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EC number: 272-939-6 | CAS number: 68921-42-6
- 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
Endpoint summary
Administrative data
Description of key information
Repeated dose toxicity: Oral
Based on the data available, the No Observed Adverse Effect Level (NOAEL) for the test chemical was considered to be in range of 500 -2500 mg/Kg bw using male and female rats or mice . Hence the test chemical is not likely to be toxic upon repeated exposure by oral route as per the criteria mentioned in CLP regulation.
Repeated dose toxicity: inhalation
The melting point of test chemical is >300 °C.This suggests that test chemical decomposes above300 °C without melting and also it does not exhibit very high vapour pressure. Inhalation is therefore not the likely route of exposure and hence this end point is considered for waiver.
Repeated dose toxicity: dermal
Repeated dermal toxicity is unlikely to occur since dermal absorption of test chemical is limited. Also considering the use of the chemical as a colouring agent in food and cosmetics and considering the volatility absorption by the dermal route is not considered to be significant. Thus, given the above considerations, it is assumed that test chemical shall not exhibit acute dose toxicity by the repeated dermal route.
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- chronic toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Remarks:
- Experimental data from various test chemicals
- Justification for type of information:
- Weight of evidence approach based on the available information from various test chemicals.
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- WoE report is based on repeated dose toxicity studies on rats
- GLP compliance:
- not specified
- Limit test:
- no
- Species:
- other: 2.rat 3.mouse
- Strain:
- other: 2.Osborne-Mendel 3.ASH-CS1 strain
- Details on species / strain selection:
- No data
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- 2.TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: Animals were housed individually
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: No data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%): No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
3.TEST ANIMALS
- Source: SPF colony
- Age at study initiation: No data
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: The animals were housed in cages with solid floors. The female mice were caged in groups of five and the male mice individually
- Diet (e.g. ad libitum): reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: No data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 1 deg C
- Humidity (%): 50-60%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
IN-LIFE DATES: From: To: No data - Route of administration:
- oral: feed
- Details on route of administration:
- No data
- Vehicle:
- other: Feed
- Details on oral exposure:
- 2.PREPARATION OF DOSING SOLUTIONS: The test compound is mixed with feed at dose levels of 0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day)
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data
VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data
3.mg/Kg/day)
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): Reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite
- Storage temperature of food: No data
VEHICLE
- Justification for use and choice of vehicle (if other than water): Reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite
- Concentration in vehicle: 0, 70, 700 or 3500 ppm (0, 10, 100 or 500 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:
- not specified
- Details on analytical verification of doses or concentrations:
- No data
- Duration of treatment / exposure:
- 2.
2 years
3.80 weeks - Frequency of treatment:
- Daily
- Remarks:
- Study 2.
0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day)
Study 3.
0, 70, 700 or 3500 ppm (0, 10, 100 or 500 mg/Kg/day) - No. of animals per sex per dose:
- 2.Total -240 male and females
0.0%(0 mg/kg bw/day) -24 male & 24 Female
0.5 %(250 mg/kg bw/day) -24 male & 24Female
1.0%(500 mg/kg bw/day) -24 male & 24 Female
2.0 %(1000 mg/kg bw/day) -24 male & 24 Female
5.0%(2500 mg/kg bw/day) -24 male & 24 Female
3.0 mg/Kg/day: 48 males and 50 females
10 mg/Kg/day: 48 males and 50 females
100 mg/Kg/day: 48 males and 50 females
500 mg/Kg/day: 48 males and 50 females - Control animals:
- yes, concurrent vehicle
- Details on study design:
- No data
- Positive control:
- No data
- Observations and examinations performed and frequency:
- 2.CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the 2 yr study
- Cage side observations checked in table [No.?] were included. Mortality and abnormality
DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data
BODY WEIGHT: Yes
- Time schedule for examinations: Weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- 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
3.CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the study period
- Cage side observations checked in table [No.?] were included. mortality
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data
BODY WEIGHT: Yes
- Time schedule for examinations: At intervals throughout the experiment
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- 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: Yes
- Time schedule for examinations: once during the F0 generation, at the start of the lifetime study, and after 3, 6, 12, 18 and 24 months of the chronic phase
- Dose groups that were examined: All rats
OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 12, 28, 52 wk and 80 wk
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10 animals from 100 and 500 mg/Kg/day
- Parameters checked in table [No.?] were examined. Counts were made of erythrocytes, reticulocytes and total and differential leucocytes, and any abnormalities in either red or white cell morphology, hemoglobin and packed cell volume noted. At 80 wk, blood samples taken from the
caudal vein of all surviving mice were examined for haemoglobin and differential leucocyte counts only.
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
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: Yes
- Time schedule for collection of blood: 3-, 11-, 17-, and 22-month intervals.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10 animals
- Parameters checked in table [No.?] were examined. white blood cell count, hemoglobin, hematocrit, and differential cell count
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:
- 2.GROSS PATHOLOGY: Yes, Survivors were autopsied and organ weights were recorded for heart, liver, spleen, kidney, and testes. Animals that died during the experiment were autopsied, but their organ weights were not considered in the organ weight calculations.
3.GROSS PATHOLOGY: Yes, autopsy was conducted on all animals dying or killed during the experiment unless this was precluded by advanced autolysis or cannibalism. The surviving animals were killed by exsanguination from the aorta under barbiturate anaesthesia after 80 wk and all organs were examined for gross abnormalities. The weights of brain, heart, liver, spleen, kidneys, stomach and small intestine were recorded and samples of these organs, together with samples of salivary gland, thyroid, thymus, adrenals, lymph nodes, pancreas, pituitary, ovaries, uterus, urinary bladder, lungs, testes, duodenum, ileum, colon, caecum, rectum,
spinal cord, skeletal muscle and any other tissue that appeared to be abnormal at autopsy, were preserved in 10% buffered formalin.
HISTOPATHOLOGY: Yes, Paraffin-wax sections of brain, heart, liver, spleen, kidneys, stomach and small intestine, salivary gland, thyroid, thymus, adrenals, lymph nodes, pancreas, pituitary, ovaries, uterus, urinary bladder, lungs, testes, duodenum, ileum, colon, caecum, rectum, spinal cord, skeletal muscle and any other tissue that appeared to be abnormal at autopsy were stained with haematoxylin and eosin and examined for any histopathological changes.
At the end of the two-year feeding study, 225 of the 240 animals started on the experiment were examined for gross pathology.
HISTOPATHOLOGY: Yes, At the end of the two-year feeding study, 225 of the 240 animals started on the experiment were examined for microscopic pathology.
Detailed microscopic examination of lung, heart, stomach, liver, spleen small intestine, colon, bone marrow,leg bone and muscle, urinary bladder, kidney, pancreas, thyroid ( (parathyroid when present), adrenal, and prostate and testis or ovary and uterus was conducted on the formalin-fixed, haematoxylin - eosin stained paraffin embedded tissues of 21 rats from the Blue no. 1. A more limited group of tissues (kidney, liver, testis, and spleen) was examined from 43 rats. - Other examinations:
- No data
- Statistics:
- No data
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- 3.The ingestion of test chemical produced no adverse effects on the behaviour of the animals over 80 wk. The faeces of the treated mice were violet-coloured, the intensity of this colouring being proportional to the dose, but the urine did not appear to be coloured. The fur of the treated mice at the highest dietary level was coloured as a result of contamination by the diet and faeces.
- Mortality:
- no mortality observed
- Description (incidence):
- 2.No effect was observed on mortality of the animals at dietary levels up to 5% (2500 mg/Kg bw/day).
3.No treatment related mortality was noted - Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- 2.No significant growth inhibition occurred with the animals fed test chemical
3.No treatment related body weight changes were noted - 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:
- effects observed, treatment-related
- Description (incidence and severity):
- 2.No effect was observed on of animals at dietary levels up to 5% (2500 mg/Kg bw/day).
3.3.There were significant increases in total white cells in female mice receiving test chemical at dietary levels of 700 or 3500 ppm at wk 12. This appeared to be an isolated effect and could not be demonstrated after 28 or 52 wk. However, at wk 52 there was a statistically significant increase in total white cells and erythrocytes in male mice fed Violet 6B at 3500 ppm of the diet and in the packed cell volume of the females fed test chemical at 700 ppm of the diet. - 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:
- no effects observed
- Description (incidence and severity):
- 2.No effects was observed on weights of heart, liver, kidney, spleen, and testis of the animals that received Blue No. 1 at dietary levels up to 5% (2500 mg/Kg bw/day).
3.The only statistically significant changes of organ weight were in the small intestine of the male mice. Those given diets containing 100 or 500 mg/Kg/day were lower, by 10 and 12% respectively, than the controls. This effect was not statistically significant when the weights were expressed relative to body weight. - Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 2.There was no treatment-related change in gross pathology. The usual gross pathologic conditions of old rats were seen in all groups, with no significant difference in distribution. The most common lesions were tumors, pneumonia (mostly chronic) of varying degrees? chronic nephritis of the usual type seen in our older rats, and testicular atrophy.
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- 2.There was no treatment-related change in microscopical examination. The most common tumors were mammary adenocarcinomas, fibroadenomas, and pulmonary lymphosarcomas.
In addition to the above mentioned common tumors, the Blue No. 1 animals showed the following tumors: at the 2500 mg/Kg bw/day level, one animal each had a hepatoma, a papillary colloid adenoma of the thyroid, and a carcinoma of the prostate; one animal listed with a mammary adenocarcinoma also had a subcutaneous fibroma. At the 1000 mg/kg bw/day level, one animal each had an adenocarcinoma of the kidney, a liposarcoma of the posterior mediastinum, and an extensive lymphoblastoma of the abdominal serosa and lymph nodes. At the 500 mg/kg bw/day level, one animal each had an adrenal cortical adenoma and a subcutaneous malignant angioma. At the 250 mg/Kg bw/day level, one animal each had a subcutaneous lipoma and a mammary carcinoma. In the control group, one animal had a cecal adenoma. The common incidental lesions were pneumonia (mostly chronic) of varying degrees and chronic nephritis of the usual type seen in our older rats There was a moderate degree of testicular atrophy in one of the rats in the 1000 mg/Kg bw/day group, and there was marked atrophy in one of the controls; remaining sections of testis showed no more than slight atrophy. Other major incidental lesions included an ulcerated inflammatory mass in the cecum of a rat on the 2500 mg/Kg bw/day diet, and three large, chronic, tubo-ovarian abscesses( one each on the 1000 and 250 mg/Kg bw/day diets, and control groups)
3.Most of the lesions were those normally occurring in ageing mice and were found with a similar incidence in treated and control mice. None of the findings appeared to be related to the treatment with test chemical . The changes found in the skin consisted of hyperkeratosis, also present in control animals, and a single dermoid cyst in a female given 700 ppm test chemical . Dilated gastric glands and hyperplasia of the gastric mucosa were encountered, but again similar lesions were found in control mice.
Pulmonary adenomas were seen in 36% of all male mice and in 20% of all female mice, the incidence being similar in all groups. Two thyroid adenomas were found in male mice fed Violet 6B (one at 10 and the other at 500 mg/Kg/day). Benign tumours of the ovary, testis, subcutaneous tissue and kidney were found with a similar incidence in control and treated animals. Pulmonary adenocarcinomas were present in two male mice fed diets containing Violet 6B (one at 100 and one at 500 mg/Kg/day). Lymphosarcoma and reticulum-cell neoplasia had a similar incidence in mice fed Violet 6B and in the controls. Three squamous-cell carcinomas
of the skin were seen in female mice (one control, one fed 10 mg/Kg/day and one fed 100 mg/Kg/day Violet 6B). The other malignant tumours found were a fibrosarcoma in a control female, a rhabdomyosarcoma in a female given 10 mg/Kg/day Violet 6B and a mammary adenocarcinoma in a female given 100 mg/Kg/day Violet 6B. - Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
- Details on results:
- No data
- Dose descriptor:
- NOAEL
- Effect level:
- > 500 - <= 2 500 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No significant tretament related alterations were noted at the mentioned dose level
- Remarks on result:
- other: No toxic effects were observed
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Conclusions:
- The No observed Adverse Effect level (NOAEL) for test chemical was considered to be in range of 500-2500 mg/kg bw/day in male and female rats or mice for 2 years feeding study.
- Executive summary:
Data available for the test chemical was reviewed to determine the toxic nature of test chemical .The studies are as mentioned below:
Study 2.
Chronic repeated dose oral toxicity study was performed for test chemical to determine its toxic nature upon repeated exposure. The test chemical was mixed with feed at dose levels of0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day) and fed for 2 years to 24 male and 24 female/dose Osborne Mendel weanling rats. During the study period, the animals were observed for mortality, any abnormality, body weight changes, hematology and were subjected to gross and microscopic pathology. No mortality and abnormality was noted during the study period. No significant growth inhibition occurred with the animals fed with test chemical . Also, no effects was observed on hemotological parameters and weights of heart, liver, kidney, spleen, and testis of the animals that received test chemical at dietary levels up to 5%(2500 mg/Kg bw/day). There was no treatment-related change in gross pathology. The usual gross pathologic conditions of old rats were seen in all groups, with no significant difference in distribution. The most common lesions were tumors, pneumonia (mostly chronic) of varying degrees? chronic nephritis of the usual type seen in our older rats, and testicular atrophy. There was no treatment-related change in microscopical examination. The most common tumors were mammary adenocarcinomas, fibroadenomas, and pulmonary lymphosarcomas. Based on the above considerations, the No observed Adverse Effect level (NOAEL) for test chemical was considered to be 2500 mg/kg bw/day in Osborne-Mendel male and female rats for 2 years feeding study.
Study 3.
Chronic repeated dose toxicity study was performed to determine the oral toxic nature of test chemical using ASH-CSI male and female mice. The test chemical was mixed with appropriate quantity of corn oil and mixed with feed at dose levels of 0, 10, 100 or 500 mg/kg/day. The animals were the test chemical in diet for 80 weeks and during the study the animals were observed for clinical signs, mortality, changes in body weight, hematology and the animals were subjected to gross and histopathology. The faeces of all treated mice were violet-coloured but the urine appeared normal, suggesting that the dose was largely unabsorbed from the gastro-intestinal tract. The feeding of Violet 6B did not affect the mortality, the rate of body-weight gain or the haematological picture at 12, 28, 52 or 80 wk. The only statistically significant changes of organ weight were in the small intestine of the male mice. Those given diets containing 100 or 500 mg/Kg/day were lower, by 10 and 12% respectively, than the controls. This effect was not statistically significant when the weights were expressed relative to body weight. Most of the lesions were those normally occurring in ageing mice and were found with a similar incidence in treated and control mice and appeared to be not related to the treatment with test chemical . The changes found in the skin consisted of hyperkeratosis, also present in control animals, and a single dermoid cyst in a female given 700 ppm test chemical Dilated gastric glands and hyperplasia of the gastric mucosa were encountered, but again similar lesions were found in control mice. Pulmonary adenomas were seen in 36% of all male mice and in 20% of all female mice, the incidence being similar in all groups. Two thyroid adenomas were found in male mice fed test chemical (one at 10 and the other at 500 mg/Kg/day). Benign tumours of the ovary, testis, subcutaneous tissue and kidney were found with a similar incidence in control and treated animals. Pulmonary adenocarcinomas were present in two male mice fed diets containing test chemical (one at 100 and one at 500 mg/Kg/day). Lymphosarcoma and reticulum-cell neoplasia had a similar incidence in mice fed test chemical and in the controls. Three squamous-cell carcinomas of the skin were seen in female mice (one control, one fed 10 mg/Kg/day and one fed 100 mg/Kg/day ). The other malignant tumours found were a fibrosarcoma in a control female, a rhabdomyosarcoma in a female given 10 mg/Kg/day test chemical and a mammary adenocarcinoma in a female given 100 mg/Kg/day of test chemical . Since no deviation was noted in the treated animals from the controls used, hence, the the no observed adverse effect level (NOAEL) for test chemical was considered to be 500 mg/Kg/day.
- Endpoint:
- chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- 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 peer reviwed publication
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- Chronic repeated dose oral toxicity study was performed for test chemical to determine its toxic nature upon repeated exposure
- GLP compliance:
- not specified
- Limit test:
- no
- Specific details on test material used for the study:
- - Name of test material: Brilliant Blue FCF/ Blue no. 1
- Molecular formula: C37H36N2O9S3.2Na
- Molecular weight: 792.8586 g/mol
- Substance type: Organic
- Physical state: No data
- Impurities (identity and concentrations): No data - Species:
- rat
- Strain:
- Osborne-Mendel
- Details on species / strain selection:
- No data
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: Animals were housed individually
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: No data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%): No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
IN-LIFE DATES: From: To: No data - Route of administration:
- oral: feed
- Details on route of administration:
- No data
- Vehicle:
- other: Feed
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test compound is mixed with feed at dose levels of 0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day)
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data
VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data - Analytical verification of doses or concentrations:
- not specified
- Details on analytical verification of doses or concentrations:
- No data
- Duration of treatment / exposure:
- 2 years
- Frequency of treatment:
- Daily
- Remarks:
- 0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day)
- No. of animals per sex per dose:
- Total -240 male and females
0.0%(0 mg/kg bw/day) -24 male & 24 Female
0.5 %(250 mg/kg bw/day) -24 male & 24Female
1.0%(500 mg/kg bw/day) -24 male & 24 Female
2.0 %(1000 mg/kg bw/day) -24 male & 24 Female
5.0%(2500 mg/kg bw/day) -24 male & 24 Female - Control animals:
- yes, concurrent vehicle
- Details on study design:
- No data
- Positive control:
- No data
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the 2 yr study
- Cage side observations checked in table [No.?] were included. Mortality and abnormality
DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data
BODY WEIGHT: Yes
- Time schedule for examinations: Weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- 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: Yes
- Time schedule for collection of blood: 3-, 11-, 17-, and 22-month intervals.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10 animals
- Parameters checked in table [No.?] were examined. white blood cell count, hemoglobin, hematocrit, and differential cell count
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, Survivors were autopsied and organ weights were recorded for heart, liver, spleen, kidney, and testes. Animals that died during the experiment were autopsied, but their organ weights were not considered in the organ weight calculations.
At the end of the two-year feeding study, 225 of the 240 animals started on the experiment were examined for gross pathology.
HISTOPATHOLOGY: Yes, At the end of the two-year feeding study, 225 of the 240 animals started on the experiment were examined for microscopic pathology.
Detailed microscopic examination of lung, heart, stomach, liver, spleen small intestine, colon, bone marrow,leg bone and muscle, urinary bladder, kidney, pancreas, thyroid ( (parathyroid when present), adrenal, and prostate and testis or ovary and uterus was conducted on the formalin-fixed, haematoxylin - eosin stained paraffin embedded tissues of 21 rats from the Blue no. 1. A more limited group of tissues (kidney, liver, testis, and spleen) was examined from 43 rats. - Other examinations:
- No data
- Statistics:
- No data
- Clinical signs:
- not specified
- Mortality:
- no mortality observed
- Description (incidence):
- No effect was observed on mortality of the animals at dietary levels up to 5% (2500 mg/Kg bw/day).
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- No significant growth inhibition occurred with the animals fed test chemical
- 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
- Description (incidence and severity):
- No effect was observed on of animals at dietary levels up to 5% (2500 mg/Kg bw/day).
- 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:
- no effects observed
- Description (incidence and severity):
- No effects was observed on weights of heart, liver, kidney, spleen, and testis of the animals that received Blue No. 1 at dietary levels up to 5% (2500 mg/Kg bw/day).
- Gross pathological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There was no treatment-related change in gross pathology. The usual gross pathologic conditions of old rats were seen in all groups, with no significant difference in distribution. The most common lesions were tumors, pneumonia (mostly chronic) of varying degrees? chronic nephritis of the usual type seen in our older rats, and testicular atrophy.
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There was no treatment-related change in microscopical examination. The most common tumors were mammary adenocarcinomas, fibroadenomas, and pulmonary lymphosarcomas.
In addition to the above mentioned common tumors, the Blue No. 1 animals showed the following tumors: at the 2500 mg/Kg bw/day level, one animal each had a hepatoma, a papillary colloid adenoma of the thyroid, and a carcinoma of the prostate; one animal listed with a mammary adenocarcinoma also had a subcutaneous fibroma. At the 1000 mg/kg bw/day level, one animal each had an adenocarcinoma of the kidney, a liposarcoma of the posterior mediastinum, and an extensive lymphoblastoma of the abdominal serosa and lymph nodes. At the 500 mg/kg bw/day level, one animal each had an adrenal cortical adenoma and a subcutaneous malignant angioma. At the 250 mg/Kg bw/day level, one animal each had a subcutaneous lipoma and a mammary carcinoma. In the control group, one animal had a cecal adenoma. The common incidental lesions were pneumonia (mostly chronic) of varying degrees and chronic nephritis of the usual type seen in our older rats There was a moderate degree of testicular atrophy in one of the rats in the 1000 mg/Kg bw/day group, and there was marked atrophy in one of the controls; remaining sections of testis showed no more than slight atrophy. Other major incidental lesions included an ulcerated inflammatory mass in the cecum of a rat on the 2500 mg/Kg bw/day diet, and three large, chronic, tubo-ovarian abscesses( one each on the 1000 and 250 mg/Kg bw/day diets, and control groups) - Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
- Details on results:
- No data
- Dose descriptor:
- NOAEL
- Effect level:
- 2 500 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No significant tretament related alterations were noted at the mentioned dose level
- Remarks on result:
- other: No toxic effects were observed
- Critical effects observed:
- no
- Conclusions:
- Based on all the available data, it was concluded that the No observed Adverse Effect level (NOAEL) for test chemical was considered to be 2500 mg/kg bw/day in Osborne-Mendel male and female rats for 2 years feeding study.
- Executive summary:
Chronic repeated dose oral toxicity study was performed for test chemical to determine its toxic nature upon repeated exposure. The test chemical was mixed with feed at dose levels of0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day) and fed for 2 years to 24 male and 24 female/dose Osborne Mendel weanling rats. During the study period, the animals were observed for mortality, any abnormality, body weight changes, hematology and were subjected to gross and microscopic pathology. No mortality and abnormality was noted during the study period. No significant growth inhibition occurred with the animals fed with test chemical . Also, no effects was observed on hemotological parameters and weights of heart, liver, kidney, spleen, and testis of the animals that received test chemical at dietary levels up to 5%(2500 mg/Kg bw/day). There was no treatment-related change in gross pathology. The usual gross pathologic conditions of old rats were seen in all groups, with no significant difference in distribution. The most common lesions were tumors, pneumonia (mostly chronic) of varying degrees? chronic nephritis of the usual type seen in our older rats, and testicular atrophy. There was no treatment-related change in microscopical examination. The most common tumors were mammary adenocarcinomas, fibroadenomas, and pulmonary lymphosarcomas. Based on the above considerations, the No observed Adverse Effect level (NOAEL) for test chemical was considered to be 2500 mg/kg bw/day in Osborne-Mendel male and female rats for 2 years feeding study.
- Endpoint:
- chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- 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 peer reviewed publication
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 452 (Chronic Toxicity Studies)
- Principles of method if other than guideline:
- Equivalent or similar to OECD Guideline 452 (Chronic Toxicity Studies)
- GLP compliance:
- not specified
- Limit test:
- no
- Specific details on test material used for the study:
- - Name of test material : Violet 6B (FD & C Violet No. 1)
- Molecular formula : C39H41N3O6S2.Na
- Molecular weight : 734.89 g/mol
- Substance type: Organic
- Physical state: No data
- Impurities: pure dye (by titration with titanium trichloride), min. 85~o; matter volatile at 135°C, max 8%; matter insoluble in water, max 0.3%; ether-extractable matter, max 0.4%; p-dimethyl aminobenzoic acid, max 0.2%; chloride and sulphates of sodium, max 4.0% mixed oxides, max 1.0%; the sum of volatile matter, chloride and sulphates of sodium, pure dye and leuco base of the dye, rain. 95%. - Species:
- mouse
- Strain:
- other: ASH-CS1 strain
- Details on species / strain selection:
- No data
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: SPF colony
- Age at study initiation: No data
- Weight at study initiation: No data
- Fasting period before study: No data
- Housing: The animals were housed in cages with solid floors. The female mice were caged in groups of five and the male mice individually
- Diet (e.g. ad libitum): reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: No data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 1 deg C
- Humidity (%): 50-60%
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
IN-LIFE DATES: From: To: No data - Route of administration:
- oral: feed
- Details on route of administration:
- No data
- Vehicle:
- other: Feed (Reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite)
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed at dose levels of 0, 70, 700 or 3500 ppm (0, 10, 100 or 500 mg/Kg/day)
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): Reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite
- Storage temperature of food: No data
VEHICLE
- Justification for use and choice of vehicle (if other than water): Reground Oxoid pasteurized breeding diet supplemented with 40 ppm sodium menadione bisulphite
- Concentration in vehicle: 0, 70, 700 or 3500 ppm (0, 10, 100 or 500 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:
- not specified
- Details on analytical verification of doses or concentrations:
- No data
- Duration of treatment / exposure:
- 80 weeks
- Frequency of treatment:
- Daily
- Remarks:
- 0, 70, 700 or 3500 ppm (0, 10, 100 or 500 mg/Kg/day)
- No. of animals per sex per dose:
- 0 mg/Kg/day: 48 males and 50 females
10 mg/Kg/day: 48 males and 50 females
100 mg/Kg/day: 48 males and 50 females
500 mg/Kg/day: 48 males and 50 females - Control animals:
- yes, concurrent vehicle
- Details on study design:
- No data
- Positive control:
- No data
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: During the study period
- Cage side observations checked in table [No.?] were included. mortality
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data
BODY WEIGHT: Yes
- Time schedule for examinations: At intervals throughout the experiment
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- 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: Yes
- Time schedule for examinations: once during the F0 generation, at the start of the lifetime study, and after 3, 6, 12, 18 and 24 months of the chronic phase
- Dose groups that were examined: All rats
OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 12, 28, 52 wk and 80 wk
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10 animals from 100 and 500 mg/Kg/day
- Parameters checked in table [No.?] were examined. Counts were made of erythrocytes, reticulocytes and total and differential leucocytes, and any abnormalities in either red or white cell morphology, hemoglobin and packed cell volume noted. At 80 wk, blood samples taken from the
caudal vein of all surviving mice were examined for haemoglobin and differential leucocyte counts only.
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, autopsy was conducted on all animals dying or killed during the experiment unless this was precluded by advanced autolysis or cannibalism. The surviving animals were killed by exsanguination from the aorta under barbiturate anaesthesia after 80 wk and all organs were examined for gross abnormalities. The weights of brain, heart, liver, spleen, kidneys, stomach and small intestine were recorded and samples of these organs, together with samples of salivary gland, thyroid, thymus, adrenals, lymph nodes, pancreas, pituitary, ovaries, uterus, urinary bladder, lungs, testes, duodenum, ileum, colon, caecum, rectum,
spinal cord, skeletal muscle and any other tissue that appeared to be abnormal at autopsy, were preserved in 10% buffered formalin.
HISTOPATHOLOGY: Yes, Paraffin-wax sections of brain, heart, liver, spleen, kidneys, stomach and small intestine, salivary gland, thyroid, thymus, adrenals, lymph nodes, pancreas, pituitary, ovaries, uterus, urinary bladder, lungs, testes, duodenum, ileum, colon, caecum, rectum, spinal cord, skeletal muscle and any other tissue that appeared to be abnormal at autopsy were stained with haematoxylin and eosin and examined for any histopathological changes. - Other examinations:
- No data
- Statistics:
- No data
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- The ingestion of test chemical produced no adverse effects on the behaviour of the animals over 80 wk. The faeces of the treated mice were violet-coloured, the intensity of this colouring being proportional to the dose, but the urine did not appear to be coloured. The fur of the treated mice at the highest dietary level was coloured as a result of contamination by the diet and faeces.
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- No treatment related mortality was noted
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- No treatment related body weight changes were noted
- 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:
- effects observed, treatment-related
- Description (incidence and severity):
- There were significant increases in total white cells in female mice receiving test chemical at dietary levels of 700 or 3500 ppm at wk 12. This appeared to be an isolated effect and could not be demonstrated after 28 or 52 wk. However, at wk 52 there was a statistically significant increase in total white cells and erythrocytes in male mice fed Violet 6B at 3500 ppm of the diet and in the packed cell volume of the females fed test chemical at 700 ppm of the diet.
- 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:
- no effects observed
- Description (incidence and severity):
- The only statistically significant changes of organ weight were in the small intestine of the male mice. Those given diets containing 100 or 500 mg/Kg/day were lower, by 10 and 12% respectively, than the controls. This effect was not statistically significant when the weights were expressed relative to body weight.
- Gross pathological findings:
- not specified
- Neuropathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- not specified
- Histopathological findings: neoplastic:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Most of the lesions were those normally occurring in ageing mice and were found with a similar incidence in treated and control mice. None of the findings appeared to be related to the treatment with test chemical . The changes found in the skin consisted of hyperkeratosis, also present in control animals, and a single dermoid cyst in a female given 700 ppm test chemical . Dilated gastric glands and hyperplasia of the gastric mucosa were encountered, but again similar lesions were found in control mice.
Pulmonary adenomas were seen in 36% of all male mice and in 20% of all female mice, the incidence being similar in all groups. Two thyroid adenomas were found in male mice fed Violet 6B (one at 10 and the other at 500 mg/Kg/day). Benign tumours of the ovary, testis, subcutaneous tissue and kidney were found with a similar incidence in control and treated animals. Pulmonary adenocarcinomas were present in two male mice fed diets containing Violet 6B (one at 100 and one at 500 mg/Kg/day). Lymphosarcoma and reticulum-cell neoplasia had a similar incidence in mice fed Violet 6B and in the controls. Three squamous-cell carcinomas
of the skin were seen in female mice (one control, one fed 10 mg/Kg/day and one fed 100 mg/Kg/day Violet 6B). The other malignant tumours found were a fibrosarcoma in a control female, a rhabdomyosarcoma in a female given 10 mg/Kg/day Violet 6B and a mammary adenocarcinoma in a female given 100 mg/Kg/day Violet 6B. - Other effects:
- not specified
- Dose descriptor:
- NOAEL
- Effect level:
- 500 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No significant treatment related effects were noted at the mentioned dose level
- Remarks on result:
- other: No toxic effects were observed
- Critical effects observed:
- no
- Conclusions:
- Based on all the available data, it was concluded that the no observed adverse effect level (NOAEL) for test chemical was considered to be 500 mg/kg bw/day.
- Executive summary:
Chronic repeated dose toxicity study was performed to determine the oral toxic nature of test chemical using ASH-CSI male and female mice. The test chemical was mixed with appropriate quantity of corn oil and mixed with feed at dose levels of 0, 10, 100 or 500 mg/kg/day. The animals were the test chemical in diet for 80 weeks and during the study the animals were observed for clinical signs, mortality, changes in body weight, hematology and the animals were subjected to gross and histopathology. The faeces of all treated mice were violet-coloured but the urine appeared normal, suggesting that the dose was largely unabsorbed from the gastro-intestinal tract. The feeding of Violet 6B did not affect the mortality, the rate of body-weight gain or the haematological picture at 12, 28, 52 or 80 wk. The only statistically significant changes of organ weight were in the small intestine of the male mice. Those given diets containing 100 or 500 mg/Kg/day were lower, by 10 and 12% respectively, than the controls. This effect was not statistically significant when the weights were expressed relative to body weight. Most of the lesions were those normally occurring in ageing mice and were found with a similar incidence in treated and control mice and appeared to be not related to the treatment with test chemical . The changes found in the skin consisted of hyperkeratosis, also present in control animals, and a single dermoid cyst in a female given 700 ppm test chemical Dilated gastric glands and hyperplasia of the gastric mucosa were encountered, but again similar lesions were found in control mice. Pulmonary adenomas were seen in 36% of all male mice and in 20% of all female mice, the incidence being similar in all groups. Two thyroid adenomas were found in male mice fed test chemical (one at 10 and the other at 500 mg/Kg/day). Benign tumours of the ovary, testis, subcutaneous tissue and kidney were found with a similar incidence in control and treated animals. Pulmonary adenocarcinomas were present in two male mice fed diets containing test chemical (one at 100 and one at 500 mg/Kg/day). Lymphosarcoma and reticulum-cell neoplasia had a similar incidence in mice fed test chemical and in the controls. Three squamous-cell carcinomas of the skin were seen in female mice (one control, one fed 10 mg/Kg/day and one fed 100 mg/Kg/day ). The other malignant tumours found were a fibrosarcoma in a control female, a rhabdomyosarcoma in a female given 10 mg/Kg/day test chemical and a mammary adenocarcinoma in a female given 100 mg/Kg/day of test chemical . Since no deviation was noted in the treated animals from the controls used, hence, the the no observed adverse effect level (NOAEL) for test chemical was considered to be 500 mg/kg bw/day.
Referenceopen allclose all
Table: Incidence of major tumors in rats fed test chemical
Tumor |
Dietary level |
||||
Controls |
250 |
500 |
1000 |
2500 |
|
Mammary adenocarcinoma |
2 |
2 |
2 |
- |
1 |
Mammary fibroadenoma 3 2 3 3 1 2 5 2 1 4 |
3 |
3 |
1 |
5 |
1 |
Pulmonary lymphosarcoma |
1 |
2 |
1 |
- |
1 |
Endometrial sarcoma |
- |
1 |
- |
- |
- |
Rats with tumors |
7 |
10 |
7 |
9 |
6 |
Table: Incidence of major tumors in rats fed test chemical
Tumor |
Dietary level |
||||
Controls |
250 |
500 |
1000 |
2500 |
|
Mammary adenocarcinoma |
2 |
2 |
2 |
- |
1 |
Mammary fibroadenoma 3 2 3 3 1 2 5 2 1 4 |
3 |
3 |
1 |
5 |
1 |
Pulmonary lymphosarcoma |
1 |
2 |
1 |
- |
1 |
Endometrial sarcoma |
- |
1 |
- |
- |
- |
Rats with tumors |
7 |
10 |
7 |
9 |
6 |
Table 1. Cumulative mortality of mice fedtest chemical at 0-3500 ppm of the diet for 80 wk
Week no |
Total no. of deaths |
|||||||
Males |
Females |
|
|
|
||||
0 |
70 |
700 |
3500 |
0 |
70 |
700 |
3500 |
|
10 |
0 |
0 |
0 |
0 |
0 |
1 |
0 |
0 |
20 |
0 |
3 |
0 |
0 |
0 |
1 |
1 |
1 |
30 |
0 |
3 |
0 |
0 |
0 |
1 |
1 |
1 |
40 |
1 |
3 |
0 |
0 |
0 |
1 |
3 |
1 |
50 |
1 |
4 |
1 |
0 |
0 |
3 |
4 |
4 |
60 |
3 |
6 |
2 |
1 |
4 |
7 |
7 |
5 |
70 |
9 |
10 |
4 |
4 |
9 |
11 |
10 |
9 |
80 |
12 |
12 |
10 |
9 |
17 |
18 |
16 |
16 |
Table 2. Mean body weights of mice fed Violet 6 B at 0-3500 ppm in the diet for 80 wk
Dose (ppm) |
Body weight (g) at week |
Weight gain (g) at wk 77 |
|||||
0 |
13 |
25 |
51 |
65 |
77 |
|
|
Males |
|||||||
0 |
28.1 |
34.0 |
34.3 |
47.6 |
42.6 |
43.0 |
14.9 |
70 |
27.8 |
33.7 |
35.2 |
45.2 |
42.1 |
42.0 |
14.2 |
700 |
26.3 |
31.8 |
32.0 |
43.3 |
41.2 |
40.6 |
14.3 |
3500 |
27.2 |
31.9 |
32.4 |
44.3 |
42.5 |
40.8 |
13.6 |
|
Females |
||||||
0 |
21.7 |
27.5 |
28.1 |
37.7 |
36.3 |
34.3 |
12.6 |
70 |
22.3 |
27.9 |
28.1 |
36.9 |
36.0 |
33.9 |
11.6 |
700 |
21.5 |
27.8 |
29.0 |
37.1 |
36.0 |
34.1 |
12.6 |
3500 |
21.7 |
28.4 |
28.4 |
37.0 |
36.0 |
35.9 |
14.2 |
Table 3. Absolute and relative organ weights of mice fed Violet 6B at 0-3500 ppm of the diet for 80 wk
Sex and dose (99m) |
No. of mice examined |
Organ weight |
Terminal body weight (g) |
||||||
Brain |
Heart |
Liver |
Spleen |
Kidney |
Stomach |
Small intestine |
|||
|
Absolute organ weight (g) |
||||||||
Males |
|
|
|
|
|
|
|
|
|
0 |
35 |
0.44 |
0.23 |
1.84 |
0.13 |
0.68 |
0.48 |
1.87 |
36 |
70 |
36 |
0.45 |
0.23 |
1.78 |
0.15 |
0.68 |
0.46 |
1.81 |
36 |
700 |
35 |
0.44 |
0.22 |
1.73 |
0.16 |
0.64 |
0.42 |
1.69* |
36 |
3500 |
38 |
0.44 |
0.22 |
1.73 |
0.14 |
0.65 |
0.43 |
1.65** |
36 |
Females |
|
|
|
|
|
|
|
|
|
0 |
30 |
0.45 |
0.18 |
1.66 |
0.17 |
0.49 |
0.52 |
1.80 |
30 |
70 |
29 |
0.44 |
0.17 |
1.72 |
0.17 |
0.50 |
0.49 |
1.68 |
30 |
700 |
27 |
0.48 |
0.18 |
1.68 |
0.21 |
0.51 |
0.47 |
1.78 |
31 |
3500 |
30 |
0.46 |
0.18 |
1.70 |
0.17 |
0.48 |
0.46 |
1.71 |
31 |
|
Relative organ weight (g/100 g body weight) |
||||||||
Males |
|
|
|
|
|
|
|
|
|
0 |
35 |
1.30 |
0.64 |
4.92 |
0.36 |
1.84 |
1.28 |
5.00 |
- |
70 |
36 |
1.22 |
0.61 |
4.79 |
0.44 |
1.83 |
1.19 |
4.74 |
- |
700 |
35 |
1.22 |
0.61 |
4.79 |
0.44 |
1.83 |
1.19 |
4.74 |
- |
3500 |
38 |
1.22 |
0.61 |
4.94 |
0.39 |
1.88 |
1.17 |
4.65 |
- |
Females |
|
|
|
|
|
|
|
|
|
0 |
30 |
1.50 |
0.60 |
5.51 |
0.57 |
1.63 |
1.66 |
6.02 |
- |
70 |
29 |
1.47 |
0.57 |
5.46 |
0.57 |
1.67 |
1.63 |
5.72 |
- |
700 |
27 |
1.55 |
0.58 |
5.47 |
0.68 |
1.68 |
1.52 |
5.77 |
- |
3500 |
30 |
1.48 |
0.58 |
5.56 |
0.55 |
1.57 |
1.49 |
5.58 |
- |
Table 4. Results of haematological studies in mice fed diets containing 0-3500 ppm Violet 6B for 80 wk
Sex and dose (ppm) |
No. of mice examined |
Hb (g/100 mL) |
PCV (%) |
RBC (103/mm3) |
Retics (% of RBC) |
Leucocytes |
||||
Total (103/mm3) |
Differential (%) |
|||||||||
N |
E |
L |
M |
|||||||
Males |
Week 52 |
|||||||||
0 |
10 |
14.0 |
44 |
7.65 |
1.9 |
8.94 |
16 |
0 |
83 |
1 |
700 |
10 |
13.9 |
47 |
7.97 |
- |
10.02 |
- |
- |
- |
- |
3500 |
10 |
14.9 |
47 |
8.41** |
1.6 |
11.55** |
19 |
0 |
81 |
0 |
Females |
|
|
|
|
|
|
|
|
|
|
0 |
10 |
14.6 |
46 |
7.41 |
1.8 |
7.44 |
23 |
0 |
73 |
1 |
700 |
10 |
14.3 |
49* |
6.84 |
- |
9.15 |
- |
- |
- |
- |
3500 |
10 |
14.3 |
47 |
7.31 |
2.1 |
7.72 |
18 |
0 |
81 |
1 |
Males |
Week 80 |
|||||||||
0 |
35 |
14.1 |
- |
- |
- |
- |
40 |
1 |
57 |
1 |
1 |
35 |
14.1 |
- |
- |
- |
- |
- |
- |
- |
- |
700 |
34 |
14.4 |
- |
- |
- |
- |
- |
- |
- |
- |
3500 |
37 |
14.2 |
- |
- |
- |
- |
37 |
1 |
61 |
1 |
Females |
|
|
|
|
|
|
|
|
|
|
0 |
30 |
15.4 |
- |
- |
- |
- |
50 |
1 |
48 |
1 |
70 |
28 |
14.9 |
- |
- |
- |
- |
- |
- |
- |
- |
700 |
29 |
14.7 |
- |
- |
- |
- |
- |
- |
- |
- |
3500 |
30 |
15.1 |
- |
- |
- |
- |
53 |
0 |
46 |
1 |
*P < 0.05; **P < 0.01.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 2 500 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
- Quality of whole database:
- Data is Klimicsh 2 and from authoritative database
Repeated dose toxicity: inhalation - systemic effects
Link to relevant study records
- Endpoint:
- repeated dose toxicity: inhalation, other
- Data waiving:
- other justification
- Justification for data waiving:
- other:
Reference
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: inhalation - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - systemic effects
Link to relevant study records
- Endpoint:
- repeated dose toxicity: dermal, other
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
Reference
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Repeated dose toxicity: Oral
Data available from the various test chemical was reviewed to determine the toxic nature of test chemical .The studies are as mentioned below:
Study 2.
Chronic repeated dose oral toxicity study was performed for test chemical to determine its toxic nature upon repeated exposure. The test chemical was mixed with feed at dose levels of0, 0.5, 1.0, 2.0 or 5.0 % (0, 250, 500, 1000 or 2500 mg/Kg bw/day) and fed for 2 years to 24 male and 24 female/dose Osborne Mendel weanling rats. During the study period, the animals were observed for mortality, any abnormality, body weight changes, hematology and were subjected to gross and microscopic pathology. No mortality and abnormality was noted during the study period. No significant growth inhibition occurred with the animals fed with test chemical . Also, no effects was observed on hemotological parameters and weights of heart, liver, kidney, spleen, and testis of the animals that received test chemical at dietary levels up to 5%(2500 mg/Kg bw/day). There was no treatment-related change in gross pathology. The usual gross pathologic conditions of old rats were seen in all groups, with no significant difference in distribution. The most common lesions were tumors, pneumonia (mostly chronic) of varying degrees? chronic nephritis of the usual type seen in our older rats, and testicular atrophy. There was no treatment-related change in microscopical examination. The most common tumors were mammary adenocarcinomas, fibroadenomas, and pulmonary lymphosarcomas. Based on the above considerations, the No observed Adverse Effect level (NOAEL) for test chemical was considered to be 2500 mg/kg bw/day in Osborne-Mendel male and female rats for 2 years feeding study.
Study 3.
Chronic repeated dose toxicity study was performed to determine the oral toxic nature of test chemical using ASH-CSI male and female mice. The test chemical was mixed with appropriate quantity of corn oil and mixed with feed at dose levels of 0, 10, 100 or 500 mg/kg/day. The animals were the test chemical in diet for 80 weeks and during the study the animals were observed for clinical signs, mortality, changes in body weight, hematology and the animals were subjected to gross and histopathology. The faeces of all treated mice were violet-coloured but the urine appeared normal, suggesting that the dose was largely unabsorbed from the gastro-intestinal tract. The feeding of Violet 6B did not affect the mortality, the rate of body-weight gain or the haematological picture at 12, 28, 52 or 80 wk. The only statistically significant changes of organ weight were in the small intestine of the male mice. Those given diets containing 100 or 500 mg/Kg/day were lower, by 10 and 12% respectively, than the controls. This effect was not statistically significant when the weights were expressed relative to body weight. Most of the lesions were those normally occurring in ageing mice and were found with a similar incidence in treated and control mice and appeared to be not related to the treatment with test chemical . The changes found in the skin consisted of hyperkeratosis, also present in control animals, and a single dermoid cyst in a female given 700 ppm test chemical Dilated gastric glands and hyperplasia of the gastric mucosa were encountered, but again similar lesions were found in control mice. Pulmonary adenomas were seen in 36% of all male mice and in 20% of all female mice, the incidence being similar in all groups. Two thyroid adenomas were found in male mice fed test chemical (one at 10 and the other at 500 mg/Kg/day). Benign tumours of the ovary, testis, subcutaneous tissue and kidney were found with a similar incidence in control and treated animals. Pulmonary adenocarcinomas were present in two male mice fed diets containing test chemical (one at 100 and one at 500 mg/Kg/day). Lymphosarcoma and reticulum-cell neoplasia had a similar incidence in mice fed test chemical and in the controls. Three squamous-cell carcinomas of the skin were seen in female mice (one control, one fed 10 mg/Kg/day and one fed 100 mg/Kg/day ). The other malignant tumours found were a fibrosarcoma in a control female, a rhabdomyosarcoma in a female given 10 mg/Kg/day test chemical and a mammary adenocarcinoma in a female given 100 mg/Kg/day of test chemical . Since no deviation was noted in the treated animals from the controls used, hence, the the no observed adverse effect level (NOAEL) for test chemical was considered to be 500 mg/Kg/day.
Based on the data available, the No Observed Adverse Effect Level (NOAEL) for the test chemical was considered to be in range of 500 -2500 mg/Kg bw using male and female rats or mice . Hence the test chemical is not likely to be toxic upon repeated exposure by oral route as per the criteria mentioned in CLP regulation.
Repeated dose toxicity: inhalation
The melting point of test chemical is >300 °C.This suggests that test chemical decomposes above300 °C without melting and also it does not exhibit very high vapour pressure. Inhalation is therefore not the likely route of exposure and hence this end point is considered for waiver.
Repeated dose toxicity: dermal
Repeated dermal toxicity is unlikely to occur since dermal absorption of test chemical is limited. Also considering the use of the chemical as a colouring agent in food and cosmetics and considering the volatility absorption by the dermal route is not considered to be significant. Thus, given the above considerations, it is assumed that test chemical shall not exhibit acute dose toxicity by the repeated dermal route.
Justification for classification or non-classification
Based on the data available and applying the weight of evidence approach, the test chemical does not exhibit toxic nature upon repeated exposure by oral route of exposure. Hence the test chemical is likely to be not classified for toxicity upon repeated dose exposure by oral, dermal and inhalation route of exposure.
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