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EC number: 423-300-7 | CAS number: 128554-52-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- Experimental starting date: 17 April 2007; Experimental completion date: 20 July 2007
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Study data over 12 years old provided by ECHA, on a previously notified substance considered comparable and suitable for read-across use for the substance being registered (see attachments for justification of read-across). Study was performed according to OECD and EU guidelines and according to GLP principles.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 007
- Report date:
- 2007
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: EC Directive 67/548/EEC, B Repeated Dose (90 days) Toxicity (oral), 2001
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Japanese Chemical Substances Control Law 1987, Notification of Nov.21 2003 by MHLW (No. 1121002), METI (No.2) and ME (No. 031121002)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- -
- EC Number:
- 423-300-7
- EC Name:
- -
- Cas Number:
- 128554-52-9
- Molecular formula:
- Molecular formula varies for each reaction product. See section 1.2 for the molecular formula for each identified reaction product.
- IUPAC Name:
- 12-hydroxy-N-({3-[(12-hydroxyoctadecanamido)methyl]phenyl}methyl)octadecanamide
- Reference substance name:
- AMIDE#71
- IUPAC Name:
- AMIDE#71
- Details on test material:
- - Name of test material (as cited in study report): AMIDE #71
- Substance type: White powder
- Physical state: solid
- Stability under test conditions: Stable
- Storage condition of test material: At room temperature in the dark
Constituent 1
Constituent 2
Test animals
- Species:
- rat
- Strain:
- Wistar
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 6 weeks
- Weight at study initiation: At Day 1 the group mean weight for males was 216 - 220 g and for females 170 - 171 g. All animals within ± 20% of the sex mean.
- Fasting period before study: None.
- Housing: Group housing of 5 animals per sex in Macrolon cages with sterilised sawdust as bedding material and paper as cage-enrichment. No cage-enrichment was provided during overnight activity monitoring.
- Diet (e.g. ad libitum): Free access to pelleted diet.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: At least five days before the start of treatment under laboratory conditions.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): Optimal conditions were considered to be approximately 21 ± 3.0°C (actual range: 20.3 - 24.1°C)
- Humidity (%): Optimal conditions were considered to be approximately 30-70% (actual range: 33-94%)
- Air changes (per hr): Approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- propylene glycol
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
Vehicle: Propylene glycol (specific gravity 1.036)
Rationale for vehicle: Based on trial formulations prepared and on information from the sponsor.
Method of formulation: Formulations (w/w) were prepared daily within 5 hours prior to dosing and were homogenised to visually acceptable levels. Adjustment was made for specific gravity of the vehicle.
Storage conditions: At ambient temperature.
TREATMENT:
Method: Oral gavage, using a plastic feeding tube. Formulations were placed on a magnetic stirrer during dosing.
Dose volume: 10 mL/kg body weight. Actual dose volumes were calculated weekly according to the latest body weight. - Analytical verification of doses or concentrations:
- no
- Details on analytical verification of doses or concentrations:
- No analytical method could be developed to determine the test substance in formulations with propylene glycol.
No analytical method based on gas chromatography (GC), liquid chromatography (HPLC), UV spectrophotometry or total organic carbon analysis (TOC) could be developed for the determination of the test substance in formulations. - Duration of treatment / exposure:
- 13 weeks. Animals were dosed up to the day prior to necropsy.
- Frequency of treatment:
- Once daily, 7 days per week, approximately at the same time each day with a maximum of 5 hours difference between the earliest and latest dose.
Doses / concentrations
- Remarks:
- Doses / Concentrations:
0 (vehicle control), 10, 50, 250 mg/kg/day
Basis:
other: nominal ingested
- No. of animals per sex per dose:
- 10 males and 10 females per dose.
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: Based on the results of a 28-day oral study, the dose levels for the 90-day oral gavage study were selected to be 0, 10, 50 and 250 mg/kg/day.
- Positive control:
- No positive control.
Examinations
- Observations and examinations performed and frequency:
- MORTALITY/VIABILITY:
At least twice daily.
CLINICAL SIGNS:
At least once daily, detailed clinical observations were made in all animals. Once prior to start of treatment and at weekly intervals, this was also performed outside the home cage in a standard arena. The time of onset, degree and duration were recorded. All symptoms were recorded and graded according to fixed scales:
Maximum grade 1: grade 0 = absent, grade 1 = present
Maximum grade 3 or 4: grade 1 = slight, grade 2 = moderate, grade 3 = severe, grade 4 = very severe
FUNCTIONAL OBSERVATIONS:
During week 12-13 of treatment, the following tests were performed on all animals:
- hearing ability, pupillary reflex, static righting reflex and grip strength.
- motor activity test (recording period: 12 hours during overnight for individual animals, using a computerised monitoring system.
OPHTHALMOSCOPIC EXAMINATION (direct):
Following instillation of tropicamide solution (5 mg/ml), both eyes were examined by means of an ophthalmoscope at pretest (All animals) and at week 13 (Group 1 (control) and Group 4 (250 mg/kg/day).
Since no treatment related findings were noted in week 13, the eyes if the rats in Groups 2 (10 mg/kg/day) and 3 (50 mg/kg/day) were not examined
BODYWEIGHTS:
Weekly
FOOD CONSUMPTION: Weekly
WATER CONSUMPTION:
Since a treatment-related increase in water consumption was suspected based on qualitative assessment of water consumption, water consumption was determined daily from day 69 onwards, except over those days where the animals were placed individually in the motor activity device. Subjective appraisal of water intake was maintained during the remainder of the study.
CLINICAL LABORATORY INVESTIGATIONS:
HAEMATOLOGY:
Blood samples were collected from all surviving animals under iso-flurane anaesthesia immediately prior to scheduled post mortem examination. The animals were fasted overnight before blood sampling, but water was provided. Blood samples were drawn from the retro-orbital sinus and collected into tubes prepared with EDTA for haematological parameters (0.5 mL), with citrate for clotting test (0.9 mL) and Li-heparin treated tubes for clinical biochemistry parameters (0.5 mL). The following parameters were determined.
Haematology: white blood cells, differential leucocyte count (neutrophils, lymphocytes, monocytes, eosinophils, basophils), red blood cells, reticulocytes, red blood cell distribution width, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, platelets.
Clotting Potential: Prothrombin time, activated partial thromboplastin time.
Clinical Biochemistry: Alanine aminotransferase, Aspartate aminotransferase, alkaline phosphatase, total protein, albumin, total bilirubin, urea, creatinine, glucose, cholesterol, sodium, potassium, chloride, calcium, inorganic phosphate. - Sacrifice and pathology:
- PATHOLOGY:
NECROPSY:
All animals surviving to the end of the observation period were deeply anaesthetised using isoflurane vapour and subsequently exsanguinated. All animals assigned to the study were necropsied and descriptions of all macroscopic abnormalities recorded. Samples of the following tissues and organs were collected from all animals at necropsy and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution):
Identification marks: not processed, Adrenal glands, aorta, brain [cerebellum, mid-brain, cortex], caecum, cervix, (clitoral gland), colon, duodenum, epididymides, eyes with optic nerve [if detectable] and (harderian gland), female mammary gland area, (femur including joint), heart, ileum, jejunum, kidneys, (larynx), (lacrimal gland, exorbital), liver, lung infused with formalin, lymph nodes - mandibular, mesenteric, (Nasopharynx), oesophagus, ovaries, pancreas, peyers patches [jejunum, ileum] if detectable, pituitary gland, (preputial gland), prostate gland, rectum, (salivary glands - mandibular, sublingual), sciatic nerve, seminal vesicles, (skeletal muscle), (skin), spinal cord - cervical, midthoracic, lumbar, spleen, sternum with bone marrow, stomach, testes, thymus, thyroid including parathyroid [if detectable], (tongue), trachea, urinary bladder, uterus, vagina, all gross lesions.
Tissues/organs mentioned in parentheses were not examined by the pathologist, since no signs of toxicity were noted at macroscopic examination.
ORGAN WEIGHTS:
The following organ weights and terminal body weight were recorded from the surviving animals on the scheduled day of necropsy:
- Adrenal glands,
- Brain,
- Epiddidymides
- Heart
- Kidneys
- Liver
- Ovaries
- Spleen
- Testes
- Thymus
- Uterus
HISTOTECHNOLOGY:
All organ and tissue samples, as defined under Histopathology (following), were processed, embedded and cut at a thickness of 2-4 micrometers and stained with haematoxylin and eosin.
HISTOPATHOLOGY:
The following slides were examined by a pathologist.
- all tissues collected at the scheduled sacrifice from all group 1 (control) and group 4 (250 mg/kg/day) animals.
- all tissues from animal no.57 that was found dead
- all gross lesions.
All abnormalities were described. An attempt was made to correlate gross observations with microscopic findings.
Tissues/organs mentioned in parentheses (see necropsy section) were not examined by a pathologist, since no signs of toxicity were noted at macroscopic examination. - Statistics:
- The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test1 (many-to-one t-test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-to-one rank test) was applied when the data could not be assumed to follow a normal distribution.
- The exact Fisher-test was applied to frequency data.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Results and discussion
Results of examinations
- Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- no effects observed
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Description (incidence and severity):
- No toxicologically relevant changes
- Clinical biochemistry findings:
- no effects observed
- Description (incidence and severity):
- No toxicologically relevant changes
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Description (incidence and severity):
- No toxicologically significant changes
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- No toxicologically significant changes
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- No toxicologically significant changes
- Histopathological findings: neoplastic:
- not examined
- Details on results:
- MORTALITY:
No mortality occurred during the study period that was considered to be related to treatment with the test substance.
One female at 10 mg/kg/day (no. 57) was found dead after dosing on day 11 of the study. Necropsy of this animal revealed a perforated oesophagus and watery-clear fluid in the thoracic cavity. These signs were considered evidence of a misgavage.
CLINICAL SIGNS:
There were no clinical signs of toxicity noted during the observation period.
Salivation seen after dosing among all groups treated with the test substance is often noted in rats of this age and strain following oral gavage. Considering the nature and minor severity of the effect and its time of occurrence (i.e. after dosing), this was considered to be a physiological response rather than a sign of systemic toxicity.
Other incidental findings that were noted included alopecia, scabs, clonic spasms (one female at 50 mg/kg/day), and a broken tail apex. These findings are occasionally noted in rats of this age and strain which are housed and treated under the conditions in this study. At the incidence observed, these were considered signs of no toxicological significance.
FUNCTIONAL OBSERVATIONS:
Hearing ability, pupillary reflex and static righting reflex were normal in all animals. No toxicologically relevant changes in grip strength were observed.
A few animals among the dose group showed an abnormal grip strength. The incidence of this finding was unrelated to the dose. Therefore, this was considered to be of no toxicological relevance. The variation in motor activity did not indicate a relation with treatment.
OPHTHALMOSCOPIC EXAMINATION:
There were no ophthalmology findings at pre-dose and in week 13.
BODY WEIGHTS:
Body weights and body weight gain of treated animals remained in the same range as controls over the study period.
Slight weight loss was noted for most females of all dose groups in the last week of treatment. No explanation could be provided for this finding. As the incidence and degree of slight weight loss was similar between treated and control groups, it was considered that there was no relation to treatment with the test substance.
FOOD CONSUMPTION:
Food consumption before or after allowance for body weight was similar between treated and control animals.
Food consumption (before or after allowance for body weight) of all groups of males and females was lower than normally encountered for rats of this age and strain. It was considered that this was due to the volume of propylene glycol administered and subsequent lower energy demand via food intake.
WATER CONSUMPTION:
Water consumption determined from day 69 onwards was similar between treated and control animals.
CLINICAL LABORATORY INVESTIGATIONS:
HAEMATOLOGY:
No toxicologically relevant changes occurred in haematological parameters of treated rats.
The statistically significant lower haemoglobin level of females at 50 mg/kg/day occurred in the absence of a treatment-related distribution and remained within the range considered normal for rate of this age and strain. No toxicological significance was therefore ascribed to this change.
CLINICAL BIOCHEMISTRY:
No toxicologically relevant changes occurred in clinical biochemistry parameters of treated rats.
A few males among the dose groups showed high aspartate aminotransferase activity levels.The incidence of these higher values was not related to the dose. However, in several animals that showed higher aspartate aminotransferase activity levels a centrilobular vacuolation of the liver was apparent It was therefore considered that these higher individual aspartate aminotransferase activity levels were due to the volume of vehicle (propylene glycol) administered, and did not reflect toxicity of the test substance.
Any statistically significant changes observed among female dose groups were considered to be of no toxicological significance as they occurred in the absence of a treatment-related distribution. These alterations included higher glucose levels and lower sodium levels at 50 mg/kg/day, and lower calcium and inorganic phosphate levels at 10, 50 and/or 250 mg/kg/day.
PATHOLOGY:
MACROSCOPIC EXAMINATION:
Necropsy did not reveal any toxicologically relevant alterations.
An accentuated lobular pattern of the liver was noted in 1/10 control males, 3/10 males at 50 mg/kg/day and 1/10 males at 250 mg/kg/day. This finding in most cases centrilobular vacuolation of the liver and was considered to have occurred due to the volume of vehicle (propylene glycol) administered.
The large soft nodule on the kidney of one female at 10 mg/kg/day (no. 55) correlated with nephroblastoma. This was considered to be a spontaneous lesion.
Other necropsy findings were also considered to be of no toxicological significance since they occurred in the absence of a treatment-related distribution, and are occasionally seen among rats used in these types of studies. These findings included pelvic dilation of the kidneys, flaccid testes, reduced size of the testes and epididymides, a yellowish nodule on the epididymides, red foci on the mandibular lymph nodes, red discolouration of the mandibular lymph nodes, alopecia, exophthalmus, a cyst on the ovaries, fluid in the uterus, and red discolouration of the thymus.
ORGAN WEIGHTS:
No toxicologically significant changes were noted in organ weights and organ to body weight ratios.
Any statistically significant changes that were noted (i.e. lower adrenal and kidney to body weight ratios in males at 10 and 250 mg/kg/day respectively) were considered not to be a sign of toxicity as a dose-related distribution was absent and mean remained within the range considered normal for rats of this age and strain.
MICROSCOPIC EXAMINATION:
There were no microscopic findings recorded which could be attributed to treatment with the test substance.
In the liver of a few male rats vacuolation of the centrilobular area was recorded, in most cases correlating with the accentuated lobular pattern and higher liver weights recorded at necropsy. This was seen in 2/10 males of group 1 (minimal/slight), in 2/10 males of group 3 (moderate) and in 5/10 males of group 4 (two animals: minimal, three animals: slight). The vacuolation recorded in these animals is probably due to the high volume of propylene glycol (10 ml/kg/day) administered to the animals.
In animal 55 (female group 2) a nephroblastoma was recorded in the right kidney, correlating with the nodule recorded at necropsy. This finding was considered to be spontaneous in nature. All other microscopic findings were within the range of background pathology encountered in Wistar rats of this age and strain and occurred at similar incidences and severity in both control and treated rats.
Effect levels
- Dose descriptor:
- NOAEL
- Effect level:
- 250 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: No toxicologically significant changes were noted in any of the parameters investigated
Target system / organ toxicity
- Critical effects observed:
- not specified
Any other information on results incl. tables
Discussion
No toxicologically significant changes were noted in any of the parameters investigated in this study (i.e. clinical appearance, functional observations, ophthalmoscopy, body weight, food consumption, water consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).
A few cases of vacuolation of the centrilobular area of the liver were noted among male dose groups (including the control group), along with incidental cases of higher liver weights and higher aspartate aminotransferase activity levels. The incidence of these findings was limited to a few males only and was not related to the administered dose of test substance. It was considered that these findings occurred due to the high volume of vehicle (propylene glycol) administered to the animals. The volume of administered propylene glycol (i.e. 10 ml/kg) was within guideline limits. Among all other males and females, no such findings were noted. Therefore, it was considered that this did not adversely affect interpretation of the study results.
Applicant's summary and conclusion
- Conclusions:
- Wistar rats were treated with AMIDE#71 for 13 weeks by daily oral gavage at dose levels up to 250 mg/kg/day.
No toxicologically significant changes were noted in any of the parameters investigated in the study (i.e. clinical appearance, functional observations, ophthalmoscopy, body weight, food consumption, water consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).
From the results presented in this report a definitive No Observed Adverse Effect Level (NOAEL) for AMIDE #71 of 250 mg/kg/day was established. - Executive summary:
Title:
Repeated dose 90-Day oral toxicity study with AMIDE#71 by daily gavage in the rat.
Guidelines:
The study was based on the following guidelines.
- EC Directive 67/548/EEC, B Repeated Dose (90 days) Toxicity (oral), 2001
- OECD 408, Repeated Dose 90-day Oral Toxicity Study in Rodents, 1998
- OPPTS 870.3100, EPA 712-C-98 -199, 90-Day Oral Toxicity in Rodents, 1998
- Japanese Chemical Substances Control Law 1987, Notification of Nov.21 2003 by MHLW (No.1121002), METI (No.2) and ME (No.031121002)
Rationale for Dose Levels:
Based on the results of a 28-day oral study, the dose levels for the 90 -day oral gavage study were selected to be 0, 10, 50 and 250 mg/kg/day.
Study outline:
The test substance was administered daily for at least 90 days by oral gavage to Wistar rats. One control group and three treated groups were tested, each consisting of 10 males and 10 females.
Evaluated parameters:
The following parameters were evaluated: clinical signs daily; functional observation tests in week 12 -13; body weight and food consumption weekly; water consumption daily from day 69 onwards and subjective appraisal on other days; ophthalmoscopy at pretest and in week 13; clinical pathology and macroscopy at termination; organ weights and histopathology on a selection of tissues.
Results:
No analytical method could be developed to determine the test substance in formulations with propylene glycol.
No toxicologically significant changes were noted in any of the parameters investigated in the study (i.e. clinical appearance, functional observations, ophthalmoscopy, body weight, food consumption, water consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).
A few cases of vacuolation of the centrilobular area of the liver were noted among male dose groups (including the control group), along with incidental cases of higher liver weights and higher aspartate aminotransferase activity levels. The incidence of these findings was limited to a few males only and was not related to the administered dose of test substance. It was considered that these findings occurred due to the high volume of vehicle (propylene glycol) administered to the animals. The volume of administered propylene glycol (i.e. 10 ml/kg) was within guideline limits. Among all other males and females, no such findings were noted. Therefore, it was considered that this did not adversely affect interpretation of the study results.
Conclusion:
From the results presented in this report a definitive No Observed Adverse Effect Level (NOAEL) for AMIDE #71 of 250 mg/kg/day was established.
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