1,4-diamino-2,3-diphenoxyanthraquinone

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

In an Ames test MACROLEX Rotviolett R was using Salmonella typhimurium LT2 mutants TA 1535, TA 100, TA 1537, TA 98 and TA 102.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,4-diamino-2,3-diphenoxyanthraquinone

EC Number:

229-066-0

EC Name:

1,4-diamino-2,3-diphenoxyanthraquinone

Cas Number:

6408-72-6

Molecular formula:

C26H18N2O4

IUPAC Name:

1,4-diamino-2,3-diphenoxyanthraquinone

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Details on mammalian cell type (if applicable):

Histidine deficient mutants - riginal strains were obtained from Prof. Bruce ames

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

0, 16, 50, 158, 500, 1581 or 5000 µg/plate
0, 16, 50, 158, 500, 1581 or 5000 µg/tube

Vehicle / solvent:

DMSO (dimethylsulphoxide)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

cumene hydroperoxide

other: 4-nitro-1,2-phenylene diamine, 2-aminoanthracene

Details on test system and experimental conditions:

MACROLEX Rotviolett R was initially screened with one plate per dose using the Salmonella/microsome plate incorporation test for point mutagenic effects in doses of up to and including 5000 µg per plate on five Salmonella typhimurium LT2
mutants. These comprised the histidine-auxotrophic strains TA 1535, TA 100, TA 1537, TA 98 and TA 102.
Additionally MACROLEX Rotviolett R was screened in an independent repeat using the Salmonella/microsome test for point mutagenic effects in doses up to 5000 µg per tube after preincubation for 20 minutes at 37°C. All other conditions remained unchanged.

Evaluation criteria:

A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA 1535, A 100 and TA 98 this increase should be about twice that of negative controls, whereas for A 1537, at
least a threefold increase should be reached. For TA 102 an increase of about 150 mutants should be reached. Otherwise, the result is evaluated as negative. However, these guidelines may be overruled by good scientific judgement.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA 98, TA 100, TA 102, TA 1535, TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: Plate incorporation test: Doses up to and including 500 µg per plate did not cause any bacteriotoxic effects. Preincubation test: Doses up to and including 1581µg per tube did not cause any bacteriotoxic effects.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Evidence of mutagenic activity of MACROLEX Rotviolett R was not seen. No biologically relevant increase in the mutant count, in comparison with the negative controls, was observed.

Applicant's summary and conclusion

Conclusions:

Negative

Executive summary:

In an Ames test MACROLEX Rotviolett R was using Salmonella typhimurium LT2 mutants TA 1535, TA 100, TA 1537, TA 98 and TA 102. MACROLEX Rotviolett R was negative (non-mutagenic) without and with S9 mix in the plate incorporation as well as in the preincubation modification of the Salmonella/microsome test.