1,2-dihydroxyanthraquinone

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

The Mutagenic Constituents of Rubia tinctorum

Author:

Yoko Kawasaki et al.

Year:

1992

Bibliographic source:

Chemical Pharmacy Bulletin 40 (6) 1504-1509 (1992)

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Bacteria strain: Salmonella typhimurium TA98 and TA100

Principles of method if other than guideline:

The assay was carried out according to the pre-incubation method described by Yahagi et al.: In the preincubation assay, the tester strains are exposed to the chemical for a short time (20 to 30min) in a small volume (0.5ml) of either buffer or S-9 mix, prior to plating
on glucose agar minimal medium (GM agar) supplemented with a trace amount of histidine. With few exceptions it is believed that this assay is more sensitive than the plate incorporation assay, because short-lived mutagenic metabolites may have a better chance reacting with the
tester strains in the small volume of preincubation mixture, and the effective concentration of S-9 mix in the preincubation volume is higher than that on the plate.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,2-dihydroxyanthraquinone

EC Number:

200-782-5

EC Name:

1,2-dihydroxyanthraquinone

Cas Number:

72-48-0

Molecular formula:

C14H8O4

IUPAC Name:

1,2-dihydroxy-9,10-dihydroanthracene-9,10-dione

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Authentic 1,2-dihydroxyanthraquinone purchased from Wako Co. (Osaka)

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 100

Metabolic activation:

with and without

Test concentrations with justification for top dose:

300µg/plate

Vehicle / solvent:

0.1mL Dimethyl sulfoxide (DMSO)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Details on test system and experimental conditions:

Salmonella tiphymurium TA 98 and TA100 were supplied by Dr. T. Noumi of National Institute of Hygienic Sciences
Polychlorinated biphenyl induced rat liver S9 mixture: provided by Kikkoman Co. (Noda in Chiba Prefecture
Test sample (300µg was dissolved in 0.1mL of DMSO.

Evaluation criteria:

The mutagenicity was ranked as +++, ++ and + for activities of 10 or more, 5 or more and 2 or more times the spontaneous mutations frequency, respectively.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

not specified

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

not specified

Any other information on results incl. tables

Results of mutagenicity test:

	Mutagenicity*
	TA 98		TA 100
	- S9	+S9	-S9	+S9
Alizarin (1,2-dihydroxyanthraquinone) (300µg)	-	-	-	-

*The mutagenicity was ranked as +++, ++ and + for activities of 10 or more, 5 or more and 2 or more times the spontaneous mutations frequency, respectively.

Applicant's summary and conclusion

Conclusions:

Alizarin (1,2-dihydroxyanthraquinone) did not show any mutagenicity effect with and without metabolic activation at 300µg

Executive summary:

In this study the mutagenicity of authentic samples of alizarin (1,2 -dihydroxyanthraquinone) were investigated.

The assay was carried out according to the pre-incubation method: the tester strains are treated with the test substance for a short time (20 to 30min) in a small volume (0.5ml) of either buffer or S-9 mix prior to plating on glucose agar minimal medium (GM agar) supplemented with a trace amount of histidine. In this study, two strains of Salmonella typhimurium TA98 and TA100 were exposed to 300µg of 1,2 -dihydroxyanthraquinone with or without S9 -mix (metabolic activation). The test substance was dissolved in 0.1 mL of DMSO (dimethyl sulfoxide).

The results of this test shows clearly that the test substance does not possess mutagenicity effect on the strains tested (TA98 and TA100) with and without metabolic activation at a dose of 300µg.