Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
Reaction mass of 5,5'-{(phenylmethanediyl)bis[benzene-4,1-diyl-diazene-2,1-diyl]}bis{1-[3-(dimethylamino)propyl]-4-methyl-6-oxo-3-(pyridinium-1-yl)-1,6-dihydropyridin-2-olate} hydrochloride and 5,5’-[3,4’-(phenylmethanediyl)diphenylene]bis(diazene-2,1-diyl)bis{1-[3-(dimethylamino)propyl]-4-methyl-6-oxo-3-(pyridinium-1-yl)-1,6-dihydropyridin-2-olate} hydrochloride
EC number: 700-312-3 | 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
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 2007-12-13 to 2008-04-25
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline-conform study under GLP without deviations, conducted with the analogue substance.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 008
- Report date:
- 2008
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:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- analogue substance (refer to IUCLID chapter 13)
- IUPAC Name:
- analogue substance (refer to IUCLID chapter 13)
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: RCC Ltd Laboratory Animal Services, Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
- Age at study initiation: Approx. 6 weeks
- Weight at study initiation: Males: 124.1 to 158.3 g (141.3 ± 8.8 g); Females: 111.3 to 132.3 g (122.9 ± 5.1 g)
- Fasting period before study: not reported
- Housing: In groups of five in Makrolon type-4 cages with wire mesh tops and sterilized standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard rat maintenance diet was available ad libitum.
- Water (e.g. ad libitum): Community tap-water from Itingen was available ad libitum in water bottles.
- Acclimation period: 7 days, under test conditions
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12-hour fluorescent light/12-hour dark cycle
IN-LIFE DATES: From: 2007-12-14 To: 2008-01-18 (main group), 2008-02-01 (recovery group)
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- water
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
The dose formulations were prepared weekly. Test item was weighed into a glass beaker on a tared Mettler balance and the vehicle, purified water, was added to give the appropriate final concentration of test item in the suspension. The mixtures were prepared using a magnetic stirrer Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
DIET PREPARATION
- not applicable
VEHICLE
- Justification for use and choice of vehicle (if other than water): water
- Concentration in vehicle: 5, 15, and 45 mg/l
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): not applicable
- Purity: purified water was used - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- General procedures:
On the first treatment day (21-Dec-2007) and once during week 3 (04-Jan-2008), one sample of the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of homogeneity and concentration. One sample of about 2 g of each concentration was taken 2 hours and 7 days after commencement of dosing (dose formulation of the first treatment day) to confirm 2 hrs and 7 d-stability at room temperature (20 ± 5 °C). The aliquots for analysis of dose formulations were delivered to Dr. D. Flade (RCC Ltd, Itingen / Switzerland) and stored at -20 ± 5 °C until analysis. The dose formulations were delivered under ambient conditions. Analyses were be performed by the Study Scientist, Dr. D. Flade, according to an HPLC analytical method supplied by the Sponsor and previously adapted at RCC Ltd (RCC Study No.
B62032). The test item served as analytical standard. Details of the analytical method were documented in the raw data and in the part report generated by the Study Scientist.
Analytical Procedure
Preparation of Standard Solutions:
Stock solutions of test item in acetonitrile/purified water (1+1 v/v) were prepared for external standard calibration. For example, 25.29 mg of the test item was exactly weighed into a 50 mL volumetric flask and approximately 40 mL of acetonitrile/purified water (1+1 v/v) was added. Then, the mixture was sonicated for 5 minutes and the flask was brought to volume with acetonitrile/purified water (1+1 v/v) to yield a solution with a concentration of 505.8 μg/mL. Aliquots of this stock standard solution were used to prepare five working standard solutions in acetonitrile/purified water (1+1 v/v) with a concentration range of 10.12 to 101.2 μg/mL. Thirteen standard solutions derived from two stock standard solutions were used for calibration.
Analysis of Samples:
The samples received were dissolved in acetonitrile/purified water (1+1 v/v) by sonication for 5 minutes and then diluted to volume with acetonitrile/purified water (1+1 v/v). The sample solutions were further diluted with acetonitrile/purified water (1+1 v/v) into the calibration range.
High Performance Liquid Chromatographic Determination:
Apparatus: Pump: Merck-Hitachi L -7100
Sampling Unit: Merck-Hitachi L-7200
Column Oven: Merck-Hitachi L-7300
UV/VIS Detector: Merck-Hitachi L-7400
Interface: Merck-Hitachi D-7000
Column: Symmetry C18; 150 x 3.9 mm, 5 μm
Pre-Column: Phenomenex C18; 4 x 3 mm, 5 μm
Column Temperature: 40 °C
Eluent A: 0.1% Trifluoro acetic acid in purified water
Eluent B: Acetonitrile
Gradient: Eluent A 65%, Eluent B 35 % (at 0 and 6 min, resp.)
Flow: 1.0 mL/min
Wave Length: 254 nm
Injection Volume: 10 μL
Evaluation of Results
Injected samples were quantified by comparing peak areas of the test item with reference to the calibration curve. The latter was obtained by correlation of the peak areas of the working standards with their corresponding concentrations (μg/mL), using linear regression. - Duration of treatment / exposure:
- 28 days,
plus 14 days untreated recovery period of control and high dose animals - Frequency of treatment:
- 7 days/week
Doses / concentrationsopen allclose all
- Remarks:
- Doses / Concentrations:
0 mg/kg bw/d
Basis:
actual ingested
- Remarks:
- Doses / Concentrations:
50 mg/kg bw/d
Basis:
actual ingested
- Remarks:
- Doses / Concentrations:
150 mg/kg bw/d
Basis:
actual ingested
- Remarks:
- Doses / Concentrations:
450 mg/kg bw/d
Basis:
actual ingested
- No. of animals per sex per dose:
- Males: 10 animals at 0 mg/kg bw/day; 5 animals at 50 mg/kg bw/day; 5 animals at 150 mg/kg bw/day; 10 animals at 450 mg/kg bw/day.
Females: 10 animals at 0 mg/kg bw/day; 5 animals at 50 mg/kg bw/day; 5 animals at 150 mg/kg bw/day; 10 animals at 450 mg/kg bw/day. - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The dose levels were selected based on a previous dose range finding toxicity study in Wistar rats, RCC Study Number B62032, using dose levels of 100, 250, and 500 mg/kg/day.
- Rationale for animal assignment (if not random): Computer-generated random algorithm.
- Rationale for selecting satellite groups: no data
- Post-exposure recovery period in satellite groups: 14 days
- Section schedule rationale (if not random): random - Positive control:
- no
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Observations for viability / mortality were recorded twice daily. The animals were observed for clinical signs once daily during acclimatization, twice daily on treatment days 1-3 and once daily thereafter including the recovery period.
- Cage side observations checked in table 1, attached below, were included.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly
BODY WEIGHT: Yes
- Time schedule for examinations: Body Weights were recorded weekly during acclimatization, treatment and recovery periods and before necropsy.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- The food consumption was recorded once during the acclimatization period and weekly thereafter (however no feeding study)
FOOD EFFICIENCY:
- No
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 4 Weeks: main group and recovery group. After 6 Weeks: recovery group
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all involved in the study
- Parameters checked in table 2, below, were examined.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 4 Weeks: main group and recovery group. After 6 Weeks: recovery group
- Animals fasted: Yes
- How many animals: all involved in the study
- Parameters checked in table 2, below, were examined.
URINALYSIS: Yes
- Time schedule for collection of urine: After 4 Weeks: all animals. After 6 Weeks: recovery group
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table 2, below, were examined.
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: During week 4
- Dose groups that were examined: all animals examined
- Battery of functions tested: sensory activity / grip strength / motor activity / - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
Adrenal glands
Aorta
Bone (sternum, femur including joint)
Bone marrow (femur)
Brain – including section of medulla/pons,
cerebral and cerebellar cortex
Cecum
Colon
Duodenum
Epididymides (fixed in Bouin's solution)
Esophagus
Eyes w/optic nerve (fixed in Davidson's
solution)
Harderian gland (fixed in Davidson's solution)
Heart including auricles
Ileum, with Peyer's patches
Jejunum with Peyer's patches
Kidneys
Larynx
Lacrimal gland, exorbital
Nasal cavity
Ovaries
Pancreas
Pharynx
Pituitary gland
Prostate gland incl. coagulating glands
Rectum
Salivary glands - mandibular, sublingual
Sciatic nerve
Seminal vesicles
Skeletal muscle
Skin
Spinal cord - cervical, midthoracic, lumbar
Spleen
Stomach
Testes (fixed in Bouin's solution)
Thymus
Thyroid (incl. parathyroid gland, if possible)
Tongue
Trachea
Liver
Lungs, filled w/formalin at necropsy
Lymph nodes - mesenteric, mandibular and
popliteal
Mammary gland area
Urinary bladder, filled w/formalin at
necropsy
Uterus
Vagina
Gross lesions
HISTOPATHOLOGY: Yes (all organs as stated above) - Statistics:
- The following statistical methods were used to analyze the grip strength, locomotor activity, body weight, macroscopical findings, organ weights and ratios, as well as clinical laboratory data:
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was 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
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Mortality occurred in one male and one female of the 150 mg/kg/d group, and in three males and two females of the 450 mg/kg/d group due to incidental aspiration of test material caused by the oral gavage application. Mortalities are considered not related to systemic toxicity of the test substance.
- Mortality:
- mortality observed, treatment-related
- Description (incidence):
- Mortality occurred in one male and one female of the 150 mg/kg/d group, and in three males and two females of the 450 mg/kg/d group due to incidental aspiration of test material caused by the oral gavage application. Mortalities are considered not related to systemic toxicity of the test substance.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- 450 mg/kg/day (males)
- Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- 450 mg/kg/day (males)
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- 150 mg/kg/day: Leukocytosis was recorded in females; in both sexes at 450 mg/kg/day (fully reversible at 150 mg/kg bw/d, partly reversible at 450 mg/kg/day).
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- 150 mg/kg/day: females cholesterol levels were slightly to moderately increased (partly reversible).
- Urinalysis findings:
- effects observed, treatment-related
- Description (incidence and severity):
- 150 mg/kg/day: marked increases in protein concentration and leukocyte count were noted in both sexes; erythrocyte counts were increased in females (all effects reversible).
- Behaviour (functional findings):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- 450 mg/kg/day: Increased liver, kidney and spleen weights recorded in females (reversible).
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- discoloration effects all reversible
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- 150 and 450 mg/kg/day: increase in incidence and mean severity grade (minimal to moderate) of hyaline droplets in the kidneys of males (not classification relevant, as effect to be considered as not relevant for humans).
- Histopathological findings: neoplastic:
- no effects observed
Effect levels
- Dose descriptor:
- NOAEL
- Effect level:
- 450 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: Based on an oral (28-d) subacute study with the structural closely related source molecule a NOAEL of 450 mg/kg bw/d is delineated for the registration substance as target molecule using read-across.
Target system / organ toxicity
- Critical effects observed:
- not specified
Applicant's summary and conclusion
- Conclusions:
- Based on the oral (28-d) subacute study with the structural closely related source molecule a NOAEL of 450 mg/kg bw/d is delineated for the registration substance as target molecule using read-across.
- Executive summary:
A valid oral (28-day) subacute repeated dose toxicity study was conducted with the analogue substance. The applicability of the read across approach was confirmed by the registrant via an expert statement (refer to IUCLID chapter 13), and is based on very close structural comparability as well as the very close physico-chemical characteristics of target and source molecule.
Findings related to the treatment with the analogue substance consisted of unspecific clinical symptoms like breathing noises and ruffled fur noted in high dose animals, and a slight but significant decrease in body weights of male animals of the high dose group which, however, reversed during the recovery period. Haematology revealed a minor degree of microcytar and hyperchromatic anemia which, however, were not accompanied by histologic lesions in the bone marrow. According to the pathologist, the observed increase in reticulocytes indicated a compensatory effect to changes in the hemoglobin synthesis that was fully reversible during the recovery period. Clinical chemistry revealed slightly to moderately increased cholesterol levels in male and female animals. Additionally, slight changes in urine parameters were also reversible after the recovery period.
With regard to organ weights, slight but significant increased absolute and relative liver, kidney and spleen weights in females of the high dose group were recorded, which were all reversible during recovery. A dose-response relationship was not seen and no histopathological correlate indicative of structural and/or functional changes was identified in any of these organs. No significant organ weight changes were observed in females of the low and mid dose group and in males of any dose group.
Pigment related yellowish-brown discoloration was noted in some organs like trachea, tongue, esophagus and gastrointestinal tract. However, although pigment was observed in tissues like trachea or intestinal mucosa, no degenerative lesions were observed by histopathology in these or any other organ investigated.
Microscopically an increase in incidence and mean severity grade from minimal to moderate of hyaline droplets in the kidneys of mid and high dose male animals was observed. However, no histopathological correlate in the kidneys in the affected animals was found. Moreover, these findings were reversible during recovery and no difference was seen to the animals from the control group. It is scientifically well established, that the male rat is prone to hyaline droplet formation in the kidneys and that this unique male rat-specific kidney effect has no relevance for human health (Flamm and Lehman-McKeeman 1991; Lehmann-McKeeman and Caudill 1992; Swenberg 1993; Hard eta al. 1993).
Based on the study results and in depth pathological evaluations, it was concluded by the study director that all findings are not considered adverse in nature but mainly of functional adaptive character. These effects were all reversible in a subsequent recovery period. The observed mortalities are pathologically primarily related to incidental aspiration of test material and not considered to represent systemic toxicity. Based hereupon it was concluded that the NOAEL in this study can be determined to be 450 mg/kg body weight per day.
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.