically significant differences were observed. At the 48-h sampling time, the PCE/NCE ratio was lower than in controls but this difference was mainly due to a high control value and does not clearly indicate bone marrow toxicity of the test substance.

 

To rule out any clastogenic potential of Acid Violet 43, the in vivo clastogenic potential was evaluated in an additional oral bone marrow micronucleus test in mice using conditions selected to maximize exposure of the animals.Acid Violet 43 was given to mice by gavage at the test limit dose level of 2000 mg active dye. There were no increases in the incidence of micronucleated polychromatic erythrocytes.There was no evidence of bone marrow toxicity but toxicokinetic evaluation confirmed systemic exposure of the animals and thus target organ exposure,since it is generally recognized that detection of test-related material in blood or plasma is adequate for validating bone marrow exposure .Accordingly, this additional in vivo bone marrow micronucleus test confirmed that Acid Violet 43 has no clastogenic potential in vivo.

Acid Violet 43 was also tested in an UDS-test according to OECD guideline 482 on

Wistar rats at dose levels of 82 and 816 mg active dye.Dose levels were selected on the basis of a sighting test in which 816 mg active active dye/kg was the Maximal Tolerated Dose, associated with piloerection and blue-coloured urine.

For each animal, hepatocytes were isolated from the liver and at least three primary cultures were established. Autoradiographic slides from 2 cultures/animal were prepared and the unscheduled synthesis of DNA was evaluated by the incorporation of tritiated methyl thymidine in 50 cells/slide.

No changes from controls in the number of nuclear and net grain counts were observed in Acid Violet 43 treated rats at both dose levels and both killing times of 2h and 16h respectively.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)