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A mixture of isomers of: 1,1'-[(3,5(or 2,4 or 4,6 or 2,6)-dihydroxy-o(or m or p)-phenylene)bis(azo-meta-phenyleneazo{1-[3-(dimethylamino)propyl]-1,2-dihydro-6-hydroxy-4-methyl-2-oxopyridine-5,3-diyl})]dipyridinium-dichloride-dihydrochloride; 1-(1-[3-(dimethylamino)propyl]-5-{3-[x-(4-{1-[3-(dimethylamino)propyl]-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-5-pyridinio-3-pyridylazo}phenylazo)-2,4(or 2,6 or 3,5 or 4,6)-dihydroxyphenylazo]phenylazo}-1,2-dihydro-6-hydroxy-4-methyl-2-oxo-3-pyridyl)pyridinium-dichloride-dihydrochloride (where x is variable)
EC number: 404-540-1 | CAS number: 159405-95-5 BRAUN HM 2763; BROWN HM 2763; BRUN HM 2763; BRUNO HM 2763
- 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
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- 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:
- 1989-03-09 to 1989-04-14
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 989
- Report date:
- 1989
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1983
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- 1984
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- A mixture of isomers of: 1,1'-[(3,5(or 2,4 or 4,6 or 2,6)-dihydroxy-o(or m or p)-phenylene)bis(azo-meta-phenyleneazo{1-[3-(dimethylamino)propyl]-1,2-dihydro-6-hydroxy-4-methyl-2-oxopyridine-5,3-diyl})]dipyridinium-dichloride-dihydrochloride; 1-(1-[3-(dimethylamino)propyl]-5-{3-[x-(4-{1-[3-(dimethylamino)propyl]-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-5-pyridinio-3-pyridylazo}phenylazo)-2,4(or 2,6 or 3,5 or 4,6)-dihydroxyphenylazo]phenylazo}-1,2-dihydro-6-hydroxy-4-methyl-2-oxo-3-pyridyl)pyridinium-dichloride-dihydrochloride (where x is variable)
- EC Number:
- 404-540-1
- EC Name:
- A mixture of isomers of: 1,1'-[(3,5(or 2,4 or 4,6 or 2,6)-dihydroxy-o(or m or p)-phenylene)bis(azo-meta-phenyleneazo{1-[3-(dimethylamino)propyl]-1,2-dihydro-6-hydroxy-4-methyl-2-oxopyridine-5,3-diyl})]dipyridinium-dichloride-dihydrochloride; 1-(1-[3-(dimethylamino)propyl]-5-{3-[x-(4-{1-[3-(dimethylamino)propyl]-1,6-dihydro-2-hydroxy-4-methyl-6-oxo-5-pyridinio-3-pyridylazo}phenylazo)-2,4(or 2,6 or 3,5 or 4,6)-dihydroxyphenylazo]phenylazo}-1,2-dihydro-6-hydroxy-4-methyl-2-oxo-3-pyridyl)pyridinium-dichloride-dihydrochloride (where x is variable)
- Cas Number:
- 159405-95-5
- Molecular formula:
- C50H56Cl4N14O6
- IUPAC Name:
- 1'-[3-(dimethylamino)propyl]-5'-(2-{3-[2-(4-{2-[3-(2-{1'-[3-(dimethylamino)propyl]-6'-hydroxy-4'-methyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium-5'-yl}diazen-1-yl)phenyl]diazen-1-yl}-2,6-dihydroxyphenyl)diazen-1-yl]phenyl}diazen-1-yl)-6'-hydroxy-4'-methyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium 1'-[3-(dimethylamino)propyl]-5'-(2-{4-[2-(4-{2-[4-(2-{1'-[3-(dimethylamino)propyl]-6'-hydroxy-4'-methyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium-5'-yl}diazen-1-yl)phenyl]diazen-1-yl}-2,6-dihydroxyphenyl)diazen-1-yl]phenyl}diazen-1-yl)-6'-hydroxy-4'-methyl-2'-oxo-1',2'-dihydro-1λ⁵-[1,3'-bipyridin]-1-ylium tetrahydrochloride tetrachloride
- Test material form:
- solid
Constituent 1
Method
- Target gene:
- The objective of this study was to evaluate the test substance for its ability to induce reverse mutations in a gene of histidine-requiring Salmonella Iyphimurium bacterial strains to produce histidine-independent strains of these micro-organisms.
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: rfa: deep rough (defective lipopolysaccharide cellcoat); gal: mutation in the galactose metabolism; chi: mutation in nitrate reductase; bio: defective biotin synthesis; uvrB: loss of the excision repair system (deletion of the ultravioletrepair B gene)
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor induced rat liver S9
- Test concentrations with justification for top dose:
- Concentration range in the main test (with metabolic activation): 3.3, 10, 33.3, 100, 333 µg/plate
Concentration range in the main test (without metabolic activation): 1, 3.3, 10, 33.3, 100 µg/plate - Vehicle / solvent:
- Solvent: deionised (Milli-Q) water
Controlsopen allclose all
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- sodium azide
- methylmethanesulfonate
- other: daunomycine
- Remarks:
- without S9-mix
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- with S9-mix
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (direct plate incorporation)
DURATION
- Preincubation period: no
- Exposure duration: 48 h
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): 48 h
- Fixation time (start of exposure up to fixation or harvest of cells): 48 h
SELECTION AGENT (mutation assays): histidine
NUMBER OF REPLICATIONS: 3
DETERMINATION OF CYTOTOXICITY
The survival of the TA100 culture is determined by comparing the number of colonies on the plates containing the test substance with those on the solvent control plate.
In the absence of S9-mix the survival of strain TA100 is severely reduced at test substance concentrations from 333 µg/plate upwards. Both in the absence and presence of S9-mix the survival is eliminated from 1000 µg/plate upwards. Based on these data, the test substance was tested up to a concentration of 100 µg/plate in the absence of S9-mix and up to 333 µg/plate in the presence of S9-mix. - Evaluation criteria:
- A test substance was considered negative (not mutagenic) in the Ames test if:
a) The total number of revertants in any tester strain at any concentration was not greater than two times the solvent control value, with or without metabolic activation.
b) The negative response should be reproducible in at least one independently repeated experiment.
A test substance was considered positive (mutagenic)in the Ames test if:
a) It induced at least a 2-fold, dose related increase in the number of revertants with respect to the number induced by the solvent control in any of the tester strains, either with or without metabolic activation. However, any mean plate count of less than 20 was considered to be not significant. If the test substance showed in the first test only a positive response at one or two concentrations, the assay was repeated with doses just below and exceeding those showing positive effects in the first test.
b) The positive response should be reproducible in at least one independently repeated experiment. - Statistics:
- see "data evaluation criteria"
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- other: not tested; only 4 strains.
- Cytotoxicity / choice of top concentrations:
- other: not tested; only 4 strains.
- Vehicle controls validity:
- not applicable
- Positive controls validity:
- not applicable
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- without
- Genotoxicity:
- not determined
- Remarks:
- range-finding test
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ≥ 333 µg/plate
- Vehicle controls validity:
- valid
- Positive controls validity:
- not applicable
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with
- Genotoxicity:
- not determined
- Remarks:
- range-finding test
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ≥ 1000 µg/plate
- Vehicle controls validity:
- not specified
- Positive controls validity:
- not applicable
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data
- Effects of osmolality: no data
- Evaporation from medium: no data
- Water solubility: no effects
- Precipitation: no
RANGE-FINDING/SCREENING STUDIES:
Selection of an adequate range of doses was based on a preliminary toxicity test with strain TA100, both with and without S9-mix. Nine concentrations have been tested in duplicate for toxicity. The highest concentration of test article used in the subsequent mutagenesis assay was that which gave a reduced survival on the non-selective plates. If no toxicity was observed, the highest dose level used in the mutagenesis assay was 5 mg/plate unless the test article exhibited limited solubility or was not uniformly dispersible in the solvent of choice.
COMPARISON WITH HISTORICAL CONTROL DATA:
The negative and strain-specific positive control values fell within the laboratory's background historical ranges indicating that the test conditions were optimal and that the metabolic activation system functioned properly.
Any other information on results incl. tables
Preliminary toxicity determination in TA 100
conc. µg/plate | viable counts/plate (duplicate plates) | |||
without S9 mix | with S9-mix | |||
control | 420 | 465 | 490 | 500 |
1.0 | 317 | 385 | 357 | 365 |
3.3 | 418 | 350 | 366 | 348 |
10.0 | 398 | 377 | 319 | 345 |
33.3 | 358 | 318 | 413 | 372 |
100 | 298 | 199 | 318 | 353 |
333 | 87 | 98 | 218 | 225 |
1000 | 0 | 0 | 0 | 0 |
3330 | 0 | 0 | 0 | 0 |
5000 | 0 | 0 | 0 | 0 |
Experiment 1
conc. µg/plate | without S9-mix | |||
TA1535 | TA1537 | TA98 | TA100 | |
control | 13±2 | 18±4 | 32±2 | 120±11 |
1.0 | 10±4 | 20±6 | 28±4 | 121±18 |
3.3 | 10±3 | 17±6 | 27±2 | 102±12 |
10.0 | 6±4 | 15±2 | 26±7 | 112±15 |
33.3 | 11±3 | 22±4 | 35±6 | 121±3 |
100 | 8±1 | 20±6 | 22±4 | 121±15 |
positive control | 200±13 | 477±72 | 503±130 | 576±267 |
with S9-mix | ||||
control | 8±2 | 11±4 | 26±4 | 114±13 |
3.3 | 6±1 | 15±3 | 22±1 | 124±7 |
10.0 | 7±2 | 15±4 | 23±2 | 11±10 |
33.3 | 8±5 | 10±1 | 26±9 | 123±7 |
100 | 10±2 | 16±6 | 26±3 | 125±3 |
333.3 | 10±3 | 13±3 | 30±7 | 112±3 |
positive control | 189±20 | 414±14 | 599±48 | 730±84 |
Experiment 2
conc. µg/plate | without S9-mix | |||
TA1535 | TA1537 | TA98 | TA100 | |
control | 8±5 | 13±4 | 28±6 | 105±20 |
1.0 | 12±3 | 15±3 | 31±11 | 109±11 |
3.3 | 12±5 | 15±1 | 36±10 | 114±12 |
10.0 | 9±3 | 15±4 | 30±10 | 110±5 |
33.3 | 10±7 | 16±4 | 22±2 | 108±7 |
100 | 11±2 | 14±1 | 28±4 | 126±8 |
positive control | 201±6 | 477±72 | 1103±70 | 586±54 |
with S9-mix | ||||
control | 9±2 | 7±1 | 34±2 | 122±2 |
3.3 | 15±1 | 13±2 | 38±6 | 135±11 |
10.0 | 9±1 | 8±5 | 26±4 | 126±9 |
33.3 | 14±1 | 8±3 | 35±8 | 125±6 |
100 | 13±3 | 11±1 | 37±5 | 132±3 |
333.3 | 8±4 | 10±2 | 44±3 | 139±12 |
positive control | 161±34 | 588±76 | 705±29 | 815±7 |
Applicant's summary and conclusion
- Conclusions:
- Negative: test substance can be considered as not mutagenic under test conditions of the Ames Salmonella/microsome assay.
- Executive summary:
Aim of the study was to evaluate the test substance for its ability to induce reverse mutations in a gene of histidine-requiring Salmonella typhimurium bacterial strains to produce histidine-independent strains of these micro-organisms. The study followed OECD guideline 471.
The substance was tested in the Ames Salmonella/microsome plate test up to 100 µg/plate in the absence of S9-mix and up to 333 µg/plate in the presence of S9-mix (dose range as result of a preliminary range finding study).
All bacterial strains showed negative responses over the entire dose range of the test substance, i.e. no dose related, two-fold, increase in the number of revertants in two independently repeated experiments. The negative and strain-specific positive control values fell within our laboratory background historical ranges indicating that the test conditions were optimal and that the metabolic activation system functioned properly.
Based on these findings, the test substance did not induce a dose-related increase in the number of revertant (His+) colonies in each of the four tester strains (TA1535; TA1537; TA98 and TA100). These results were confirmed in an independently repeated experiment. The test substance can, thus, be considered as not mutagenic under test conditions.
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