Registration Dossier
Registration Dossier
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 943-063-8 | CAS number: -
- 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:
- 07 July 2016 to 25 July 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
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:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Reaction mass of trisodium 4-[[ 4-chloro-6-[( 4-sulphonatophenyl)amino ]-1,3 ,5-triazin-2-yl ]amino ]-2-[[ 1-(2,5-dichloro-4-sulphonatophenyl)-4,5-dihydro-3-methyl-5-oxolH-pyrazol-4-yl]azo]benzenesulphonate and trisodium 4-[[4-chloro-6-[(3-sulphonatophenyl)amino ]-1,3,5-triazin-2-yl]amino ]-2-[[ 1-(2,5-dichloro-4-sulphonatophenyl)-4,5-dihydro-3-methyl-5-oxo-1 H-pyrazol-4-yl]azo] benzenesulphonate
- IUPAC Name:
- Reaction mass of trisodium 4-[[ 4-chloro-6-[( 4-sulphonatophenyl)amino ]-1,3 ,5-triazin-2-yl ]amino ]-2-[[ 1-(2,5-dichloro-4-sulphonatophenyl)-4,5-dihydro-3-methyl-5-oxolH-pyrazol-4-yl]azo]benzenesulphonate and trisodium 4-[[4-chloro-6-[(3-sulphonatophenyl)amino ]-1,3,5-triazin-2-yl]amino ]-2-[[ 1-(2,5-dichloro-4-sulphonatophenyl)-4,5-dihydro-3-methyl-5-oxo-1 H-pyrazol-4-yl]azo] benzenesulphonate
- Test material form:
- solid: particulate/powder
- Details on test material:
- - Appearance: Orange Powder
- Storage conditions: Room temperature
1
- Specific details on test material used for the study:
- TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
Test solution of the maximum concentration was prepared, which was fully stirred with mixer and dissolved after the test substance was weighed and the solvent was added. The lower doses were prepared by diluting stepwise from the test solution of the maximum concentration. The test was carried out with the weight converted by multiplying measured weight by 0.820 because the purity of the test substance was 82.0 wt %. Preparation of the test solution was carried out just prior to use under lamps with ultraviolet absorbent filter.
The amount of test substance and the volume of solvent used for the preparation of the test solution of the maximum concentration in each test were as follows.
Preliminary test [Prepared maximum concentration of test solution: 50 mg/mL]
- The amount of test substance 113.0 mg x 0.820 = 92.660 mg
- The volume of solvent (Water for injection) 1.853 mL
The first main test [Prepared maximum concentration of test solution: 50 mg/mL]
- The amount oftest substance 421.5 mg x 0.820 = 345.630 mg
- The volume of solvent (Water for injection) 6.913 mL
The second main test [Prepared maximum concentration of test solution: 50 mg/mL]
- The amount oftest substance 414.7 mg x 0.820 = 340.054 mg
- The volume of solvent (Water for injection) 6.801 mL
Method
- Target gene:
- - Histidine requirement in the Salmonella typhimurium strains (Histidine operon).
- Tryptophan requirement in the Escherichia coli strain (Tryptophan operon).
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: S. typhimurium TA100 was supplied by The Division of Mutagenesis, National Institute of Hygienic Sciences, Japan (The Division of Genetics and Mutagenesis, Biological Safety Research Center, National Institute of Health Sciences, Japan at present).
Other S. typhimurium TA strains were supplied by Dr. Ames, U.C. Berkeley, CA, U.S.A.
E. coli WP2 uvr A was supplied by Department of Molecular Oncology, Institute of Medical Science, University of Tokyo, Japan.
- Methods for maintenance in cell culture if applicable: The bacterial suspension and DMSO (spectrophotometric grade) were mixed in a ratio of 0.8 mL to 0.07 mL. The mixture was subdivided into 0.3 mL aliquots, and then frozen and stored at -85 to -80 °C.
- Charecterisation: The number of viable cell, amino acid requirement, UV sensitivity, rfa mutation, presence or absence of the drug resistance factor (R-factor plasmid) and positive control test (Dose-relation) were confirmed, and good strains were used as test strains.
- Pre-culture procedure: A bacterial suspension of each strain (20 μL of S. typhimurium TA strains, 5 μL of E. coli WP2 uvrA) was inoculated into an L-form culture tube (35 mL capacity) containing 10 mL of Nutrient Broth. This culture tube was left at 4 °C until starting incubation, and then incubated while shaking (100 rpm) in a water bath at 37 °C for 8 hours. After incubation, the optical density was measured and the number of viable cell was calculated by growth curve for each strain. The bacterial cultures were stored at room temperature until starting the test.
MEDIA USED
- Type and identity of media including CO2 concentration if applicable: Cultures were maintained in nutrient broth no. 2 supplied by Oxoid Limited. Top agar was bacto-agar (Difco Laboratories) and minimal glucose agar plates were Tesmedia AN (Oriental Yeast , Co. Ltd.).
- Properly maintained: Yes - Additional strain / cell type characteristics:
- other: Base-pair substitution type: Salmonella typhimurium TA100, TA1535 and Escherichia coli WP2 uvrA; Frame-shift type: Salmonella typhimurium T A98 and TA 1537
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix
- Test concentrations with justification for top dose:
- Preliminary test: 1.2, 4.9, 20, 78, 313, 1 250 and 5 000 μg/plate.
Definitive test: 313, 625, 1250, 2 500 and 5 000 μg/plate. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water for injection.
- Justification for choice of solvent/vehicle: Based on the information from the sponsor that the test substance was soluble at 100 g/L and more in water. Therefore water for injection was used as solvent for preparation.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Treated only with water that was used to prepare the test solution.
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- benzo(a)pyrene
- other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide, 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino Jacridine · 2HCl and 2-Aminoanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: Preincubation method
- Without metabolic activation: 0.5 mL of 0.1 M Na-phosphate buffer (pH 7.4) and 0.1 mL of each fresh bacterial culture were added to each tube containing 0.1 mL of the test solution or the negative control solution.
- With metabolic activation: 0.5 mL of the S9 mix was added to each tube instead of the 0.1 M Na-phosphate buffer.
DURATION
- Preincubation period: The mixture was pre-incubated in a water bath at 37 °C for 20 minutes while shaking horizontally, and then 2.0 mL of top agar were added to the mixture, and the contents of each tube were poured over the surface of the minimal glucose agar plate. And 0.1 mL of the positive control solution was carried out equally.
- Exposure duration: All plates were incubated at 37 °C for 48 hours, and the number of revertant colonies was counted. Afterwards, growth inhibition of the test strains was checked using a stereoscopic microscope.
NUMBER OF REPLICATIONS: One minimal glucose agar plate was used for each dose level in the preliminary test and three minimal glucose agar plates were used for each dose level in two main tests which were performed at the same doses.
- OTHER: For the sterility test, 0.1 mL of the test solution of the maximum concentration and 0.5 mL of the S9 mix were put into each tube, 2.0 mL of top agar were then added to the tube, and the contents of each tube were poured over the surface of the minimal glucose agar plate. These operations were conducted under lamps with ultraviolet absorbent filter.
As top agar, the 0.5 mM biotin-0.5 mM L-histidine solution and the 0.5 mM L-tryptophan solution were added to the soft agar solution (0.6 % Agar and 0.5 % NaCl) by volume of 1/10, for the S. typhimurium TA strains and the E. coli strain, respectively.
- Counting procedure: The number of revertant colonies was counted with a colony counter. - Rationale for test conditions:
- In the preliminary test, the growth inhibition by the test substance was not observed in any strains either with or without metabolic activation. And the precipitate of the test substance on the plates was not observed either with or without metabolic activation.
Therefore, as the highest dose level of the test substance in the main tests, the 5 000 μg/plate dose was selected for all strains both with and without metabolic activation. This highest dose was diluted 4 times (using a common ratio of 2) to provide a total of 5 dose levels. - Evaluation criteria:
- In the two main tests, if the number of revertant colonies on the test plates increased significantly in comparison with that on the control plates (based on twice as many as that of the negative control), and dose-response and reproducibility were also observed, the test substance was to be judged positive. The results at each concentration were demonstrated with the mean and the standard deviation.
Results and discussion
Test results
- Key result
- Species / strain:
- other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- In the two main tests, neither an increase in the number of revertant colonies (more than twice as many as that of the negative control) nor a dose-related response was observed at any doses in any strains of base-pair substitution type or frame-shift type, with or without metabolic activation.
The revertant colonies of the positive controls showed an increase of more than twice that of the negative controls and they were within limit of controls (mean ± 3SD) in historical data, indicating that this study was performed correctly.
From these results, mutagenicity of the test substance was judged negative. The growth inhibition of the test strains by the test substance was not observed. And the precipitate of the test substance on the plates was not observed either with or without metabolic activation.
In the sterility test on the test solution and the S9 mix, no growth of bacteria was observed.
Applicant's summary and conclusion
- Conclusions:
- Under the conditions of this study, the test material was considered to be non-mutagenic in the bacterial reverse mutation assay.
- Executive summary:
The potential of the test material to cause mutagenic effects in bacteria was assessed in accordance with the standardised guidelines OECD 471 and Japanese Guidelines for Screening Mutagenicity Testing Of Chemicals under GLP conditions.
Mutagenicity potential of the test material was assessed with Salmonella typhimurium TAl00, TA1535, TA98, TA1537 and Escherichia coli WP2 uvrA in to main tests. The dose range for the main test was determined from the preliminary test using the following seven dose levels: 1.2, 4.9, 20, 78, 313, 1 250 and 5 000 μg/plate. In the preliminary test, the growth inhibition by the test substance was not observed in any strains either with or without metabolic activation. And the precipitate of the test substance on the plates was not observed either with or without metabolic activation. Therefore, as the highest dose level of the test substance in the main tests, the 5 000 μg/plate dose was selected for all strains both with and without metabolic activation. This highest dose was diluted 4 times (using a common ratio of 2) to provide a total of 5 dose levels.
The main test concentrations were 313, 625, 1250, 2500 and 5 000 µg/plate. The revertant colonies of the positive controls showed an increase of more than twice that of the negative controls and they were within limit of controls (mean ± 3SD) in historical data, indicating that this study was performed correctly.
Neither an increase in the number of revertant colonies more than twice in comparison with that of the negative control, nor a dose-related response, was observed in any strains of base-pair substitution type or frame-shift type, with or without metabolic activation.
Under the conditions of this study, the test material was considered to be non-mutagenic in the bacterial reverse mutation assay.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
