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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 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
Carcinogenicity
Administrative data
Description of key information
Exposure to malachite green chloride in feed resulted in nonneoplastic lesions in the thyroid gland and liver of female rats and the urinary bladder of female mice.
Key value for chemical safety assessment
Carcinogenicity: via oral route
Link to relevant study records
- Endpoint:
- carcinogenicity: oral
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- From April 13th, 1999 to May 7th, 2001
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: National Toxicology Program study report. 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.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)
- Principles of method if other than guideline:
- National Toxicology Program study report.
- GLP compliance:
- yes
- Species:
- rat
- Strain:
- Fischer 344
- Sex:
- female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: National Centre for Toxicological Research (NCTR).
- Age at study initiation: approximately 6 weeks old at the beginning of the studies.
- Housing: animals were distributed randomly into groups of approximately equal initial mean body weights. Rats were housed two per cage. Cages were changed once a week and rotated every 3 weeks.
- Cage: polycarbonate cages (Allentown Caging Equipment Co., Allentown, NJ), changed weekly and rotated every 3 weeks.
- Bedding: hardwood chips (Northeastern Products, Inc., Warrensburg, NY),changed weekly.
- Cage Bonnets: microisolator tops.
- Racks: metal animal cage racks, changed weekly.
- Diet: feed was available ad libitum. NIH-31 open formula meal pellets were autoclaved then ground to powder (Purina Mills, Richmond, IN), available ad libitum until the day before sacrifice.
- Water: ad libitum. Millipore-filtered water (Jefferson municipal supply) via 480 ml water bottles.
- Acclimation period: 2 weeks.
- Health check: the health of the animals was monitored during the studies according to the protocols of the Study Laboratory’s Sentinel Animal Program.
ENVIRONMENTAL CONDITIONS
- Temperature: 23 ± 0.5°C
- Humidity: 49.4 %
- Air changes: at least 10/hour
- Photoperiod: 12 hours/day - Route of administration:
- oral: feed
- Vehicle:
- water
- Details on exposure:
- Dose formulations were prepared approximately every 2 months by dissolving the chemical in water and then mixing it with feed. The solution was blended with feed in a Patterson-Kelly V-shell blender using an intensifier bar and a heater under a vacuum of at least 15 mm mercury for approximately 20 minutes.
Dose formulations were stored in stainless steel feed cans at 4 ± 2 °C for up to 92 days. - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- A homogeneity study of a 100 ppm dose formulation and a stability study of a 25 ppm dose formulation were performed by the study laboratory using HPLC. Homogeneity was confirmed, and stability was confirmed for at least 10 days for dose formulations stored at room temperature exposed to light and for at least 92 days for formulations stored at up to 6 °C protected from light.
Periodic analyses of the dose formulations were conducted by the study laboratory using HPLC. The dose formulations were analyzed approximately every 7 weeks. Of the malachite green chloride dose formulations analyzed and used, 96 % (65/68) of the dose formulations were within 10 % of the target concentrations. - Duration of treatment / exposure:
- 104 weeks
- Frequency of treatment:
- Daily
- Remarks:
- Doses / Concentrations:
0, 100, 300, or 600 ppm (equivalent to average daily doses of approximately 0, 7, 21 and 43 mg/kg bw/day)
Basis:
nominal in diet - No. of animals per sex per dose:
- 48 females
- Control animals:
- yes, plain diet
- Details on study design:
- - Dose selection rationale: to assist in selecting doses for the 2-year bioassay, a 28-day range-finding study was conducted in which male and female F344 rats were fed the test item (0, 25, 100, 300, 600, or 1200 ppm) in feed.
- Rationale for animal assignment: animals were distributed randomly into groups of approximately equal initial mean body weights. - Observations and examinations performed and frequency:
- DETAILED CLINICAL OBSERVATIONS
Observed twice daily and clinical findings recorded weekly.
BODY WEIGHT
Animals were weighed initially, weekly for the first 12 weeks, approximately every 4 weeks until week 92, weekly for the last 12 weeks, and at the end of the studies.
FOOD CONSUMPTION AND COMPOUND INTAKE
Feed consumption was recorded weekly for the first 12 weeks and approximately every 4 weeks thereafter. - Sacrifice and pathology:
- Sacrifice: carbon dioxide asphyxiation.
GROSS PATHOLOGY
Necropsies were performed on all animals. Organs weighed were the kidneys and liver and the thyroid gland.
HISTOPATHOLOGY
Complete histopathology was performed on all rats. In addition to gross lesions and tissue masses, the following tissues were examined: adrenal gland, bone marrow (femur, sternum), brain (cerebellum, cerebrum, stem), clitoral gland, esophagus, eye, harderian gland, heart with aorta, large intestine (cecum, colon, rectum), small intestine (duodenum, jejunum, ileum), kidney, liver, lung, lymph nodes (mandibular and mesenteric), mammary gland, muscle (thigh), nerve (sciatic), nose, ovaries, pancreas, parathyroid gland, pituitary gland, salivary gland, skin, spinal cord (thoracic), spleen, stomach (forestomach and glandular), thymus, thyroid gland, tongue, trachea, urinary bladder, uterus, vagina, and Zymbal’s gland. - Statistics:
- Survival Analyses: the probability of survival was estimated by the product-limited procedure of Kaplan and Meier (1958).
Analysis of Neoplasm and Nonneoplastic Lesion Incidences: the Poly-k test (Bailer and Portier, 1988; Portier and Bailer, 1989; Piegorsch and Bailer, 1997) was used to assess neoplasm and nonneoplastic lesion prevalence.
Analysis of Continuous Variables: for both body weights and food consumption, the mixed models approach to repeated measures ANOVA was used. Organ weights, terminal body weights, and the ratios of organ weight to terminal body weight for terminally sacrificed animals were analyzed using ANOVA procedures. Terminal body weights were also used as a covariant in an ANACOVA procedure. For each end point analyzed, Dunnett’s two-sided test (Dunnett, 1955) was used to compare the control group mean to each treatment group mean, either overall or at each point of time, whichever was appropriate. - Clinical signs:
- no effects observed
- Description (incidence and severity):
- similar to the control group
- Mortality:
- no mortality observed
- Description (incidence):
- similar to the control group
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- 300 and 600 ppm groups less than the control group
- Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- similar to the control group
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- relative liver weight increased (600 ppm)
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- thyroid gland: follicle, cyst (0/46, 1/48, 1/47, 3/46); liver: eosinophilic focus (5/48, 10/48, 13/48, 14/48)
- Histopathological findings: neoplastic:
- no effects observed
- Details on results:
- CLINICAL SIGNS AND MORTALITY
Survival of all exposed groups was similar to that of the control group. Survival rates: 29/48, 23/48, 32/48, 25/48.
No clinical findings were attributed to malachite green chloride exposure.
BODY WEIGHT AND WEIGHT GAIN
Mean body weights of female rats exposed to 300 or 600 ppm malachite green chloride were generally less than those of the controls during most of the study.
FOOD CONSUMPTION AND COMPOUND INTAKE
Feed consumption by exposed rats was generally similar to that by controls throughout the study. Dietary concentrations of 100, 300, or 600 ppm resulted in average daily doses of approximately 7, 21, or 43 mg malachite green chloride/kg body weight.
ORGAN WEIGHTS
The relative liver weight was significantly increased in females exposed to 600 ppm malachite green chloride
PATHOLOGY AND STATISTICAL ANALYSES
Thyroid Gland: follicular cell adenomas and carcinomas were observed in female rats exposed to 300 or 600 ppm and the incidences of adenoma or carcinoma (combined) in these groups exceeded the historical control range. A dose-related increasing trend (P=0.049) in the incidence of cystic follicles was observed in exposed rats. Cystic follicles consisted of very large thyroid follicles that were distended with colloid and lined by flattened follicular epithelial cells. Lesions diagnosed as follicular cell hyperplasia were also cystic, but there were small fronds and foci of follicular epithelial cells protruding into distended follicles. Although the increases were not statistically significant, thyroid follicular cell hyperplasia was only observed in rats exposed to malachite green chloride.
Liver: there were modest, but not statistically significant, increases in the incidences of hepatocellular adenoma in female rats exposed to test item. However, the incidences in all groups, including the controls, exceeded the historical control range. Hepatocellular adenomas consisted of well-demarcated lesions that occupied an area greater in size than one hepatic lobule with distinct compression of adjacent parenchyma. A single hepatocellular carcinoma was found in one 300 ppm female and was a large well-demarcated lesion that consisted of anaplastic hepatocytes arranged in a trabecular pattern in some areas. Corresponding increases in the incidences of eosinophilic foci and centrilobular necrosis were observed in female rats exposed. Eosinophilic foci were characterized by distinct, variably sized foci in which hepatocytes were larger than normal due to increased amounts of eosinophilic cytoplasm. The incidence of centrilobular necrosis, in which the necrotic cells were oriented around central veins in the hepatic lobule, exhibited an increasing trend (P=0.002).
Mammary Gland: the incidence of mammary gland carcinoma in female rats exposed to 600 ppm exceeded the historical control range.
Pituitary Gland: there was a statistically significant increase in the incidence of adenoma of the pituitary gland (pars distalis) in female rats exposed to 100 ppm. Incidences of pars distalis adenoma in 100 and 300 ppm females exceeded the historical control range for pars distalis adenoma or carcinoma (combined).
Mononuclear Cell Leukemia: a dose-related decreasing trend in the incidences of mononuclear cell leukemia occurred in female rats exposed, with statistically significant decreased incidences in the 300 and 600 ppm groups. - Relevance of carcinogenic effects / potential:
- Ambiguous
- Dose descriptor:
- NOAEL
- Effect level:
- 7 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- other: incidence of eosinophilic foci in the liver was increased
- Remarks on result:
- other: Effect type: toxicity (migrated information)
- Dose descriptor:
- LOAEL
- Effect level:
- 21 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- other: thyroid follicular cell adenoma and carcinomas, hepatocellular ademonas , mononuclear cell leukemia
- Remarks on result:
- other: Effect type: carcinogenicity (migrated information)
- Conclusions:
- It was concluded that tumors of the thyroid gland, liver, or mammary gland in female rats might have been caused by Malachite Green chloride.
- Executive summary:
Malachite Green chloride effects were studied on female rats to identify potential toxic or cancer-related hazards to humans. The dye was mixed into the feed of rats and the doses given were 100, 300, or 600 ppm (equivalent to average daily doses of approximately 0, 7, 21 and 43 mg/kg bw/day). Control animals received the same feed with no chemical added. The study lasted for two years. Tissues from more than 40 sites were examined for every animal.
Rats exposed to Malachite Green chloride weighed less on average than the control animals. In rats exposed to the dye, there were very slight increases in a few types of tumors: cancers of the thyroid gland, liver, and mammary gland. It was concluded that tumors of the thyroid gland, liver, or mammary gland in female rats might have been caused by Malachite Green chloride.
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- 7 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
Carcinogenicity: via inhalation route
Endpoint conclusion
- Endpoint conclusion:
- no study available
Carcinogenicity: via dermal route
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
According to the CLP Regulation (EC 1272/2008), for the purpose of the classification for Carcinogenicity, section 3.6, substances can be allocated in one of two categories, as follow.
A substance is classified in Category 1, as known or presumed human carcinogens, on the basis of epidemiological and/or animal data. A substance may be classified as known to have carcinogenic potential for humans, classification largely based on human evidence or presumed to have carcinogenic potential for humans, based on animal evidence.
The placing of a substance in Category 2 is done on the basis of evidence obtained from human and/or animal studies, but which is not sufficiently convincing to place the substance in Category 1, based on strength of evidence together with additional considerations.
Based on the available information, Malachite Green has not been judged as presumed either suspected human carcinogen.
In conclusion, the substance does not meet the criteria to be classified for carcinogenicity, according to the CLP Regulation (EC 1272/2008).
Additional information
Under the conditions of the NTP (2005) 2-year feed studies, there was equivocal evidence of carcinogenic activityof malachite green chloride in female F344/N rats based on the occurrence of thyroid gland follicular cell adenoma or carcinoma (combined) and marginal increases in hepatocellular adenoma and mammary gland carcinoma in exposed rats. There wasno evidence of carcinogenic activityof malachite green chloride in female B6C3F1mice exposed to 100, 225, or 450 ppm.
Under the conditions of these 2-year feed studies, there wasequivocal evidence of carcinogenic activityof leucomalachite green in male F344/N rats based on an increase in interstitial cell adenoma of the testes and the occurrence of thyroid gland follicular cell adenoma or carcinoma (combined) in exposed rats. There wasequivocal evidence of carcinogenic activityof leucomalachite green in female F344/N rats based on a marginally increased incidence of hepatocellular adenoma and the occurrence of thyroid gland follicular cell adenoma or carcinoma (combined) in exposed rats. There wassome evidence of carcinogenic activityof leucomalachite green in female B6C3F1mice based on an increase in hepatocellular adenoma or carcinoma (combined) (NTP, 2005).
Exposure to malachite green chloride in feed resulted in nonneoplastic lesions in the thyroid gland and liver of female rats and the urinary bladder of female mice; exposure to leucomalachite green in feed resulted in nonneoplastic lesions in the thyroid gland and liver of male and female rats and the urinary bladder of female mice (NTP, 2005).
Effects attributed to Malachite Green and that seems linked to tumour promotion were reported in literature. The substance appears to be able to produce transformation SHE cells, developing aneuploid pattern; these cells can be tumorigenic, as they could produce tumours (Rao, 2001). Carcinogenesis involves an imbalance between the regulation of cell proliferation and apoptotic death, either by inhibition of apoptosis or by stimulation of cell division, or by affecting both. The Rao et al. study suggests also that transformed cells are much less sensitive to apoptosis than normal cells. Nevertheless, the exact mechanisms by which transformed SHE cells could develop resistance to Malachite Green-induced apoptosis are not clear and further studies are required to see the link between the abrogation of G2/M checkpoint control and the development of resistance to apoptosis observed (Rao, 2001).
Furthermore, static protein tyrosine specific phosphatases associated with enhanced protein tyrosine and/or serine-threonine phosphorylation seems to be linked to abnormal expression of G1/S cyclins and PCNA during rat liver tumour promotion by Malachite Green (Sundarrajan, 2000), but also in this case further studies are required.
Mechanisms behind the induction of these tumours are uncertain and also it is not entirely clear the relevance of these findings.
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.