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EC number: 201-549-0 | CAS number: 84-65-1
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
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- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
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- Toxicological Summary
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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:
- 9.6-9.8.2009
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: This study was carried out in accordance with internationally valid GLP principles.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 009
- Report date:
- 2009
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- yes
- Remarks:
- The deviations had no influence on study results.
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Anthraquinone
- EC Number:
- 201-549-0
- EC Name:
- Anthraquinone
- Cas Number:
- 84-65-1
- Molecular formula:
- C14H8O2
- IUPAC Name:
- anthraquinone
- Details on test material:
- Batch No.: V 1161
Purity: main component: 9,10-Anthraquinone 98.9 wt.%
Significant impurities: Anthracene 0.01 wt.%, 2,3-Naphthalic anhydride 0.25%, Naphtho[2,3-b]thiphenquinone 0.33%, 1,4- Anthraquinone 0.32%
Appearance: Yellow crystalline powder
Expiration date: 06/2024
Storage: The test substance was stored in the dark place at laboratory temperature during the study.
Constituent 1
Method
- Target gene:
- gene for synthesis histidine or tryptophan
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: histidine dependent strain
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- other: tryptophan dependent strain
- Metabolic activation:
- with and without
- Metabolic activation system:
- post-mitochondrial fraction (S9)
- Test concentrations with justification for top dose:
- 1.5, 5, 15, 50, 150, 1000 μg/plate
- Vehicle / solvent:
- - Solvent(s) used: DMSO
Controlsopen allclose all
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-o-phenylenediamine
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 9-aminoacridine hydrochloride monohydrate
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminofluorene
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: N-methyl-N´-nitro-N-nitrosoguanidine
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
NUMBER OF REPLICATIONS: Two series of experiments were performed with each strain - without metabolic activation and with a supernatant of rat liver and a mixture of cofactors.
DETERMINATION OF CYTOTOXICITY:
- Method: total growth - Evaluation criteria:
- The main criterion for evaluation of results was modified two-fold increase rule, its using is comparable with using of statistical methods (2, 3). After this rule the result is positive, when reproducible dose-effect and/or doubling of ratio Rt/Rc is reached.
- Statistics:
- Statistical method:
2. Dunkel V. C., Chu K.C. (1980): Evaluation of methods for analysis of microbial mutagenicity assays, in The Predictive Value of Short-Term Screening Tests in Carcinogenicity Evaluation, Elsevier North-Holland Biomedical Press, 231 - 240.
3. Claxton L. D. et al. (1987): Guide for the Salmonella typhimurium/mammalian microsome tests for bacterial mutagenicity, Mutat. Res. 189, 83 - 91.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- COMPARISON WITH HISTORICAL CONTROL DATA:
Spontaneous reversions, negative controls (solvent), and positive controls were compared with historical controls in our laboratory.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
Selection of doses/toxicity The test substance was dissolved in DMSO at 45ºC (temperature at which test substance is held in top agar before pouring onto Petri dishes) till the final concentration 150 µg/01 mL. This basic concentration was then diluted to gain a concentration range 1-150 µg/0.1 mL (plate). Because of the highest concentration was more than by digit position lower than the highest dose recommended by the guidelines, two other doses - 750 and 1500 µg per plate - were further prepared as suspensions of test substance in DMSO. Final concentration range 1-1500 µg per plate was then tested for toxicity in TA 100.
The test substance precipitated in top agar in doses 50 and 150 µg per plate. No signs of toxicity were observed. The dose 150 µg/0.1 mL (plate) stayed as maximum for dilution to concentration range 1-150 µg/0.1 mL (plate) in the first mutagenicity experiments. The highest dose of 1000 µg/plate was chosen as a dose between two other doses following according to rules (500 and 1500 µg/plate). It was additional, it was applied as a suspension and it caused cloud in top agar.
The highest dose (1000 µg/plate) used in the first mutagenicity experiments caused minor difficulties at evaluation caused by occurrence of suspension in background so it was not used for the second experiments. Instead, the other lower dose set according to the rules for dilution was added to the concentration range. In the the second mutagenicity experiments, 150 µg/0.1 mL stayed basic (the highest) concentration from what the concentration range was diluted. The solution was prepared by stirring the test substance in DMSO at the temperature of 45ºC approximately for an hour following by heating to 60ºC for approx. 5 minutes.
In the all experiments, the chosen doses were dosed in volume 0.1 mL per plate. Fresh solutions of test substance were prepared before each experiment. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
STUDY RESULTS
The results of experiments are summarized in tables IN Final report. The tables contain the dose applied per plate inµg (doses were applied to plates in volume 0.1 mL), amount of S9 per plate in µL numbers of revertants in single plates, average number of revertants per plate x and its standard deviation sd and ratio of revertants at tested dose (Rt) to number of revertants at negative control (Rc, dimethylsulfoxide).
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative without metabolic activation
ambiguous with metabolic activation
Under the above-described experimental design, the test substance Anthraquinone was nonmutagenic for all the Salmonella typhimurium as well as Escherichia coli strains both in experiments without as well as with metabolic activation. - Executive summary:
Test substance Anthraquinone was assayed for the mutagenicity by the Bacterial Reverse Mutation Test. The test was performed according to EU method B.13/14 Mutagenicity – Reverse mutation test using bacteria , which is analogous to the OECD Test Guideline No. 471.
Four indicator Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 and one indicator Escherichia coli WP2 uvrA strain were used. The test substance was diluted in dimethylsulfoxide and assayed in doses of 1.5-1000µg which were applied to plates in volume of 0.1 mL.
Two series of experiments were performed with each strain - without metabolic activation and with a supernatant of rat liver and a mixture of cofactors.
In the arrangement given above, the test substance Anthraquinone was nonmutagenic for all the used bacterial strains with as well as without metabolic activation.
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