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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 249-352-9 | CAS number: 28983-56-4
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Gene mutation toxicity study was performed by Lynnette R. Fergusonet al (Mutation Research, 1988) to determine the mutagenic nature of target substance Methyl blue ;IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4)using Salmonella typhimurium strains. In Genetic toxicity in vitro study, Methyl Blue was assessed for its possible mutagenic potential. For this purpose AMES assay was performed in Salmonella typhimurium TA1537, TA1598 and TA100 by plate-incorporation assay. The test substance was exposed at the concentration of than 100µl of 50% ethanol, or 50% ethanol alone as negative control. Plates were allowed to harden, and then incubated for 3 days at 37°C before scoring colonies for reversion to Histidine independence. A dose range of each compound was tested, and each experimental point performed in triplicate on at leasttwo separate occasions. No mutagenic effects were observed at this dose .Therefore Methyl Blue was considered to be non mutagenic for AMES test. Hence the substance cannot be classified as gene mutant in vitro.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Justification for type of information:
- Data is from publication.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Principles of method if other than guideline:
- To evaluate the mutagenic potential of Methyl Blue in Salmonella typhimurium TA1537, TA1598 and TA100 by AMES assay.
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- - Name of test material :Methyl Blue
- Molecular formula : C37H26N3Na2O9S3
- Molecular weight : 800.8182 g/mol
- Smiles notation : c1cc(ccc1C(=C2C=CC(=[NH+]c3ccc (cc3)S(=O) (=O)[O-])C=C2)c4ccc(cc4)Nc5ccc(cc5)S (=O)(=O)[O-])Nc6ccc(cc6)S(=O)(=O)O.[Na+].[Na+]
- InChl : 1S/C37H29N3O9S3.2Na/c41-50(42,43)34-19-13-31(14-20-34)38-28-7-1-25(2-8-28)37(26-3-9-29(10-4-26)39-32-15-21-35(22-16-32)51(44,45 46)27-5-11-30(12-6-27)40-33-17-23-36(24-18-33)52(47,48)49;;/h1-24,38-39H,(H,41,42,43)(H,44,45,46)(H,47,48,49);;/q;2*+1/p-1
- Substance type: Organic
- Physical state: Solid - Target gene:
- Histidine
- Species / strain / cell type:
- S. typhimurium, other: TA1537, TA1598 and TA100
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- not specified
- Metabolic activation:
- not specified
- Test concentrations with justification for top dose:
- 100 µl
- Vehicle / solvent:
- 50% of ethanol
- Untreated negative controls:
- yes
- Remarks:
- ethanol
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol
- True negative controls:
- not specified
- Positive controls:
- no
- Details on test system and experimental conditions:
- Details on test system and conditions
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 3days
NUMBER OF REPLICATIONS: triplicate
OTHER: Reversion characteristics of each strain were tested in each experiment using the disc method of Zieger et al. (1982). The background number of revertants was 8 ± 3 for TA1537, 34 ±7 for TA98, and 171 ± 10 for TAI00. - Rationale for test conditions:
- Not specified
- Evaluation criteria:
- Mutagenicity is expressed as revertants colonies/µg compound added to the plate, and is derived from the slope of the corresponding regression equation.
- Statistics:
- Mutagenicity data were analyzed according to the method of Moore and Felton (1983). A regression line was fitted for the number of mutants versus
drug concentration for the linear part of the curve. Mutagenicity was scored as negative if the regression coefficient was non-significant. To determine whether verapamil had a significant effect on mutagenicity, the slopes ( +__ SD) of the corresponding regression lines with and without verapamil were determined, and the significance of the difference was calculated using Student's t test. - Key result
- Species / strain:
- S. typhimurium, other: TA1537, TA1598 and TA100
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Additional information on results:
- All mutagenicity data, calculated as revertant colonies per /µg of added drug in the presence or absence of verapamil.
- Remarks on result:
- other: NO mutagenic effct were observerd
- Conclusions:
- Methyl Blue (28983-56-4) was evaluated for its mutagenic potential in Salmonella typhimurium TA1537, TA1598 and TA100. The test result was considered to be negative for the test.
- Executive summary:
In Genetic toxicity in vitro study, Methyl Blue was assessed for its possible mutagenic potential. For this purpose AMES assay was performed in Salmonella typhimurium TA1537, TA1598 and TA100 by plate-incorporation assay. The test substance was exposed at the concentration of than 100µl of 50% ethanol, or 50% ethanol alone as negative control. Plates were allowed to harden, and then incubated for 3 days at 37°C before scoring colonies for reversion to Histidine independence. A dose range of each compound was tested, and each experimental point performed in triplicate on at leasttwo separate occasions. No mutagenic effect were observed at this dose .Therefore Methyl Blue was considered to be non mutagenic for AMES test. Hence the substance cannot be classified as gene mutant in vitro.
Reference
Sr no |
Compound |
Mutagenicitybin bacterial strainc |
||
|
|
TA1537 |
TA98 |
TA100 |
1 |
Methyl Blue |
0 |
0 |
0 |
b Mutagenicity is expressed as revertant colonies/tzg compound added to the plate, and is derived from the slope of the corresponding
regression equation,.
c Bacterial strains are described in Maron and Ames (1983).
d Statistical significance of the differences: **p < 0.001; *p < 0.05.
e A value of 0 means that the correlation coefficient of the regression equation is not statistically significant.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Genetic toxicity In-vitro
Various experimental studies were reviewed to determine the mutagenic nature of target substance Methyl blue ; IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4). The studies are as mentioned below:
Gene mutation toxicity study was performed by Lynnette R. Fergusonet al (Mutation Research, 1988) to determine the mutagenic nature of target substance Methyl blue ;IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4)using Salmonella typhimurium strains. In Genetic toxicity in vitro study, Methyl Blue was assessed for its possible mutagenic potential. For this purpose AMES assay was performed in Salmonella typhimurium TA1537, TA1598 and TA100 by plate-incorporation assay. The test substance was exposed at the concentration of than 100µl of 50% ethanol, or 50% ethanol alone as negative control. Plates were allowed to harden, and then incubated for 3 days at 37°C before scoring colonies for reversion to Histidine independence. A dose range of each compound was tested, and each experimental point performed in triplicate on at leasttwo separate occasions. No mutagenic effects were observed at this dose .Therefore Methyl Blue was considered to be non mutagenic for AMES test. Hence the substance cannot be classified as gene mutant in vitro.
Supported by an experimental study conducted by William Au (Environmental Mutagenesis, 1979) to determine the mutagenic nature of target substance Methyl blue IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4) . In Genetic toxicity in vitro was performed study,Methyl blue wasassessed for its possible clastogenic potential. For this purpose Invitro chromosome breakage assay was performed in Chinese hamster ovary cell line (CHO). The test material was exposed at the concentration of 1-20 µM in the presence and absence of metabolic activation. The 50 well-spread metaphases were scored for chromosome aberration for each experimental point. The average number of breaks per metaphase was calculated and was used for comparing the clastogenic activities of different compounds. The average number of chromosome breaks ranged from 0 to 0.16 per cell for the water-treated control. No chromosome breaks were observed for target substance compare to control. No statically significant value was also observed for test substance. There to for Methyl blue was considered to be non clastogenic in Chinese hamster ovary cell line (CHO) by In vitro chromosome breakage assay in the presence and absence of metabolic activation. Hence the substance cannot be classified as gene mutant in vitro.
It is further supported by another experimental study conducted by Zev Leifer 2et al(Mutation Research,1981) ) to determine the mutagenic nature of target substance Methyl blue IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4). In Genetic toxicity in vitro was performed study, Methyl blue was assessed for its possible mutagenic potential. For this purpose REPAIR-DEFICIENCY ASSAYS was performed in bacteria E.coli and Bacillus SubtilisH17A. The test was performed by Diffusion tests and Suspension method. The zone of growth inhibition in the agar diffusion procedure and viability in suspension method was observed for test substance. Due to lack of zone of growth inhibition in the agar diffusion procedure and no viability in suspension method were taken as a No Test result for test substance. Therefore the test result can be considered as ambiguous.
Based on the experimental data , available for the target chemical, Methyl blue ; IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4)does not induce gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.
Justification for classification or non-classification
Based on above annotation for the target chemical, Methyl blue ; IUPAC name: Disodium [[4-[bis[4-[(sulphonatophenyl)amino] phenyl]methylene] cyclohexa-2,5-dien-1-ylidene]amino]ben(28983-56-4)does not induce gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.
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.