17,18-dihydrodinaphtho[1',2',3':3,4;3'',2'',1'':9,10]perylo[1,12-efg][1,4]dioxocin-5,10-dione

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 Jan 2016-19 February 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

17,18-dihydrodinaphtho[1',2',3':3,4;3'',2'',1'':9,10]perylo[1,12-efg][1,4]dioxocin-5,10-dione

EC Number:

229-186-3

EC Name:

17,18-dihydrodinaphtho[1',2',3':3,4;3'',2'',1'':9,10]perylo[1,12-efg][1,4]dioxocin-5,10-dione

Cas Number:

6424-76-6

Molecular formula:

C36H18O4

IUPAC Name:

17,18-dihydrodinaphtho[1',2',3':3,4;3'',2'',1'':9,10]perylo[1,12-efg][1,4]dioxocin-5,10-dione

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Test item: Vat Blue 16

Method

Target gene:

The test item Vat Blue 16 was examined for the ability to induce gene mutations in tester strains of Salmonella typhimurium and Escherichia coli, as measured
by reversion of auxotrophic strains to prototrophy.

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

S9 liver homogenate from induced rat ( rat mixed induction).

Test concentrations with justification for top dose:

In Main Assay I : 1250, 625, 313, 156 and 78.1 µg/plate.
In Main Assay II : 625, 313, 156, 78.1 and 39.1 µg/plate.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

Toxicity and Main Assay I were perfomed using the plate incorporation method.
Main Assay II was perfomed using pre-incubation method.

Evaluation criteria:

For the test item to be considered mutagenic, two-fold (or more) increases in mean revertant numbers must be observed at two consecutive dose levels or at
the highest practicable dose level only. In addition, there must be evidence of a dose-response relationship showing increasing numbers of mutant colonies
with increasing dose levels.

Statistics:

Doubling rate (Chu et al. 1981).
Regression line.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

The test item did not induce two-fold increases in the number of revertant colonies, at any dose level, in any tester strain, in the absence or presence of S9 metabolism.

Applicant's summary and conclusion

Conclusions:

It is concluded that the test item Vat Blue 16 does not induce reverse mutation in Salmonella typhimurium or Escherichia coli in the absence or presence of S9 metabolism, under the reported experimental conditions.

Executive summary:

The test item Vat Blue 16 was examined for the ability to induce gene mutations in tester strains of Salmonella typhimurium and Escherichia coli, as measured by reversion of auxotrophic strains to prototrophy. The five tester strains TA1535, TA1537, TA98, TA100 and WP2 uvrA were used. Experiments were performed both in the absence and presence of metabolic activation, using liver S9 fraction from rats pre-treated with Phenobarbital and 5,6-Benzoflavone. The test item was used as a solution in dimethylsulfoxide (DMSO).

Toxicity test : Based on results obtained in a preliminary solubility trial, the test item Vat Blue 16 was assayed in the toxicity test at a maximum concentration of 1250 µg/plate and at four lower concentrations spaced at approximately half-log intervals: 395, 125, 39.5 and 12.5 µg/plate. No toxicity nor increases in revertant colonies were observed with any tester strain, at any dose level, in the absence or presence of S9 metabolic activation. Precipitation of the test item was observed at the end of the incubation period, with all tester strains, at the highest dose level tested, both in the absence and presence of S9 metabolism.

Main Assay I : On the basis of toxicity test results, in Main Assay I, using the plate incorporation method, the test item was assayed at the following dose levels: 1250, 625, 313, 156 and 78.1 µg/plate in the absence or presence of S9 metabolism. No toxicity was observed with any tester strain at any dose level, in the absence or presence of S9 metabolism. Precipitation of the test item was observed at the end of the incubation period, with all tester strains, at the highest dose level tested, both in the absence and presence of S9 metabolism.

Main Assay II : As no relevant increase in revertant numbers was observed at any concentration tested, a pre-incubation step was included for all treatments of Main Assay II. As limited by the maximum feasible concentration in DMSO, the test item was assayed at the following dose levels: 625, 313, 156, 78.1 and 39.1 µg/plate. No toxicity was observed with any tester strain, at any dose level, in the absence or presence of S9 metabolism. No precipitation of the test item was observed at the end of the incubation period at any concentration.

Conclusion : It is concluded that the test item Vat Blue 16 does not induce reverse mutation in Salmonella typhimurium or Escherichia coli in the absence or presence of S9 metabolism, under the reported experimental conditions.