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Reaction mass of pentasodium 2-{[4-chloro-6-(ethyl{3-[(2-sulfonatoethyl)sulfonyl]phenyl}amino)-1,3,5-triazin-2-yl]amino}-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate and tetrasodium 2-[(4-chloro-6-{ethyl[3-(vinylsulfonyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-5-hydroxy-6-({2-sulfonato-4-[(4-sulfonatophenyl)diazenyl]phenyl}diazenyl)naphthalene-1,7-disulfonate
EC number: 459-580-2 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 27 January 2006 to 19 June 2006
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 006
- Report date:
- 2006
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: Directive 96 / 54 / EC, B. 7. "Repeated dose (28 days) toxicity (oral)", September 30, 1996.
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: Japanese Guidelines for Screening, Toxicity Testing of Chemicals: Testing Methods for new Substances, enacted July 13, 1974, amended December 5, 1986.
- Deviations:
- not specified
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Test material form:
- solid: particulate/powder
- Details on test material:
- -Batch No.: ROE 805 BOP 04/05
-Colour: Dark red
-Solubility in water: >100g/L at 20 °C
-Solubility in vehicle: Miscible
-Stability in solvent: 7 days in Water, Saline, Polyethylene Glycol, Carboxymethylcellulose, and 1 day in Vaseline and FCA at room temperature.
-Storage: At room temperature, in the desicator
-Expiration Date: October 01, 2010
-Purity: Approx. 82 % organic part (Na-salt),
all coloured components = 80.3 %;
Main component 1: 36.2 %,
Main component 2: 27.5 %, Oligomers: 10 %
Constituent 1
- Specific details on test material used for the study:
- Identity: FAT 40824/A
Batch: Red ROE 805 BOP 04/05
Appearance: dark red powder
Purity: Organic part (Na-salt): approx. 82 %; Main component 1 : approx. 36.2 %; Main component 2: approx. 27.5 %; Oligomers: 10%
Expiration date: 01 October 2010
Storage: At room temperature at about 20°C in a desiccator (hygroscopic)
Test animals
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
Test system: Rat, HanRcc:WIST (SPF)
Source: RCC Ltd Laboratory Animal Services CH-4414 Füllinsdorf / Switzerland
Total number of animals per group: Groups 1 and 4: each 10 males and 10 females, Groups 2 and 3: each 5 males and 5 females
Total number of animals used: 30 males and 30 females
Age at delivery: 6 weeks
Body weight range (acclimatization): Males: 136 - 159 grams (mean 146 grams), Females: 116 - 133 grams (mean 123 grams)
Acclimatization: Under test conditions after health examination. Only animals without any visible signs of illness were used for the study.
Water: Community tap-water from Itingen was available ad libitum in water bottles.
Diet: Pelleted standard Provimi Kliba 3433 (batch no. 76/05) rat maintenance diet (Provimi Kliba AG, CH- 4303 Kaiseraugst/ Switzerland) was available ad libitum.
ENVIRONMENTAL CONDITIONS
Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environmental conditions (temperature range: 22 ±3 °C; relative humidity range: 30- 70 %). There was 12-hour fluorescent light/12-hour dark cycle with music during the light period.
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks:
- Bidistilled water
- Details on oral exposure:
- Rationale for dose level selection: Based upon the results of a non-GLP 5-day doserange-finding study (RCC Study Number A45325) in which FAT 40824/A was administered by gavage to 2 rats per group and sex.
Dose volume: 10 ml/kg body weight
Duration of acclimatization period: 7 days
Duration of recovery: 14 days - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Concentration, homogeneity and stability (after 2 hours and 7 days) of the dose formulations were determined in samples taken after experimental start. Concentration and homogeneity of the dose formulations were determined in samples taken during week 3 of the treatment. The analyses were performed by RCC Ltd.
- Duration of treatment / exposure:
- 28 days
- Frequency of treatment:
- Daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- Group 1 (Control)
- Dose / conc.:
- 50 mg/kg bw/day (actual dose received)
- Remarks:
- Group 2 (Low dose)
- Dose / conc.:
- 200 mg/kg bw/day (actual dose received)
- Remarks:
- Group 3 (Middle dose)
- Dose / conc.:
- 1 000 mg/kg bw/day (actual dose received)
- Remarks:
- Group 4 (High dose)
- No. of animals per sex per dose:
- Groups 1 and 4: each 10 males and 10 females,
Groups 2 and 3: each 5 males and 5 females - Control animals:
- yes, concurrent vehicle
- Details on study design:
- Dose Formulation: The dose formulations were prepared weekly. FAT 40824/A was weighed into a tared glass beaker on a tared Mettler balance and the vehicle added (weight:volume). The mixtures were prepared using a magnetic stirrer and stored at room temperature (15-25 °C). Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
- Positive control:
- no data
Examinations
- Observations and examinations performed and frequency:
- Mortality / Viability: Observations for mortality/viability were recorded twice daily.
General Cageside Observations (Daily): The animals were observed for clinical signs once before commencement of administration; twice daily on days 1-3; as well as once daily on days 4-28, and once daily during days 29-42 (recovery).
Detailed Clinical Observations (Weekly): The animals were observed in their home cages, outside their home cages in a standard arena and in the hand. These observations were performed in random sequence once before commencement of administration and once weekly (weeks 1-3) thereafter.
Food Consumption: The food consumption was recorded once during the acclimatization and weekly thereafter, using an on-line electronic recording system consisting of a Mettler balance connected to the RCC computer.
Body Weights: Body weights were recorded weekly during acclimatization, treatment and recovery periods and before necropsy, using an on-line electronic recording system consisting of a Mettler balance connected to the RCC computer.
Functional Observational Battery: During treatment week 4, relevant parameters from a modified Irwin screen test were evaluated in all animals.
Clinical Laboratory Investigations: Blood samples were drawn from the retro-orbital plexus from all animals under light
isoflurane anesthesia. The animals were fasted in metabolism cages for approximately 18 hours before blood sampling but allowed access to water ad libitum. The samples were collected early in the working day to reduce biological variation caused by circadian rhythms. Urine was collected during the 18 hours fasting period into a specimen vial, using a metabolism cage. - Sacrifice and pathology:
- Sacrifice: after 4 weeks 03 March 2006 (Allocation A), after 6 weeks 17 March 2006 (Allocation B)
All animals were weighed and necropsied. Descriptions of all macroscopic abnormalities were recorded. All animals were anesthetized by intraperitoneal injection of sodium pentobarbitone and killed by exsanguination. Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered 4 % formaldehyde solution. - Other examinations:
- The following organ weights were recorded on the scheduled dates of necropsy: Brain, Thymus, Spleen, Ovaries, Heart, Kidneys, Testes, Liver, Adrenals, Epididymides.
The organ to terminal body weight ratios as well as organ to brain weight ratios were determined. The determination of the terminal body weight was performed immediately prior to necropsy.
Histotechnique: All organ and tissue samples, as defined under Histopathology (following), were processed, embedded and cut at an approximate thickness of 2 to 4 micrometers, and stained with hematoxylin and eosin.
Histopathology: Slides of all organs and tissues listed in boldface type (see Necropsy, above) that were collected at scheduled sacrifice from the animals of control and high-dose groups were examined by a pathologist. - Statistics:
- The following statistical methods were used to analyze the grip strength, locomotor activity, body weight, body weight gain, organ weights and ratios, clinical laboratory investigations and macroscopic findings as well as:
• The Dunnett-test (many to one t-test) based on a pooled variance estimate were applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test was applied to the macroscopic findings.
• Student’s T-test was applied to locomotor activity and grip strength.
Results and discussion
Results of examinations
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- Dark red feces were observed in rats treated with 1000 mg/kg body weight/day and 200 mg/kg body weight/day from treatment day 3 onwards. Dark red feces were present in rats treated with 1000 mg/kg body weight/day during recovery period until day 5. This finding is considered to be a passive finding related to the oral administration of large doses of a dyestuff and without toxicological relevance.
At 200 mg/kg body weight/day, crusts on the left shoulder were observed each in one male during treatment weeks 2 and 3 and a kinked tail apex was noted during treatment week 3. These changes are considered to be test item unrelated. - Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Description (incidence and severity):
- No test item-related changes were noted at any dose level in the mean body weights.
- Food consumption and compound intake (if feeding study):
- no effects observed
- 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):
- However, incidental changes without toxicological relevance were noted in the hematology parameters after 4 weeks’ treatment in the rats as follows:
Platelet counts were significantly decreased in males treated with 50 mg/kg body weight/day (p<0.01) or 200 mg/kg body weight/day (p<0.05). In females the mean platelet count was reduced (not significant) at 200 mg/kg body weight/day and increased at 1000 mg/kg body weight/day (p<0.05). In the absence of a clear doseresponse relationship, these changes are considered to be incidental.
At 50 mg/kg body weight/day in males only, decreased relative monocytes (p<0.05) and leukocyte counts (not significant) were recorded.
At 200 mg/kg body weight/day in males only, significantly increased relative reticulocyte counts (p<0.05) were noted when compared with their respective controls.
At 1000 mg/kg body weight/day only in females, significantly decreased hemoglobin (p<0.05) and mean corpuscular hemoglobin (p<0.01) concentrations but elevated prothrombin time (p<0.05) were noted.
All of the statistically significant differences remained within the ranges of the historical control data and were considered to be incidental.
A few test item related late effects were noted after 2 weeks’ recovery.
The following hematology parameters were significantly increased in males:
erythrocytes count (p<0.05), hemoglobin concentration (p<0.05), hematocrit (p<0.05), relative (p<0.05) and absolute (p<0.01) basophils and monocytes (p<0.05). In males, red cell volume distribution width was significantly decreased (p<0.05).
In females, only the leukocyte count was significantly increased (p<0.05).
All observed changes in the hematology parameters after treatment and recovery periods remained within the range of the historical control data and are considered to be incidental. - Clinical biochemistry findings:
- no effects observed
- Description (incidence and severity):
- However, at 1000 mg/kg body weight/day after treatment and recovery periods, blood plasma was reddish discolored. This finding is considered to be a passive finding related to the oral administration of large doses of a dyestuff and without toxicological relevance.
The following changes in the clinical biochemistry parameters were noted after 4 weeks of treatment:
At 1000 mg/kg body weight/day, total bilirubin concentration was significantly elevated (p<0.05 in both genders) out of the 95% tolerance limits. It is considered to be test item (dyestuff) related passive effect.
Concentrations of cholesterol and phospholipids were elevated (p<0.05 both) in males treated with 50 mg/kg body weight/day. At 1000 mg/kg body weight/day in both genders, triglycerides (not significant) were increased. At 1000 mg/kg body weight/day in females only, cholesterol was increased (p<0.05). These differences remained within the ranges of the 95% tolerance limits and are considered to be incidental.
Creatine kinase activity was changed in test item treated rats (50 / 200 / 1000 mg/kg body weight/day) in males (+89 / +60 / +35 %) and in females (+78 / -53 / -45 %) without statistically significance, which exceeded the higher range of the 95% tolerance limits. This finding is considered to be test item unrelated, as no dose response relationship could be established.
A number of test item unrelated changes were noted after 4 weeks’ treatment in the clinical biochemistry parameters of rats treated with 1000 mg/kg/day: reduced activities of alanine aminotransferase (p<0.05 in females only) and alkaline phosphatase (not significant, in females only), but increased activity of lactate dehydrogenase (p<0.05 in males only). The latter finding is considered to be the result of plasma discoloration (i.e.
an artifact).
Potassium levels were increased in males at all dose levels (not significant) and in females at 50 mg/kg body weight/day (p<0.05). Phosphorus levels were increased in males (p<0.05) at 200 mg/kg body weight/ day and in females (p<0.05) at 50 mg/kg body weight/day.
These changes were not dose related and are considered to be incidental.
After 2 weeks’ recovery, increased phospholipids (p<0.05), increased creatine kinase activity (p<0.05) and reduced chloride level (p<0.05) were noted in females when compared with the controls. Not significant changes were noted in increased cholesterol and triglycerides, reduced activities of alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase and glutamate dehydrogenase. All differences remained within
the 95% tolerance limits of the historical control data and are considered to be incidental. - Urinalysis findings:
- no effects observed
- Description (incidence and severity):
- No changes of toxicological relevance were noted in the urinary parameters in rats of both sexes at any dose level.
At 1000 mg/kg body weight/day, urine pH-values were elevated in males (not significant) and females (p<0.05) and nitrite was increased (p<0.05) in females only.
Erythrocytes were elevated in males treated with 1000 mg/kg body weight/day (p<0.05), but remained within the ranges of the historical control data and is considered to be incidental.
After 2 weeks’ recovery, increased concentrations of protein, erythrocytes and leukocytes were noted in males and decreased protein concentration in females. All changes were not significant and remained within the historical control data excepting erythrocytes in males. - Behaviour (functional findings):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- No differences of toxicologically relevance in mean absolute and relative organ weights were noted after treatment and treatment-free recovery period in male and female rats at any dose level.
However few incidental changes observed as follows
After 4 Weeks
No toxicologically relevant differences in mean absolute and relative organ weights were noted after a 4-week treatment period in male and female rats at any dose level. In males treated with 50 and 200 mg/kg body weight/day, slightly elevated (+6 % and +8 %) testes weights were noted when compared with control. In females treated with 200 mg/kg body weight/day, decreased (-14 %) ovary weights were noted. In the absence of a dose response relationship, these findings are considered to be incidental.
After 6 Weeks
No toxicologically relevant differences in mean absolute and relative organ weights were noted after a two-week treatment-free recovery period in male rats.
Following differences were noted in females when compared with their respective controls: increased absolute kidneys (p<0.05) weights, increased liver- (p<0.01) and kidneys-to-body weight (p<0.01) ratios, increased liver- (p<0.05) and kidneys-to-brain weight ratios (p<0.05). No toxicologically relevant differences were noted after the 4- weeks of treatment and therefore these changes are considered to be incidental. - Gross pathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- After 4 Weeks
In all rats treated with 1000 mg/kg body weight/day, reddish discoloration of the mucosa of the rectum was noted at necropsy. Reddish discoloration of the kidneys was observed in females treated with 1000 mg/kg body weight/day. These observations are considered to be test item (dyestuff-) related.
Further macroscopic findings recorded (renal pelvis dilation or isolated reddish foci on stomach fundus) are considered to be within the range of the normal background and biological variance.
After 6 Weeks
No macroscopic findings of toxicological relevance were noted in any rats after a two week treatment-free recovery period.
Following macroscopic findings were recorded in males: dark red foci on seminal vesicles and stomach fundus and dark red discolored renal lymph nodes. Findings recorded are considered to be test item-related but characteristic effects caused by administration of a dyestuff. - Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- At the end of the treatment period, reddish discoloration of the rectum (males and females) and kidneys (females only) was recorded in animals treated with 1000 mg/ kg body weight/day). The microscopic examination of these organs did not reveal any changes associated to this finding and, therefore, no toxicological relevance was attributed to them.
At microscopic examination performed at the end of the treatment and following recovery period no test item-related microscopic lesions were observed.
Under the conditions of this experiment, the test item FAT 40824/A produced no morphological evidence of toxicological properties in the organs and tissues examined. - Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
- Details on results:
- FUNCTIONAL OBSERVATIONAL BATTERY
No test item-related findings were noted during the functional observational battery (treatment week 4).
GRIP STRENGTH
No test item-related alterations in the mean fore- or hindlimb grip strength were noted at any dose level
LOCOMOTOR ACTIVITY
No test item-related differences in mean locomotor activity were noted at any dose level.
Effect levels
open allclose all
- Dose descriptor:
- NOEL
- Effect level:
- 200 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: reddish discoloration of kidneys (females only) and rectum (both genders) at high dose group (1000 mg/kg bw/d), which is considered treatment related but of no toxicological relevance as the substance is a dyestuff (colourant).
- Dose descriptor:
- NOAEL
- Effect level:
- 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: Overall study results
Target system / organ toxicity
- Critical effects observed:
- not specified
Applicant's summary and conclusion
- Conclusions:
- The no-observed-adverse-effect-level (NOAEL) and the no-observed-effect-level (NOEL) of FAT 40824/A in 28 day repeated dose toxicity study was 1000 and 200 mg/kg bw/day respectively.
- Executive summary:
A GLP compliant 28 day repeated dose toxicity by oral administration of FAT 40824/A to Wistar rats at doses of 50, 200 and 1000 mg/ kg bw/day was carried out according to OECD guideline 407. A control group was treated similarly with the vehicle, bidistilled water, only. The groups comprised 5 animals per sex, which were sacrificed after 28 days of treatment. Additional 5 rats per sex and group were used at 0 and 1000 mg/kg body weight/day. These animals were treated for 28 days and then allowed a 14-day treatment-free recovery period after which they were sacrificed. Clinical signs, outside cage observation, food consumption and body weights were recorded periodically during acclimatization, and during the treatment and recovery periods. Functional observational battery, locomotor activity and grip strength were performed during week 4. At the end of the dosing and the treatment-free recovery period, blood samples were withdrawn for hematology and plasma chemistry analyses. Urine samples were collected for urinalyses. All animals were killed, necropsied and examined post mortem. Histological examinations were performed on organs and tissues from all control and high dose animals, and all gross lesions from all animals.
There were no mortality, no clinical signs of toxicological relevance during the weekly observations (treatment weeks 1-3) or during functional observational battery (treatment week 4), no effects on grip strength or locomotor activity, no effects on mean daily- or relative food consumption, no effects on body weight and body weight gain, no effects on hematology, clinical biochemistry or urinalysis, no effects on mean absolute or relative organ weights and no macroscopic findings of toxicological relevance. The presence of dark feces during the treatment period and during the first days of the recovery period noted in rats treated with 1000 mg/kg body weight/day was considered to be a passive finding without toxicological relevance.
Based on the results of this study, the no-observed-adverse-effect-level (NOAEL) and the no-observed-effect-level (NOEL) of FAT 40824/A in 28 day repeated dose toxicity study was 1000 and 200 mg/kg bw/day respectively.
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.
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