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EC number: 221-409-2 | CAS number: 3087-16-9
- 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
Non carcinogenic NOAEL was considered to be 530 and 660 mg/kg body weight/day in males and females when CD-1 male and female mice treated with Green S orally in diet for 2 years.
Key value for chemical safety assessment
Carcinogenicity: via oral route
Link to relevant study records
- Endpoint:
- carcinogenicity: oral
- 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 peer reiviewed journal
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)
- Principles of method if other than guideline:
- Long term chronic and carcinogenicity study of Green S orally in mice.
- GLP compliance:
- not specified
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Green S
- Molecular formula (if other than submission substance): C27H26N2O7S2.Na
- Molecular weight (if other than submission substance): 576.6 g/mole
- Substance type: Organic
- Physical state:
Purity: Dye 82% (80%)
- Impurities (identity and concentrations): 20 %; volatile matter, 3.37% (4.9%); water-insoluble matter, 0.01% (0.07%); sodium chloride, 1.4% (2.6%); sodium
sulphate, 8.5% (6.0%); pH of a 1% solution in distilled water, 5.4 (2.9); lead, <5 ppm; copper, 4ppm (2ppm); chromium, 6ppm (8ppm); zinc, 9ppm (5ppm); iron, 30ppm (35ppm); cadmium, < 1 ppm; mercury, 0.2 ppm (< 1 ppm); free aromatic amines (as aniline), 29ppm (17ppm); Michler's hydrol, <0.01% (<0.02%); Michler's ketone, <0.01%; R-acid, 0.11% (0.05%); subsidiary dyes, < 1%; ether extractable material, 0.14%. - Species:
- mouse
- Strain:
- CD-1
- Details on species / strain selection:
- No data
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- - Source: Specified-pathogen-free colony (Charles River (UK) Ltd, Margate, Kent)
- Age at study initiation: 4-5 week old
- Weight at study initiation: Female : 20.2-20.6 g
- Fasting period before study: No data available
- Housing: Animals were housed in stainless-steel and plastic solid-floored cages with autoclaved softwood sawdust as bedding. The cages were held on mobile racks.
- Diet (e.g. ad libitum): Basal diet (Spratt's Laboratory Diet No. 2-Spratt's Patent Ltd, Barking, Essex)
- Water (e.g. ad libitum): No data available
- Acclimation period: 8 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 3°C
- Humidity (%): 50-70%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available
IN-LIFE DATES: From: To: No data available - Route of administration:
- oral: feed
- Type of inhalation exposure (if applicable):
- not specified
- Vehicle:
- other: Basal diet (Spratt's Laboratory Diet No. 2)
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: Green S was incorporated by dry-blending into the basal diet (Spratt's Laboratory Diet No. 2-Spratt's Patent Ltd, Barking, Essex) to give the required concentration.
DIET PREPARATION
- Rate of preparation of diet (frequency): Once every 6 week
- Mixing appropriate amounts with (Type of food): Basal diet (Spratt's Laboratory Diet No. 2)
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water): Basal diet (Spratt's Laboratory Diet No. 2)
- Concentration in vehicle: 0, 0.033, 0.33 and 0.66%
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- by using a colorimetric estimation of aqueous diet extracts.
- Duration of treatment / exposure:
- 2 year (106 weeks)
- Frequency of treatment:
- Daily
- Post exposure period:
- No data available
- Remarks:
- 0, 49.5, 495 and 530-660 mg/kg bw/day
- No. of animals per sex per dose:
- Total: 510
0 mg/kg bw : 105 male, 105 female
49.5 mg/kg bw: 65 male, 65 female
495 mg/kg bw: 65 male, 65 female
660 mg/kg bw: 65 male
530 mg/kg bw: 65 female - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Rationale for animal assignment (if not random): Animals were assigning to treatment group by using random number chart.
- Positive control:
- No data available
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included: Morbidity and mortality were observed.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily
BODY WEIGHT: Yes
- Time schedule for examinations: On day 1 and weekly thereafter
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available
FOOD EFFICIENCY: No data available
- 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 available
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available
OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
HAEMATOLOGY: Yes
- Time schedule for collection of blood: At 14, 28 and 51 week and at the end of study.
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: 20 animals from each group
- Parameters checked in table [No.?] were examined: Hemoglobin concentration, erythrocyte count, total leucocyte count, packed cell volume, reticulocyte count and differential leucocyte count were examined.
CLINICAL CHEMISTRY: No data available
- Time schedule for collection of blood: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available
URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined. No data available
NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data available
OTHER:
Organ weigth:
Brain, caecum (full and empty), heart, kidneys, liver, spleen, stomach and testes were weighed. - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
Any abnormalities were observed.
Non-neoplastic and neoplastic lesions were observed
Samples of organs were preserved in 10% buffered formalin, along with any other tissue found to be abnormal. One eye was fixed in Davidson's fluid for 24 hr before transfer to 10% buffered formalin.
HISTOPATHOLOGY: Yes
Samples of all tissues taken, except nasal bones from the 0.033 and 0.33% groups, were processed for wax embedding. Sections 5pro thick were prepared, stained with haematoxylin and eosin and examined by light microscopy.
Organ examined:
Brain, caecum (full and empty), heart, kidneys, liver, spleen, stomach, testes, adrenal gland, aorta, bladder, colon, diaphragm, duodenum, ears, epididymis, eye, Harderian gland, ileum, lungs, lymph nodes, mammary gland, skeletal muscle, nasal
bones, nerve, oesophagus, ovaries, pancreas, pituitary gland, prostate, rectum, salivary gland, seminal vesicles, skin, spinal cord, thymus, thyroid, trachea, uterus, vagina and vena cava were examined. - Other examinations:
- Numbers of live and dead pups and gross internal examination were performed.
- Statistics:
- Statistical analysis of Mortality data were analysed according to the method of Peto & Pike (1973). Body weight, organ weight and haematology data were subject to analysis of variance and t test. Litter observations were analysed using Fisher's exact test. Incidences of histopathological findings were compared using the methods of Peto et al. (1980). In all analyses a probability level of less than 5% was taken as statistically significant.
- Clinical signs:
- no effects observed
- Dermal irritation (if dermal study):
- not examined
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- not specified
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- no effects observed
- 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, non-treatment-related
- Gross pathological findings:
- effects observed, treatment-related
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Histopathological findings: neoplastic:
- no effects observed
- Other effects:
- no effects observed
- Details on results:
- Mortality: No effect was observed on survival of all treated mice as compared to control.
Clinical signs: No effect was observed on general condition of all treated mice as compared to control.
Body weight and weight gaiin: No effect was observed on body weight of male mice in all treated groups.
In female mice, significant decrease was observed in body weight on week 9 in 0.66 % dose and on week 87 in 0.33 % dose group, but from this point onwards the difference was reversed and differences were not statistically significant.
Haematology Very few statistically significant differences were observed on 14, 28, 51 and 106 week in treated and control.
The differences present did not occur consistently at all times of examination or in both sexes.
Organ Weights: Increase in absolute stomach weight was observed in female mice at 0.66 % dose but was not present when the weights were expressed relative to body weight.
No other consistent or dose-related statistically significant differences between the treated and control groups.
Gross Pathology: Green coloration of the gastro-intestinal tract was observed in all the treated mice as compared to control.
Histopathology: non-neoplastic Hyperplasia in Caecum, Luminal casts in Epididymis, Focal epithelial hyperplasia of nephron in Kidney, Necrotic/inflammatory foci in Liver and oedema in rectum were observed in male mice at 0.66 % were observed as compared to control.
Pneumonia in Lung at 0.66 % and Hepatitis of Liver in female at 0.33 % was observed as compared to control.
Histopathology: neoplastic Cyst adenoma in Harderian Gland, Interstitial-cell tumors in Testes were observed in male mice as compared to control.
The total numbers of tumours and animals with tumours, calculated for both malignant and benign tumours show no positive dose-related trends.
OTHER FINDINGS: No effects were observed on Numbers of live and dead pups of treated mice as compared to control. - Dose descriptor:
- NOAEL
- Effect level:
- 660 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- male
- Basis for effect level:
- body weight and weight gain
- clinical signs
- food consumption and compound intake
- gross pathology
- haematology
- histopathology: neoplastic
- histopathology: non-neoplastic
- mortality
- organ weights and organ / body weight ratios
- Dose descriptor:
- NOAEL
- Effect level:
- 530 mg/kg bw/day
- Based on:
- test mat.
- Sex:
- female
- Basis for effect level:
- body weight and weight gain
- clinical signs
- food consumption and compound intake
- gross pathology
- haematology
- histopathology: neoplastic
- histopathology: non-neoplastic
- mortality
- organ weights and organ / body weight ratios
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Treatment related:
- not specified
- Dose response relationship:
- not specified
- Relevant for humans:
- not specified
- Conclusions:
- Non carcinogenic NOAEL was considered to be 530 and 660 mg/kg body weight/day in males and females when CD-1 male and female mice treated with Green S orally in diet for 2 years.
- Executive summary:
In a Long term chronic and carcinogenicity study, CD-1 male and female mice treated with Green S in the concentration of 0, 0.033, 0.33 and 0.66 % orally in diet (0, 49.5, 495 and 530 – 660 mg/kg bw). No effects were observed on survival, clinical sign and body weight of treated mice as compared to control. Very few statistically significant differences were observed in hematology on 14, 28, 51 and 106 week, but the differences present did not occur consistently at all times of examination or in both sexes. Increase in absolute stomach weight was observed in female mice at 530 mg/kg bw dose but was not present when the weights were expressed relative to body weight. Similarly, Green coloration of the gastro-intestinal tract was observed in all the treated mice. It is possible that such differences could arise from the study design. Non – neoplastic hyperplasia in Caecum, Luminal casts in Epididymis, Focal epithelial hyperplasia of nephron in Kidney, Necrotic/inflammatory foci in Liver and oedema in rectum and Pneumonia in Lung at 530 mg/kg bw and Hepatitis of Liver at 495 in female mice were observed. In addition, neoplastic cyst adenoma in Harderian Gland, Interstitial-cell tumors in Testes were observed in male mice as compared to control. The total numbers of tumours and animals with tumours, calculated for both malignant and benign tumours show no positive dose-related trends and considered as non carcinogenic. Therefore, NOAEL was considered to be 530 and 660 mg/kg body weight/day in males and females when CD-1 male and female mice treated with Green S orally in diet for 2 years.
Reference
Tumour incidence in mice fed diets containing 0-1100 mg/Kg/day % Gree
|
No. of animals affected in each group |
|||||||
Males |
Females |
|||||||
0 |
55 |
550 |
1100 |
0 |
55 |
550 |
1100 |
|
Total no. of mice |
|
|
|
|
|
|
|
|
Examined |
85 |
50 |
50 |
50 |
85 |
50 |
50 |
50 |
With primary tumours |
56 |
35 |
29 |
38 |
57 |
36 |
35 |
30 |
With malignant tumors |
25 |
10 |
11 |
13 |
232 |
18 |
19 |
16 |
Benign tumors |
45 |
31 |
22 |
30 |
39 |
21 |
20 |
16 |
Total no. of malignant tumours |
25 |
10 |
11 |
13 |
26 |
18 |
20 |
16 |
Total no. of benign tumours |
57 |
51 |
26 |
41 |
56 |
24 |
27 |
18 |
Statistical analysis (Peto et al. 1980) of the number of animals with primary tumours, total number of animals with malignant tumours and total number with benign tumours did not show any positive or negative trends.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 660 mg/kg bw/day
- Study duration:
- chronic
- Species:
- mouse
- Quality of whole database:
- Data is Klimisch 2 and from peer -reivewed journal
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
Based on the above study on Hydrogen [4-[4-(dimethylamino)-α-(2-hydroxy-3,6-disulphonato-1-naphthyl)benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium, monosodium salt, it can be concluded that Hydrogen [4-[4-(dimethylamino)-α-(2-hydroxy-3,6-disulphonato-1-naphthyl)benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium, monosodium salt can be considered as Not classified for carcinogenicity.
Additional information
Carcinogenicity - Oral:
In a study, Hydrogen [4-[4-(dimethylamino)-α-(2-hydroxy-3,6-disulphonato-1-naphthyl)benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium, monosodium salt (Green S) has been investigated for carcinogenicity based on in vivo experiments in rodents, i.e. most commonly in mice for Hydrogen [4-[4-(dimethylamino)-α-(2-hydroxy-3,6-disulphonato-1-naphthyl)benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium, monosodium salt.
In a Long term chronic and carcinogenicity study by Brantomet al(Food Chem. Toxic. Vol. 25, No. 12, pp. 977-983, 1987), CD-1 male and female mice were treated with Green S in the concentration of 0, 49.5, 495 and 530 – 660 mg/kg bw orally in diet. No effects were observed on survival, clinical sign and body weight of treated mice as compared to control. Very few statistically significant differences were observed in hematology on 14, 28, 51 and 106 week, but the differences present did not occur consistently at all times of examination or in both sexes. Increase in absolute stomach weight was observed in female mice at 530 mg/kg bw dose but was not present when the weights were expressed relative to body weight. Similarly, Green coloration of the gastro-intestinal tract was observed in all the treated mice. It is possible that such differences could arise from the study design. Non – neoplastic hyperplasia in Caecum, Luminal casts in Epididymis, Focal epithelial hyperplasia of nephron in Kidney, Necrotic/inflammatory foci in Liver and oedema in rectum and Pneumonia in Lung at 530 mg/kg bw and Hepatitis of Liver at 495 in female mice were observed. In addition, neoplastic cyst adenoma in Harderian Gland, Interstitial-cell tumors in Testes were observed in male mice as compared to control. The total numbers of tumours and animals with tumours, calculated for both malignant and benign tumours show no positive dose-related trends and considered as non carcinogenic. Therefore, non carcinogenic NOAEL was considered to be 530 and 660 mg/kg body weight/day in males and females when CD-1 male and female mice treated with Green S orally in diet for 2 years.
Thus, based on the above study on Hydrogen [4-[4-(dimethylamino)-α-(2-hydroxy-3,6-disulphonato-1-naphthyl)benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium, monosodium salt, it can be concluded that Hydrogen [4-[4-(dimethylamino)-α-(2-hydroxy-3,6-disulphonato-1-naphthyl)benzylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium, monosodium salt can be considered as Not classified for carcinogenicity.
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