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EC number: 235-436-2 | CAS number: 12227-50-8
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- 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
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1984
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 984
- Report date:
- 1984
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- other: Ames, B. N.; McCann, J.; Yamasaki, E.: Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mut. Res. 31 (1975) 347-364.
- Deviations:
- no
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 4',4'''-azobis[N-(9,10-dihydro-9,10-dioxo-1-anthryl)[1,1'-biphenyl]-4-carboxamide]
- EC Number:
- 235-436-2
- EC Name:
- 4',4'''-azobis[N-(9,10-dihydro-9,10-dioxo-1-anthryl)[1,1'-biphenyl]-4-carboxamide]
- Cas Number:
- 12227-50-8
- Molecular formula:
- C54H32N4O6
- IUPAC Name:
- 4',4'''-azobis[N-(9,10-dihydro-9,10-dioxo-1-anthryl)[1,1'-biphenyl]-4-carboxamide]
- Test material form:
- solid: particulate/powder
- Details on test material:
- Vat Yellow 33
Constituent 1
Method
- Target gene:
- histidine
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Test Concentrations for preliminary toxicity test & mutation study:
0.8, 4, 20, 100, 500 μg/well
The test product was found to have a maximum solubility of approximately 5,000 μg/ml but at levels of 500 μg/well and below was found to be non-toxic to the tester strains.
Accordingly, dose levels ranging from 0.8 μg/plate to 500 μg/plate were selected for the mutation study. - Vehicle / solvent:
- For the purpose of this study the test material was dissolved/suspended in Dimethyl sulphoxide.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- other: M-Methyl-N-Nitro-N-Nitrosoguanidine (MNNG); 4 Nitro-O-phenyldiamine (4NOPD); 2-aminoanthracene (2AA)
- Details on test system and experimental conditions:
- Bacteria
The strains used in this assay were all mutants derived from Salmonella typhimurium LT2 and were those recommended for general screening.
TA 1535 - sensitive to agents inducing base-pair substitution
TA 100 - sensitive to agents inducing base-pair substitution
TA 1537 - sensitive to agents inducing frame-shift mutations
TA 98 - sensitive to agents inducing frame-shift mutations
The bacteria were obtained from Professor B Ames (Department of Biochemistry, University of California, Berkeley, California, U.S.A.). Overnight cultures of these specimens were used to generate Master slopes which were maintained at 4°C for use as routine sources of inoculum for cultures to be used in this assay.
Prior to being used in this test characterisation checks were carried out on each strain to determine (i) histidine requirement (ii) crystal violet sensitivity (iii) presence of R factor, and (iv) spontaneous reversion rate.
In this assay overnight sub-cultures of the Master slopes were prepared in nutrient broth (Oxoid Ltd) incubated at 37 °C for approximately 18 hours.
These overnight cultures yielded approximately 10E+8 – 10E+9 bacteria per ml and were used as the standard bacterial suspension.
Microsomal Enzyme Fraction
A commercial preparation of this enzyme fraction was obtained from Uniscience Limited, 8 Jesus Lane, Cambridge, CB5 8BA. This fraction is obtained from animals pre-treated with Arochlor 1254 (a mixture of polychlorinated biphenyls) and was stated to be from batch no. 03121 protein level 25 mg/ml.
Test Procedure
a) Preliminary Toxicity Study
In order to select appropriate dose levels for use in the main study, a preliminary test was carried out to determine the toxicity of the test compound to the tester organisms. 0.1 ml of bacterial suspension was added to 4 ml of histidine deficient media (histidine & top agar) and overlayed onto sterile plates of Vogel-Bonner agar (minimal agar - 15 ml/plate). When the agar had set, wells were made on each plate and 0.1 ml aliquots of concentrations ranging from 8 μg/ml to 5,000 μg/ml of test material were added to each well. These plates were incubated at 37 °C for 24 hours after which time the toxicity of the test material was assessed by measuring the zones of inhibition around each well.
Dimethyl Sulphoxide which was used as a solvent diluent for the test material in this assay was also added to each plate as a control.
Mutation Study
Four concentrations of the test material were assayed in triplicate against each tester strain, with and without metabolic activation.
0.1 ml of the appropriately diluted test material, negative or positive control solution were placed in sets of sterile universal bottles containing 9 ml of molten histidine deficient top agar at 45 °C, these sets comprising two bottles for each tester strain/test material or control solution. A 0. 1 ml aliquot of one of the bacterial suspensions was added to each of the bottles. Into one of the duplicate sets of bottles was placed 0 .5 ml of the S-9 liver microsone mix; in the other set the S-9 mix was omitted. The contents of the bottle were then mixed and a 3 ml aliquot of each poured on top of Vogel-Bonner agar plates.
These plates were then incubated at 37 °C for 48 hours and the number of revertant colonies counted. - Rationale for test conditions:
- In accordance with the test procedures.
- Evaluation criteria:
- For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate (of at least twice the spontaneous reversion rate) in one or more strains of bacteria in the presence and/or absence of the S9 microsomal enzymes. To be considered negative the number of induced revertants compared to spontaneous revertants should be less than two fold at all dose levels employed, the intervals of which should be between 3 and 5 fold and extend to the limits imposed by toxicity or solubility.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Preliminary toxicity study
The test product was found to have a maximum solubility of approximately 5,000 μg/ml but at levels of 500 μg/well and below was found to be non-toxic to the tester strains.
Accordingly, dose levels ranging from 0.8 μg/plate to 500 μg/plate were selected for the mutation study.
Mutation study
The overnight culture of each strain was found to be in the required range of 10E+8 to 10E+9 bacteria/ml and the spontaneous reversion rate for each was found to be within the expected range.
No significant increase in the number.of revertant colonies was recorded for any of the strains at any of the dose levels employed in this study, either with or without metabolic activation. All counts of revertant colonies were similar to those recorded for the negative control plates and were within the range expected for spontaneous reversion for each strain.
The positive control substances all produced significant increases in the number of revertant colonies and the activity of the S9 fraction was found to be satisfactory.
Applicant's summary and conclusion
- Conclusions:
- Vat Yellow 33 was found to exhibit no evidence of mutagenic activity under the conditions of this experiment
- Executive summary:
This study was conducted according to Safepharm Standard Test Method M26 and was designed to assess the mutagenic potential of the test material using a bacterial/microsome test system. The study was based on the in vitro technique described by Ames and his co-workers and Garner et al in which mutagenic activity is assessed by exposing histidine auxotrophs of Salmonella typhimurium to various concentrations of the test compound in the presence and absence of S9-mix from induced rat livers.. The strains used in this assay were all mutants derived from Salmonella typhimurium LT2 and were those recommended for general screening: TA 1535, TA 100, TA 1537, TA 98.
In order to select appropriate dose levels for use in the main study, a preliminary test was carried out to determine the toxicity of the test compound to the tester organisms. In the main study, four concentrations of the test material were assayed in triplicate against each tester strain, with and without metabolic activation.
In the preliminary toxicity study, the test substance was found to have a maximum solubility/suspendability of approximately 5,000 μg/ml. At levels of 500 μg/well and below, the test substance was found to be non-toxic to the tester strains. Accordingly, dose levels ranging from 0.8 μg/plate to 500 μg/plate were selected for the mutation study.
I the main study, the overnight culture of each strain was found to be in the required range of 10E+8 to 10E+9 bacteria/ml and the spontaneous reversion rate for each was found to be within the expected range. No significant increase in the number of revertant colonies was recorded for any of the strains at any of the dose levels employed in this study, either with or without metabolic activation. All counts of revertant colonies were similar to those recorded for the negative control plates and were within the range expected for spontaneous reversion for each strain. The positive control substances all produced significant increases in the number of revertant colonies and the activity of the S9 fraction was found to be satisfactory.
Consequently, the test substance was found to exhibit no evidence of mutagenic activity under the conditions of this experiment.
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