Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
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.
Reaction mass of disodium 2-({4-methyl-2-[(4-{[5-methyl-1-(2-sulfonatophenyl)hexan-3-yl]amino}-9,10-dioxo-9,10-dihydroanthracen-1-yl)methyl]pentyl}amino)benzene-1-sulfonate,disodium 4-{5-methyl-3-[(4-{[5-methyl-1-(4-sulfonatophenyl)hexan-3-yl]amino}-9,10-dioxo-9,10-dihydroanthracen-1-yl)amino]hexyl}benzene-1-sulfonate anddisodium 4-{5-methyl-3-[(4-{[5-methyl-1-(2-sulfonatophenyl)hexan-3-yl]amino}-9,10-dioxo-9,10-dihydroanthracen-1-yl)amino]hexyl}benzene-1-sulfonate
EC number: 947-270-4 | 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:
- June 16th, 1998 to June 25th, 1998
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 998
- Report date:
- 1998
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- OECD Guideline For Testing Of Chemicals, 471 Bacterial Reverse Mutation Test Adopted: July 21st 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- EEC Directive 92/69, L 383 A, Annex B 14
EEC Directive 92/69, L 383 A, Annex B 13 - Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: U.S. EPA: 798.5265 The Salmonella typhimurium reverse mutation assay
- Version / remarks:
- U.S. EPA: 798.5265 The Salmonella typhimurium reverse mutation assay Fed. Reg. 50, Subpart F, September 1985
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: U.S. EPA: 798.5100 The Escherichia coli WP2 and WP2 uvrA reverse mutation assay
- Version / remarks:
- U.S. EPA: 798.5100 The Escherichia coli WP2 and WP2 uvrA reverse mutation assay Fed. Reg. 50, Subpart F, September 1985
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Disodium [(9,10-dihydro-9,10-dioxo-1,4-anthrylene)bis[imino[3-isopropylpropane-1,3-diyl]]]bis(benzenesulphonate)
- EC Number:
- 276-822-0
- EC Name:
- Disodium [(9,10-dihydro-9,10-dioxo-1,4-anthrylene)bis[imino[3-isopropylpropane-1,3-diyl]]]bis(benzenesulphonate)
- Cas Number:
- 72749-90-7
- Molecular formula:
- C40H46N2O8S2.2Na
- IUPAC Name:
- Disodium [(9,10-dihydro-9,10-dioxo-1,4-anthrylene)bis[imino[3-isopropylpropane-1,3-diyl]]]bis(benzenesulphonate)
- Test material form:
- solid: particulate/powder
- Details on test material:
- Acid Blue 221
Constituent 1
Method
- Target gene:
- histidine and tryptophan
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-Mix
- Test concentrations with justification for top dose:
- first experiment withand without metabolic activation: 50, 160, 500, 1600, 5000 μg/plate
second experiment withand without metabolic activation: 16, 50, 160, 500, 1600, 5000 μg/plate - Vehicle / solvent:
- Formulation of test compound
suspended in double-distilled water at appropriate concentrations immediately before use.
Formulation of reference compounds
sodium-azide dissolved in double-distilled water final concentration: 1.0 μg/plate for strain TA 100 and TA 1535
9-aminoacridine dissolved in DMSO final concentration: 50.0 μg/plate for strain TA 1537
2-nitrofluorene dissolved in DMSO final concentration: 2.5 μg/plate for strain TA 98
MNNG dissolved in DMSO final concentration: 4 μg/plate for strain WP2uvrA
2-aminoanthracene dissolved in DMSO final concentrations:
0.5 μg/plate for strain TA 98 and TA 100
1.0 μg/plate for strain TA 1535 and TA 1537
30.0 μg/plate for strain WP2uvrA
The frozen stock solutions of each compound were diluted progressively up to the final concentration on the day of treatment.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- other: 1-methyl-3-nitro-1-nitrosoguanidine (MNNG); 2-aminoanthracene (2-AA)
- Details on test system and experimental conditions:
- Assay procedure
The first mutation test was performed in both the presence and absence of S9-mix using all bacterial tester strains and a range of concentrations of the test substance.
Positive and negative controls as well as solvent controls were included in each test.
Triplicate plates were used.
The first mutation experiment also assessed the toxicity of the test substance by evaluation of the bacterial lawn in order to select a suitable range of dose levels for a second mutation test. The highest concentration was usually 50 mg/ml of the test substance in the chosen solvent, which provided a final concentration of 5000 μg/plate.
Further dilutions of 1600, 500, 160 and 50 μg/plate were used.
A reduced rate of spontaneously occurring colonies and visible thinning of the bacterial lawn were used as toxicity indicators. Thinning of the bacterial lawn was evaluated microscopically.
If the number of evaluable concentrations was less than 5, a repeat mutation experiment on the basis of toxicity results in the first test was performed.
If negative or equivocal results obtained, a second mutation experiment was performed on the basis of toxicity results in the first test as a pre-incubation test.
For mutagenicity testing top agar was prepared for the Salmonella strains by mixing 100 ml agar (0.6% (w/v) agar, 0.5% (w/v) NaCI) with 10 ml of a 0.5 mM histidinebiotin solution. With E. coli histidine was replaced by tryptophan (2.5 ml, 0.5 mM). The following ingredients were added (in the following order) to 2 ml of molten top agar at approx. 48 °C:
0.5 ml S9-mix (if required) or buffer
0.1 ml of an overnight nutrient broth culture of the bacterial tester strain
0.1 ml test compound solution (dissolved in double-distilled water)
Only for the second mutagenicity test this top-agar liquid was pre-incubated for approximately 20 minutes.
After mixing, the liquid was poured into a petri dish containing a 25 ml layer of minimal agar (1.5 % (w/v) agar, Vogel-Bonner E medium with 2% (w/v) glucose). After incubation for approximately 48 hours at approx. 37 °C in the dark, colonies (his+ and trp+ revertants) were counted with an automatic colony counter (Artec counter Model 880). The counter was calibrated for each test by comparison of manual count data of three control plates with automatic data of the colony counter.
A correction factor was determined to compensate for differences between manual and automatic count. This correction factor was used to automatically adjust the observed number of colonies on each plate to more accurately reflect the actual number of colonies present. - Rationale for test conditions:
- In accordance with test guidelines.
- Evaluation criteria:
- Criteria for a valid assay
The assay is considered valid if the following criteria are met:
-the solvent control data are within the laboratory's normal control range for the spontaneous mutant frequency
-the positive controls induced increases in the mutation frequency which were both statistically significant and within the laboratory's normal range
Criteria for a positive response
A test compound is classified as mutagenic if it has either of the following effects:
a) it produces at least a 2-fold increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control at complete bacterial backgrounc;tlawn
b) it induces a dose-related increase in the mean number of revertants per plate of at least one of the tester strains over the mean number of revertants per plate of the appropriate vehicle control in at least two to three concentrations of the test compound at complete bacterial background lawn.
If the test substance does not achieve either of the above criteria, it is considered to show no evidence of mutagenic activity in this system.
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
- Key result
- Species / strain:
- E. coli WP2 uvr A
- 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:
- Sterility checks and control plates
Sterility of S9-mix and the test compound were indicated by the absence of contamination on the test material and S9-mix sterility check plates. Control plates (background control and positive controls) gave the expected number of colonies, i.e. values were within the laboratory's historical control range.
Solubility and toxicity
The test compound was suspenden in double-distilled water and a stock solution of 50 mg/ml was prepared for the highest concentration, which provided a final concentration of 5000 μg/plate. Further dilutions of 1600, 500, 160, 50 and 16 μg/plate were used.
The test compound did not precipitate on the plates up to the highest investigated dose of 5000 μg/plate.
The test compound proved to be not toxic to the bacterial strains in the mutagenicity experiments.
Mutagenicity
In both independent mutation tests the number of colonies per plate with each strain as well as mean values of 3 plates are given.
The test compound did not cause a significant increase in the number of revertant colonies at any dose level with any of the tester strains either in the absence or in the presence of S9-mix in either mutation test. No dose-dependent effect was obtained.
All positive controls produced significant increases in the number of revertant colonies.
Thus, the sensitivity of the assay and the efficacy of the exogenous metabolic activation system were demonstrated.
Applicant's summary and conclusion
- Conclusions:
- The results lead to the conclusion that Acid Blue 221 is not mutagenic in these bacterial test systems either in the absence or in the presence of an exogenous metabolizing system.
The substance is therefore not classified in accordance with CLP criteria. - Executive summary:
Acis Blue 221 was tested for mutagenicity with the strains TA 100, TA 1535, TA 1537 and TA 98 of Salmonella typhimurium and Escherichia coli WP2uvrA.
Two independent mutagenicity studies were conducted, each in the absence and in the presence of a metabolizing system derived from a rat liver homogenate.
For both studies, the compound was dissolved in double-distilled water, and each bacterial strain was exposed to 5 dose levels in the first and 6 dose levels in the second experiment. Concentrations for both studies ranged from 50 to 5000 μg/plate.
Control plates without mutagen showed that the number of spontaneous revertant colonies was within the laboratory's historical control range. All the positive control compounds showed the expected increase in the number of revertant colonies.
Toxicity: In both mutagenicity experiments toxicity was not observed either with or without metabolic activation.
Mutagenicity: In the absence and in the presence of the metabolic activation system, Acid Blue 221 did not result in relevant increases in the number of revertants in any of the bacterial strains.
Summarizing, it can be stated that Acid Blue 221 was not mutagenic in this bacterial mutation test at any dose level in either the absence or in the presence of an exogenous metabolic activation.
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.