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EC number: 241-922-5 | CAS number: 18015-76-4
- 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
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in vitro
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2011
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- The experiment is scientifically acceptable, nevertheless the in vitro test followed a not yet validated method (pre-validation on going). Read across from a similar substance which has the same main component and with a different counter ion that does not influence the characteristics related to the specific end-point.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 011
- Report date:
- 2011
Materials and methods
- Principles of method if other than guideline:
- Keratinocytes play a key role in all phases of skin sensitization, and interleukin-18 (IL-18) has been shown to play a key proximal role in the induction of allergic contact sensitization. NCTC 2544 is a commercially available skin epithelial-like cell line originating from normal human skin, which posses a good expression of cytochrome P450-dependent enzymatic activities. THP-1 activation was used to assess the effects on dendritic cells activation (CD68 expression and IL-8 release). Cells were treated with increasing concentrations of both salts (0.25, 0.125, 0.0625, 0.031 µg/ml) or with PPD and LPS as positive controls for 24 h. CD86 expression was assessed by flow cytometry and IL-8 release by ELISA.
- GLP compliance:
- yes
- Type of study:
- other: THP-1 activation and NCTC2544/IL-18 assayes
Test material
- Reference substance name:
- Methanaminium, N-[4-[[4-(dimethylamino)phenyl]phenylmethylene]-2,5-cyclohexadien-1-ylidene]-N-methyl-, ethanedioate
- EC Number:
- 241-922-5
- EC Name:
- Methanaminium, N-[4-[[4-(dimethylamino)phenyl]phenylmethylene]-2,5-cyclohexadien-1-ylidene]-N-methyl-, ethanedioate
- Cas Number:
- 18015-76-4
- Molecular formula:
- C23H25N2 x C2HO4
- IUPAC Name:
- 4-{[4-(dimethylamino)phenyl](phenyl)methylidene}-N,N-dimethylcyclohexa-2,5-dien-1-iminium hydrogen oxalate
- Reference substance name:
- Malachite Green Chloride
- IUPAC Name:
- Malachite Green Chloride
Constituent 1
Constituent 2
Results and discussion
- Positive control results:
- SI: 2.07
IL-8: ca 1600 (pg/ml); CD86: SI ca 2.0
In vitro / in chemico
Results
- Remarks on result:
- other: results are reported in the other sections
Any other information on results incl. tables
Both salts were cytotoxic in both cell lines at concentrations ≤ 1 g/ml.
Results are expressed as mean ± SD of 3-4 independent samples.
According to the prediction model method, a substance is considered as a sensitizer when the SI exceeds the value of 1.2 with a dose-related increase in intracellular IL-18 content. Both salts reach this threshold at concentration between 0.125 - 0.25 µg/ml. In general the dose response of the two salts are similar.
THP-1 ACTIVATION ASSAY
Cells were treated with increasing concentrations of both salts or with LPS as positive control for 24 h. CD86 expression ws assessed by flow cytometry and IL-8 release by ELISA.
Both salts induce a prodes: but statistically significant IL-8 release; furthermore both salts induce CD86 expression (SI≥ 1.5).
NCTC2544/IL-18 ASSAY
The CV80 (0.25 μg/ml) was used as the highest concentration tested for both salts. Intracellular IL-18 was evaluated by ELISA, results were normalized for the cellular protein.
TREATMENT | Intracellular IL-18 (pg/mg) | SI |
Control DMSO | 5256±524 | |
Chl 0.25μg/ml | 6987±634 | 1.33 |
Chl 0.125μg/ml | 6084±758 | 1.16 |
Chl 0.0625μg/ml | 5258±459 | 1.00 |
Chl 0.031μg/ml | 4791±294 | 0.91 |
Oss 0.25μg/ml | 7767±593 | 1.48 |
Oss 0.125μg/ml | 5943±209 | 1.13 |
Oss 0.0625μg/ml | 5700±465 | 1.08 |
Oss 0.031μg/ml | 4925±268 | 0.94 |
PPD 60 μg/ml | 10900±1780 | 2.07 |
Applicant's summary and conclusion
- Interpretation of results:
- sensitising
- Remarks:
- Migrated information Criteria used for interpretation of results: expert judgment
- Conclusions:
- Accordingly to the results, Malachite Green salts should be considered as contact sensitizers.
- Executive summary:
Keratinocytes play a key role in all phases of skin sensitization, and interleukin-18 (IL-18) has been shown to play a key proximal role in the induction of allergic contact sensitization. NCTC 2544 is a commercially available skin epithelial-like cell line originating from normal human skin, which posses a good expression of cytochrome P450-dependent enzymatic activities. THP-1 activation was used to assess the effects on dendritic cells activation (CD68 expression and IL-8 release). Cells were treated with increasing concentrations of both salts (0.25, 0.125, 0.0625, 0.031 µg/ml) or with PPD and LPS as positive controls for 24 h. CD86 expression was assessed by flow cytometry and IL-8 release by ELISA.
According to the prediction model method, a substance is considered as a sensitizer when the SI exceeds the value of 1.2 with a dose-related increase in intracellular IL-18 content. Both salts reach this threshold at concentration between 0.125 - 0.25 µg/ml. In general the dose response of the two salts are similar. Furthermore, statistically significant release fo IL-8 vs vehicle treated cells and stimulation index ≥ 1.5 for CD86 expression are considered as positive results; both salts induce a prodes, but statistically significant IL-8 release and both salts induce CD86 expression (SI > 1.5), thuis they should be classified as senstitizers.
Conclusion
Malachite Green salts should be considered as contact sensitizers.
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