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EC number: 274-410-5 | CAS number: 70210-13-8
- 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
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1993
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 993
- Report date:
- 1993
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Trisodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonatonaphthyl]azo]-2,5-dimethylphenyl]azo]benzene-1,4-disulphonate
- EC Number:
- 274-410-5
- EC Name:
- Trisodium 2-[[4-[[4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulphonatonaphthyl]azo]-2,5-dimethylphenyl]azo]benzene-1,4-disulphonate
- Cas Number:
- 70210-13-8
- Molecular formula:
- C27H22ClN9O9S3.3Na
- IUPAC Name:
- trisodium 2-{[4-({4-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-5-sulfonato-1-naphthyl}diazenyl)-2,5-dimethylphenyl]diazenyl}benzene-1,4-disulfonate
- Test material form:
- other: solid
Constituent 1
- Specific details on test material used for the study:
- Test material: Cibacron Orange P-4R flüssig (FAT 40075/C)
Batch No.: 289637.26
Purity: 21.4 % (active ingredient)
Expiration date: January 1994
Method
- Target gene:
- Histidine gene
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):
- The histidine-auxotrophic strains of Salmonella typhimurium (TA 98, TA 100, TA 102, TA 1535 and TA 1537) were obtained from Prof. B. Ames, Berkeley, CA., U.S.A.
Inoculates from frozen master copies were set up monthly. They were grown in liquid NB-medium overnight and then plated on NB-agar. After incubation, single colonies were taken from the plates, grown overnight in liquid NB-medium and then used for the experiment.
The characteristics of the strains were checked monthly. Histidine-auxotrophy of the strains was demonstrated by the requirement for 1-histidine. The presence of the rfa character was assayed by the sensitivity for crystal-violet. The deletion of the uvrB gene (strains TA 98, TA 100, TA 1535 and TA 1537) was demonstrated by the sensitivity for UV-light. The Salmonella strains containing the R-factor (TA 98 and TA 100) were additionally checked for ampicillin resistance. Strain TA 102 additionally was checked for tetracycline resistance (presence of multicopy plasmid pAQ1). The presence of the uvr+ gene was demonstrated by the resistance of strain TA 102 against UV light. Furthermore, all strains were checked for their characteristic reversion properties with known mutagens (positive controls). - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Cytotoxicity test: 20.6, 61.7, 185.2, 555.6, 1666.7 and 5000 µg active ingredient/plate
Mutagenicity tests: 61.7, 185.2, 555.55, 1666.7 and 5000 µg active ingredient/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Bidistilled water
- Justification for choice of solvent/vehicle: Based on solubility
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- Bidistilled water
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- for TA 100 and TA 1535 without metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- bidistilled water
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- for TA 102 without metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- DMSO
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- for TA 98 without metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- DMSO
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- for TA 1537 without metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- DMSO
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- for TA 100, TA 102, TA 98 and TA 1537 with metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- bidistilled water
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide H20
- Remarks:
- for TA 1535 with metabolic activation
- Details on test system and experimental conditions:
- Setting up of the test plates:
0.1 mL of the overnight cultures were mixed with 2 mL of top agar, either 0.5 mL of 100 mM sodium phosphate buffer (experiments without activation) or 0.5 mL of the activation mixture (experiments with activation) and 0.1 mL of a solution of the test substance, the substance for the positive control or the solvent for the negative control and poured on minimal agar in Petri dishes. Each Petri dish contained about 20 mL of minimal agar (1.5 % agar supplemented with 2 % salts of the Vogel-Bonner Medium E and 2 % glucose). The top agar was composed of 0.6 % agar and 0.6 % NaCl. It was supplemented with 10 % of 0.5 mM 1-histidine and 0.5 mM (+) biotin dissolved in water.
Preliminary Toxicity/Range-Finding test:
A toxicity test (check for reduction in the number of revertant colonies) was carried out with strain TA 100 without and with microsomal activation at six concentrations of the test substance and one negative control according to Standard Operating Procedures of Genetic Toxicology. The highest concentration applied was 5000 µg/plate. The five lower concentrations decreased by a factor of 3. The plates were inverted and incubated for about 48 hours at 37 ± 1.5 °C in darkness. Thereafter, they were evaluated by counting the colonies and determining the background lawn. One plate per test substance concentration, as well as each negative control was used.
Mutagenicity test:
The mutagenicity test was performed with strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 without and with microsomal activation according to Standard Operating Procedures of Genetic Toxicology. Each of the five concentrations of the test substance, a negative and a positive control were tested, using three plates per test substance concentration as well as each positive and negative control with each tester strain. The highest concentration applied in the first and second mutagenicity test was 5000 µg/plate (because of lack of toxicity in the range finding test) and the four lower concentrations were each decreased by a factor of 3. The plates were inverted and incubated for about 48 h at 37 ±1.5 °C in darkness. Thereafter, they were evaluated by counting the number of colonies and determining the background lawn. - Evaluation criteria:
- Assay acceptance criteria: A test is considered acceptable if the mean colony counts of the control values of all strains are within the acceptable ranges and if the results of the positive controls meet the criteria for a positive response.
- Statistics:
- In deviation to the OECD guideline, a statistical analysis was not performed. At present the use of statistical methods concerning this particular test system is not generally recommended. No appropriate statistical method is available.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535 pSK1002
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- marginal response
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- marginal response
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- 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
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- Toxicity test/Range finding test: Six concentrations of the test substance ranging from 20.6 to 5000 µg active ingredient/plate were tested with strain S. typhimurium TA 100 to determine the highest concentration to be used in the mutagenicity assay. The experiments were performed with and without microsomal activation. Normal back-ground growth was observed at all concentrations. A slight increase in the number of revertant counts was observed at the upper concentrations, followed by a reduction in the number of revertant colonies at the highest concentration, due to toxicity of the test substance. From the results obtained, the highest concentration suitable for the mutagenicity test was selected to be 5000 µg active ingredient/plate without and with activation.
Mutagenicity test, original experiment: In the original experiment carried out without metabolic activation, treatment of strain TA 102 with the test substance led to a slight increase in the number of revertant counts at the concentrations of 185.2 to 1666.7 µg active ingredient/plate. In the experiment with activation, a similar effect occurred on strain TA 100 at concentration of 1666.7 µg active ingredient/plate and on strain TA 102 at concentrations of 185.2 and 1666.7 µg active ingredient/plate. No effects were observed with the other strains.
Mutagenicity test, confirmatory experiment: In the confirmatory experiment carried out without metabolic activation, treatment of strain TA 102 with the test substance led to a slight increase in the number of revertant counts at the concentration of 1666.7 µg active ingredient/plate. In the experiment with activation, a similar effect occurred on strains TA 100 and TA 102 at the same concentration. Again, no effects were observed with the other strains. In the mutagenicity tests normal background growth was observed at all concentrations. In the experiments without and with microsomal activation, due to toxicity of the test substance the numbers of revertant colonies occasionally were slightly reduced with strains TA 100 and TA 102 at the upper concentrations.
Applicant's summary and conclusion
- Conclusions:
- Under the study conditions, the test substance is considered to be non-mutagenic in the Salmonella typhimurium reverse mutation assay.
- Executive summary:
An in vitro study was performed to investigate the potential of the test substance (of ca. 21.4 % purity) to induce gene mutations according to OECD Guideline 471, EU Method B.14 and EPA OTS 798.5265 in compliance with GLP.
Based on the results of a preliminary toxicity/range finding study, the substance was tested for mutagenic effects without and with metabolic activation at 5 doses from 61.7 to 5000 µg active ingredient/plate. An independent repetition of the experiments was performed with the same concentrations. In the preliminary experiment without and with metabolic activation performed on strain TA 100, a slight increase in the number of revertant counts was observed at the upper concentrations, followed by a reduction in the number of revertant colonies at the highest concentration. In the original experiment carried out without metabolic activation, treatment of strain TA 102 with the test substance led to a slight increase in the number of revertant counts at 185.2 to 1666.7 µg active ingredient/plate. In the original experiment with activation, a similar effect occurred on strain TA 100 at 1666.7 µg/plate and on strain TA 102 at 185.2 and 1666.7 µg active ingredient/plate. No effects were observed with the other strains. In the confirmatory experiment carried out without metabolic activation, treatment led to a slight increase in the number of revertant counts at 1666.7 µg active ingredient/plate. In the confirmatory experiment with activation, a similar effect occurred on strains TA 100 and TA 102 at the same concentration. Again, no effects were observed with the other strains. Overall, the test substance exerted only a marginal mutagenic action on strains S. typhimurium TA 100 and TA 102.
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