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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 249-008-8 | CAS number: 28407-37-6
- 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
![](https://chesar.echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from NTP report
Data source
Reference
- Reference Type:
- review article or handbook
- Title:
- Toxicology And Carcinogenesis Studies Of Direct Blue 218 (Cas No. 28407-37-6) In F344/N Rats And B6C3F1 Mice (Feed Studies)
- Author:
- U.S. Department Of Health And Human Services
- Year:
- 1 994
- Bibliographic source:
- National Toxicology Program, NTP TR 430, NIH Publication No. 94-3161, 1994
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- 14 days repeated dose toxicity study was conducted to determine the toxic nature of C.I. Direct blue 218
- GLP compliance:
- not specified
- Limit test:
- no
Test material
- Reference substance name:
- Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)
- EC Number:
- 249-008-8
- EC Name:
- Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)
- Cas Number:
- 28407-37-6
- Molecular formula:
- C32H16Cu2N6O16S4.4Na
- IUPAC Name:
- Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)
- Test material form:
- other: amorphous powder
- Details on test material:
- Name of the test chemical: Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)
Common Name: C.I Direct Blue 218
IUPAC name: tetrasodium (3E)-5-amino-3-{2-[4-(4-{2-[(2E)-8-amino-1-oxo-3,6-disulfonato-1,2-dihydronaphthalen-2-ylidene]hydrazin-1-yl}-3- hydroxyphenyl)-2-hydroxyphenyl]hydrazin-1-ylidene}-4-oxo-3,4-dihydronaphthalene-2,7-disulfonate dicopper
Molecular Formula: C32H20Cu2N6Na4O16S4
Molecular Weight: 1087.84 g/mol
SMILES Notation: c12c3c(c(S(=O)(=O)[O-])cc1cc(S(=O)(=O)[O])cc2N)N=Nc1ccc(cc1O[Cu]O3)c1cc2c(N=Nc3c(cc4c(c3O[Cu]O2)c(cc(c4)S(=O) (=O)[O-])N)S(=O)(=O)[O-])cc1.[Na+].[Na+].[Na+].[Na+]
InChI: 1S/C32H24N6O16S4.2Cu.4Na/c33-19-11-17(55(43,44)45)5-15-9-25(57(49,50)51)29(31(41)27(15)19)37-35-21-3-1-13(7-23(21)39) 14-2-4-22(24(40)8-14)36-38-30-26(58(52,53)54)10-16-6-18(56(46,47)48)12-20(34)28(16)32(30)42;;;;;;/h1-12,39-42H,33-34H2,(H,43,44,45)(H,46,47,48)(H,49,50,51)(H,52,53,54);;;;;;/q;2*+2;4*+1/p-8/b37-35-,38-36-;;;;;;
Substance Type: Organic
Physical State: Solid Deep purple to dark blue amorphous powder
Constituent 1
- Specific details on test material used for the study:
- - Name of the test chemical: Tetrasodium [μ-[[3,3'-[(3,3'-dihydroxy[1,1'-biphenyl]-4,4'-diyl)bis(azo)]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonato]](8-)]]dicuprate(4-)
- Common Name: C.I Direct Blue 218
- IUPAC name: tetrasodium (3E)-5-amino-3-{2-[4-(4-{2-[(2E)-8-amino-1-oxo-3,6-disulfonato-1,2-dihydronaphthalen-2-ylidene]hydrazin-1-yl}-3- hydroxyphenyl)-2-hydroxyphenyl]hydrazin-1-ylidene}-4-oxo-3,4-dihydronaphthalene-2,7-disulfonate dicopper
- Molecular Formula: C32H20Cu2N6Na4O16S4
- Molecular Weight: 1087.84 g/mol
- SMILES Notation: c12c3c(c(S(=O)(=O)[O-])cc1cc(S(=O)(=O)[O])cc2N)N=Nc1ccc(cc1O[Cu]O3)c1cc2c(N=Nc3c(cc4c(c3O[Cu]O2)c(cc(c4)S(=O) (=O)[O-])N)S(=O)(=O)[O-])cc1.[Na+].[Na+].[Na+].[Na+]
InChI: 1S/C32H24N6O16S4.2Cu.4Na/c33-19-11-17(55(43,44)45)5-15-9-25(57(49,50)51)29(31(41)27(15)19)37-35-21-3-1-13(7-23(21)39) 14-2-4-22(24(40)8-14)36-38-30-26(58(52,53)54)10-16-6-18(56(46,47)48)12-20(34)28(16)32(30)42;;;;;;/h1-12,39-42H,33-34H2,(H,43,44,45)(H,46,47,48)(H,49,50,51)(H,52,53,54);;;;;;/q;2*+2;4*+1/p-8/b37-35-,38-36-;;;;;;
- Substance Type: Organic
- Physical State: Dark blue solid
- Impurities (identity and concentrations): More than 9 minor components (>1%) were present in the dye and although it was not possible 10 determine the definite structure of these impurities, the reducible azo bond appeared 10 be present in the majoriIy of the impurities.
Test animals
- Species:
- mouse
- Strain:
- B6C3F1
- Remarks:
- F344/N
- Details on species / strain selection:
- No data
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Breeding Laboratories (Portage, MI)
- Age at study initiation: 50 to 57 days old
- Weight at study initiation:
- Fasting period before study: No data
- Housing: Animals were housed 5/cage in polycarbonate cages (changed twice weekly) with heat-treated hardwood chips bedding (changes twice weekly) and reemay spun bonded polyester cage filters (changed once every 2 weeks) having Stainless steel racks (changed once/2 weeks)
- Diet (e.g. ad libitum): NIH-D7 open-formula mash diet ad libitum
- Water (e.g. ad libitum): Water was provided through Automatic watering system ad libitum
- Acclimation period: 15 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22˚C
- Humidity (%): 68%
- Air changes (per hr): 12 changes/hour
- Photoperiod (hrs dark / hrs light): Fluorescent light: 12 hours/day
IN-LIFE DATES: From: To: No data
Administration / exposure
- Route of administration:
- oral: feed
- Details on route of administration:
- Because of limited solubility in water, the dye was administered to animals in feed.
- Vehicle:
- other: Feed
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed to give dose levels of 0, 1,000, 3000, 7000, 15000, or 30000 ppm (0, 200, 600, 1400, 3000 or 6000 mg/Kg/day). Dose formulations were prepared prior to the initiation and at the midpoint of the 14-day studies
DIET PREPARATION
- Rate of preparation of diet (frequency): Prior to the initiation and at the midpoint of the 14-day studies
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: 25˚C
VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 0 , 1,000, 3000, 7000, 15000, or 30000 ppm (0, 200, 600, 1400, 3000 or 6000 mg/Kg/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Homogeneity was confirmed and the stability of the dose formulations was established for 3 weeks when stored in the dark at temperatures up to 25° C and for 1 week when stored open to air and light. Periodic analyses of the dose formulations of C.I. Direct Blue 218 were conducted at the study laboratory and at the analytical chemistry laboratory using visible spectroscopy. For the 14-day studies, the dose formulations were analyzed prior to study
initiation - Duration of treatment / exposure:
- 14 days
- Frequency of treatment:
- Daily
Doses / concentrations
- Remarks:
- 0 , 1,000, 3000, 7000, 15000, or 30000 ppm (0, 200, 600, 1400, 3000 or 6000 mg/Kg/day)
- No. of animals per sex per dose:
- Total: 30 males and 30 females
0 mg/Kg/day: 5 males and 5 females
200 mg/Kg/day: 5 males and 5 females
600 mg/Kg/day: 5 males and 5 females
1400 mg/Kg/day: 5 males and 5 females
3000 mg/Kg/day: 5 males and 5 females
6000 mg/Kg/day: 5 males and 5 females - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: No data
- Rationale for animal assignment (if not random): Distributed using a table of random numbers.
- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: No data
- Section schedule rationale (if not random): No data - Positive control:
- No data
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily
- Cage side observations checked in table [No.?] were included. Mortality
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once daily
BODY WEIGHT: Yes
- Time schedule for examinations: at study initiation, weekly, and prior to necropsy
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, once a week
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data
FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data
OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
HAEMATOLOGY: No data
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data
CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data
URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data
NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data
OTHER: No data - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes, The brain, heart, right kidney, liver, lung, right testis, and thymus were weighed.
HISTOPATHOLOGY: Yes, Complete histopathologic examinations were performed on 30,000 ppm (3000 mg/Kg/day) animals. In addition to gross lesions and tissue masses, the tissues examined included: adrenal gland, brain, epididymis, esophagus, gall bladder (mice), large intestine (cecum, colon, rectum), small intestine (duodenum, jejunum, ileum) heart, kidney, liver, lung, mammary gland, mandibular lymph node, mesenteric lymph node, nose, ovary, pancreas, parathyroid gland, pituitary gland, prostate gland, salivary gland, seminal vesicle, skin, spleen, sternebrae, stomach (forestomach and glandular), testes, thyroid gland, trachea, thymus, urinary bladder, and uterus. In addition, the liver, thymus and gallbladder of all mice was also examined. - Other examinations:
- No data
- Statistics:
- No data
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Blue discoloration of the feces was observed in all exposed groups of male and female mice. Male and female mice of the 30000 ppm (6000 mg/Kg/day) appeared emaciated and hyperactive during the last week of the study
- Mortality:
- no mortality observed
- Description (incidence):
- No death was noted during the study period
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Male and female mice exposed to 30,000 ppm (6000 mg/Kg/day) lost weight. The final mean body weights of other exposed groups were similar to those of the controls.
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- Feed consumption by exposed groups was similar to that by controls, except for the last week of the study, when the apparent amount of feed consumed by the 15,000 and 30,000 ppm groups was greater than that consumed by the controls. This increased feed consumption was related to increased feed spillage in these exposure groups.
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Behaviour (functional findings):
- not specified
- Immunological findings:
- not specified
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- In male mice that received 15,000 ppm (3000 mg/Kg/day), the absolute and relative liver weights were significantly greater than those of the controls, and in males exposed to 30,000 ppm (6000 mg/Kg/day) and females exposed to 15,000 and 30,000 ppm (3000 and 6000 mg/Kg/day) the relative liver weights were significantly greater than those of controls. In the 30,000 ppm groups (6000 mg/Kg/day), there were some significantly lower absolute organ weights which were attributed to the lower body weights. The relative heart weight of 30,000 ppm (6000 mg/Kg/day) females and the relative thymus weights of 30,000 ppm (6000 mg/Kg/day) males and females were significantly lower than those of the controls
- Gross pathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- At necropsy, the skin and gastrointestinal tracts of exposed male and female mice had blue discoloration
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- not specified
- Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
- Details on results:
- No data
Effect levels
- Dose descriptor:
- NOAEL
- Effect level:
- 3 000 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No significant toxic effects were noted at the mentioned dose level
Target system / organ toxicity
- Critical effects observed:
- not specified
Any other information on results incl. tables
Table: Survival, Mean Body Weights, and Feed Consumption of Mice in the 14-Day Feed Study of C.I. Direct Blue 218
Dose (ppm) |
Survival |
Mean Body weight |
Final weight relative to controls (%) |
Feed consumption |
|||
Initial |
Final |
Change |
Week 1 |
Week 2 |
|||
Male |
|
|
|
|
|
|
|
0 |
5/5 |
22.3±0.9 |
24.0±0.9 |
17±0.1 |
|
2.4 |
2.6 |
1000 |
5/5 |
22.3±0.7 |
24.5±0.9 |
2.2±0.2 |
102 |
2.8 |
2.8 |
3000 |
5/5 |
22.1±0.7 |
24.2±0.7 |
2.1±0.2 |
101 |
2.6 |
2.5 |
7000 |
5/5 |
22.0±1.0 |
23.9±0.8 |
1.9±0.5* |
100 |
2.6 |
2.9 |
15000 |
5/5 |
22.0±0.9 |
22.3±1.4 |
0.3±0.6** |
93 |
3.4 |
5.0d |
30000 |
5/5 |
22.1±0.9 |
17.9±0.8** |
-4.2±0.3** |
75 |
4.0 |
5.6d |
Females |
|
|
|
|
|
|
|
0 |
5/5 |
17.5±0.4 |
18.8±0.4 |
1.2±0.2 |
|
2.2 |
1.8 |
1000 |
5/5 |
17.4±0.7 |
18.3±0.5 |
0.9±0.3 |
98 |
2.1 |
2.4 |
3000 |
5/5 |
17.8±0.4 |
18.8±0.2 |
1.0±0.3 |
100 |
2.5 |
2.2 |
7000 |
5/5 |
17.6±0.8 |
18.7±0.8 |
1.1±0.2 |
100 |
2.7 |
2.7 |
15000 |
5/5 |
17.1±0.4 |
18.0±0.3 |
0.9±0.4 |
96 |
3.1 |
3.9d |
30000 |
5/5 |
17.9±1.0 |
15.1±0.8** |
-2.8±0.4** |
80 |
3.6 |
4.7d |
* Significantly different (P≤O.05) from the control group by Williams' or Dunnett's test
** P≤0.01
Applicant's summary and conclusion
- Conclusions:
- The No observed adverse effect level (NOAEL) for C.I. Direct blue 218 is considered to be 3000 mg/Kg/day for males and females B6C3F1 mice.
- Executive summary:
14 days repeated dose toxicity study was conducted to determine the toxic nature of C.I. Direct blue 218. The study was performed using male and female B6C3F1 mice and dosing the test chemical in feed at dose levels of0 , 1,000, 3000, 7000, 15000, or 30000 ppm (0, 200, 600, 1400, 3000 or 6000 mg/Kg/day). The animals were observed for clinical signs, body weight, changes in body weight and food consumption and were subjected to gross pathology and histopathology. No animals died during the study period.Blue discoloration of the feces was observed in all exposed groups of male and female mice. Male and female mice of the 30000 ppm (6000 mg/Kg/day) appeared emaciated and hyperactive during the last week of the study. Male and female mice exposed to 30,000 ppm (6000 mg/Kg/day) lost weight. The final mean body weights of other exposed groups were similar to those of the controls. Feed consumption by exposed groups was similar to that by controls, except for the last week of the study, when the apparent amount of feed consumed by the 15,000 and 30,000 ppm groups was greater than that consumed by the controls. This increased feed consumption was related to increased feed spillage in these exposure groups. In male mice that received 15,000 ppm (3000 mg/Kg/day), the absolute and relative liver weights were significantly greater than those of the controls, and in males exposed to 30,000 ppm (6000 mg/Kg/day) and females exposed to 15,000 and 30,000 ppm (3000 and 6000 mg/Kg/day) the relative liver weights were significantly greater than those of controls. In the 30,000 ppm groups (6000 mg/Kg/day), there were some significantly lower absolute organ weights which were attributed to the lower body weights. The relative heart weight of 30,000 ppm (6000 mg/Kg/day) females and the relative thymus weights of 30,000 ppm (6000 mg/Kg/day) males and females were significantly lower than those of the controls. At necropsy, the skin and gastrointestinal tracts of exposed male and female mice had blue discoloration. Based on the observations made, the No observed adverse effect level (NOAEL) for C.I. Direct blue 218 is considered to be 3000 mg/Kg/day for males and females B6C3F1 mice.
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.
![ECHA](/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/echa_logo.png)