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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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.
EC number: 277-429-7 | CAS number: 73398-29-5
- 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:
- 28 June 2018 to 13 July 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Standards for Mutagenicity Tests using Microorganisms
- Version / remarks:
- (Notification No. 77, September 1, 1988 & partial revision: Notification No. 67, June 2, 1997, Ministry of Labour, Japan, Notification No. 120, December 25, 2000, Ministry of Labour, Japan and Notification No. 208, April 18, 2016, Ministry of Health, Labour and Welfare, Japan) and the "Amendment of the Reporting Form of the Results of the Mutagenicity Tests using Microorganisms" (Notification No. 653, September 29, 1997, Labour Standards Bureau, Ministry of Labour, Japan)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Guidelines for Toxicity Testings of New Chemical Substances
- Version / remarks:
- (Notification No. 7 of 0331 Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare, No. 5 of the Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry, & No. 110331009 of Environmental Policy Bureau, Ministry of the Environment, Japan, March 31, 2011 and partial revision: Notification No. 1 of 1221 Pharmaceutical Safety and Environmental Health Bureau, Ministry of Health, Labour and Welfare, No. 1 of the Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry, & No. 1512211 of Environmental Policy Bureau, Ministry of the Environment, Japan, December 21, 2015)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Disodium 2,2'-[(9,10-dihydro-9,10-dioxo-1,4-anthrylene)diimino]bis[3-bromo-5-butyltoluene-4-sulphonate]
- EC Number:
- 277-429-7
- EC Name:
- Disodium 2,2'-[(9,10-dihydro-9,10-dioxo-1,4-anthrylene)diimino]bis[3-bromo-5-butyltoluene-4-sulphonate]
- Cas Number:
- 73398-29-5
- Molecular formula:
- C36H34Br2N2O8S2.2Na
- IUPAC Name:
- disodium 2-bromo-3-({4-[(2-bromo-4-butyl-6-methyl-3-sulfonatophenyl)amino]-9,10-dioxo-9,10-dihydroanthracen-1-yl}amino)-6-butyl-4-methylbenzene-1-sulfonate
- Test material form:
- solid: particulate/powder
- Details on test material:
- - Appearance: Ultramarine blue powder
- Storage conditions: The test sample was stored in a dark storage place at room temperature.
Constituent 1
Method
- Target gene:
- - Histidine requirement in the Salmonella typhimurium strains (Histidine operon).
- Tryptophan requirement in the Escherichia coli strain (Tryptophan operon).
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: TA100: 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); April 15, 1982. TA1535, TA98 and TA1537: Dr. Ames, U.C. Berkeley, CA, U.S.A.; January 20, 1988 Department of Molecular Oncology, Institute of Medical Science, University of Tokyo, Japan; November 24, 1987
- Storage: 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.
- Characterisation: As characteristic test, 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.
- Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: Dr. Ames, U.C. Berkeley, CA, U.S.A.; January 20, 1988 Department of Molecular Oncology, Institute of Medical Science, University of Tokyo, Japan; November 24, 1987
- Storage: 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.
- Characterisation: As characteristic test, 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.
- 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, 1250 and 5000 µg/plate
- In the preliminary test, the growth inhibition by the test material was observed at 313 µg/plate and more in S. typhimurium TA 1537 without metabolic activation, and at 1250 µg/plate and more in S. typhimurium TA100 and TA98 without metabolic activation and S. typhimurium TA1537 with metabolic activation, and at 5000 µg/plate in S. typhimurium TAl100 and TA98 with metabolic activation. And the precipitate of the test material on the plates was not observed either with or without metabolic activation.
- Therefore, as the highest dose level of the test material in the main tests, the 313 µg/plate dose was selected for S. typhimurium TA 1537 without metabolic activation, and the 1250 µg/plate dose was selected for S. typhimurium TA 100 and T A98 without metabolic activation and S. typhimurium TA1537 with metabolic activation, and the 5000 µg/plate dose was selected for S. typhimurium TA100 and TA98 with metabolic activation, and these highest doses were diluted 5 times (using a common ratio of 2) to provide a total of 6 dose levels. And the 5000 µg/plate dose was selected for S.typhimurium TA1535 and E.coli WP2 uvrA both with and without metabolic activation, and this highest dose was diluted 4 times (using a common ratio of 2) to provide a total of 5 dose levels.
- First and Second Main Test: 10, 20, 39, 78, 156, 313, 625, 1250, 2500 and 5000 µg/plate (-S9 mix)
- First and Second Main Test: 39, 78, 156, 313, 625, 1250, 2500 and 5000 µg/plate (+S9 mix) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Based on the information from the sponsor that the solubility of the test material in water was 23 g/L and less, the solubility test was performed with DMSO. The test material was dissolved at 50 mg/mL in DMSO. Therefore DMSO was used as solvent for preparation.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- 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 (AF-2), 2-Methoxy-6-chloro-9-[3-(2-chloroethyl )aminopropy lamino ]acridine·2HCl (ICR-191) and 2-Aminoanthracene (2AA)
- Details on test system and experimental conditions:
- PREPARATION OF THE TEST SOLUTION
- Test solution of the maximum concentration was prepared, which was fully stirred with mixer and dissolved after the test material 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 0.900 because the purity of the test material was 90.0 wt %. Preparation of the test solution was carried out just prior to use under lamps with ultraviolet absorbent filter.
METHOD OF APPLICATION: preincubation
DURATION
- Pre-culture period 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. The number of bacteria in the culture was estimated from the optical density at the end of the pre-culture.
- Pre-incubation method test procedure: For the tests 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. For the tests with metabolic activation, 0.5 mL of the S9 mix was added to each tube instead of the 0.1 M Na-phosphate buffer. 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. 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. 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. 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.
COUNTING PROCEDURE
- The number of revertant colonies was counted visually due to the colour of the test material on the plates. But the revertant colonies of positive controls were counted with a colony counter. - 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 material was to be judged positive. The results at each concentration were demonstrated with the mean and the standard deviation.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium, other: TA98, TA100, TA1535 & TA1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Except for TA1535
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- 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:
- not applicable
- 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 ± 3 S.D.) in historical data, indicating that this study was performed correctly.
- From these results, mutagenicity of the test material was judged negative.
- Precipitate of the test material on the plates was not observed either with or without metabolic activation.
Any other information on results incl. tables
Table 1: Summary of Main Experiment 1
± S9 Mix |
Concentration (µg/plate) |
Mean number of colonies/plate |
||||
Base-pair Substitution Type |
Frameshift Type |
|||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||
- |
Solvent 10 20 39 78 156 313 625 1250 2500 5000 |
95 - - 106 98 110 100 98 89* - - |
9 - - - - - 10 8 7 8 7 |
28 - - - - - 25 29 26 27 27 |
25 - - 25 28 22 22 22 19* - - |
11 11 8 10 10 9* 8* - - - - |
+ |
Solvent 39 78 156 313 625 1250 2500 5000 |
111 - - 104 111 111 104 93 87* |
11 - - - 8 9 9 8 8 |
32 - - - 31 32 29 32 29 |
36 - - 39 32 30 28 27* 18* |
18 17 19 17 15 13* 11* - - |
Positive Controls |
||||||
- |
Name |
AF-2 |
NaN3 |
AF-2 |
AF-2 |
ICR-191 |
Concentration (µg/plate) |
0.01 |
0.5 |
0.01 |
0.1 |
1.0 |
|
Mean no. colonies/plate |
528 |
394 |
150 |
525 |
1318 |
|
+ |
Name |
B[a]P |
2AA |
2AA |
B[a]P |
B[a]P |
Concentration (µg/plate) |
5.0 |
2.0 |
10.0 |
5.0 |
5.0 |
|
Mean no. colonies/plate |
694 |
220 |
306 |
195 |
66 |
2AA = 2-aminoanthracene
B[a]P = benzo(a)pyrene
NaN3 = sodium azide
AF-2 = 2-(2 -furyl)-3-(5-nitro-2-furyl)acrylamide
ICR-191 = 2 -methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino]acridine.2HCl
* = Growth inhibition of the tested bacterium by the test material was observed.
Table 2: Summary of Main Experiment 2
± S9 Mix |
Concentration (µg/plate) |
Mean number of colonies/plate |
||||
Base-pair Substitution Type |
Frameshift Type |
|||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||
- |
Solvent 10 20 39 78 156 313 625 1250 2500 5000 |
104 - - 113 112 114 114 103 112* - - |
12 - - - - - 11 9 8 8 8 |
26 - - - - - 26 22 25 22 23 |
29 - - 27 32 28 22 23 17* - - |
9 7 8 9 8 9* 8* - - - - |
+ |
Solvent 39 78 156 313 625 1250 2500 5000 |
128 - - 124 126 118 107 106 97* |
8 - - - 8 9 9 7 7 |
30 - - - 29 27 23 26 23 |
33 - - 34 32 30 25 23* 18* |
17 16 17 15 15 9* 8* - - |
Positive Controls |
||||||
- |
Name |
AF-2 |
NaN3 |
AF-2 |
AF-2 |
ICR-191 |
Concentration (µg/plate) |
0.01 |
0.5 |
0.01 |
0.1 |
1.0 |
|
Mean no. colonies/plate |
517 |
411 |
126 |
535 |
1273 |
|
+ |
Name |
B[a]P |
2AA |
2AA |
B[a]P |
B[a]P |
Concentration (µg/plate) |
5.0 |
2.0 |
10.0 |
5.0 |
5.0 |
|
Mean no. colonies/plate |
706 |
223 |
323 |
169 |
57 |
2AA = 2-aminoanthracene
B[a]P = benzo(a)pyrene
NaN3 = sodium azide
AF-2 = 2-(2 -furyl)-3-(5-nitro-2-furyl)acrylamide
ICR-191 = 2 -methoxy-6-chloro-9-[3-(2-chloroethyl)aminopropylamino]acridine.2HCl
* = Growth inhibition of the tested bacterium by the test material was observed.
Applicant's summary and conclusion
- Conclusions:
- Under the conditions of this study, the test material was not mutagenic in the bacterial reverse mutation assay.
- Executive summary:
The genetic toxicity of the test material was investigated in accordance with the standardised guideline OECD 471 and other Japanese guidelines, under GLP conditions.
The mutagenicity potential of the test material was assessed with Salmonella typhimurium strains TA100, TA1535, TA98, TA1537 and Escherichia coli strain WP2 uvrA in the bacterial reverse mutation assay.
Bacteria were exposed to the test material using a pre-incubation method both in the presence and absence of metabolic activation in the form of S9 mix. All plates were incubated at 37 °C for 48 hours, and the number of revertant colonies were counted. Positive and solvent controls were also included in the study.
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. In the sterility test on the test solution and the S9 mix, no growth of bacteria was observed.
Under the conditions of this study, the test material was not 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.