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Toxicological information

Genetic toxicity: in vitro

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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:
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

Reference
Name:
Unnamed
Type:
Constituent
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 / strainopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Test concentrations with justification for top dose:
300µg/plate
Vehicle:
0.1mL Dimethyl sulfoxide (DMSO)
Controls
Negative controls:
yes
Solvent controls:
not specified
True negative controls:
not specified
Positive controls:
not specified
Details on test system and 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 resultsopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
not specified
Negative controls valid:
yes
Positive controls valid:
not specified
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
not specified
Negative controls valid:
yes
Positive controls valid:
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.