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EC number: 234-147-9 | CAS number: 10563-26-5
- 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
Oral: NOAEL = 30 mg/kg bw/day (OECD 422, GLP, BASF 2013)
Key value for chemical safety assessment
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- short-term repeated dose toxicity: oral
- Remarks:
- combined repeated dose and reproduction / developmental screening
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP Guideline Study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- 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: approx. 11 weeks
- Weight at study initiation: mean: females: 201 g, males: 318 g
- Housing: in groups of 5 animals/sex/cage
- Diet: free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water: ad libitum
- Acclimation period: at least five days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 40 - 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- oral: gavage
- Vehicle:
- water
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: Formulations (w/w) were prepared daily within 5 hours prior to dosing and were homogenized to a visually acceptable level. Adjustment was made for specific gravity of the test substance. No correction was made for the purity of the test substance.
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- 1H-NMR spectroscopy
- Duration of treatment / exposure:
- females: 42-53 days
males: 29 days - Frequency of treatment:
- daily
- Remarks:
- Doses / Concentrations:
30, 100, 300 mg/kg bw/day
Basis:
actual ingested - No. of animals per sex per dose:
- 10
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: Based on results of the 14-day dose range finding study (Project 499971; BASF Project 01R0402/04X033) where dose levels of 0, 150 and 400 mg/kg bw/day were assessed. Animals at 400 mg/kg bw/day had slightly lower body weight gains or weight loss and slightly lower food consumption. Changes in haematology and clinical biochemistry parameters were noted, along with increased absolute and relative liver weights (both sexes) and higher relative kidney weights (males). Females at 150 mg/kg bw/day also had higher absolute and relative liver weights.
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations checked: mortality, pain, distress or discomfort
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on Days 1 and 4.
FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
WATER CONSUMPTION AND COMPOUND INTAKE : Yes: Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no treatment related effect was suspected.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: on day of necropsy
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 5 animals/sex/group
- Parameters examined: white blood cells (WBC), differential leucocyte count (neutrophils, lymphocytes, monocytes, eosinophils, basophils), red blood cells, reticulocytes, red blood cell distribution width, haemoglobin, haematocrit, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), platelets, prothrombin time (PT), activated partial thromboplastin time (APTT)
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on day of necropsy
- Animals fasted: Yes
- How many animals: 5 animals/sex/group
- Parameters examined: alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), alkaline phosphatase (ALP), total protein, albumin, total bilirubin, urea, creatinine, glucose, cholesterol, sodium, potassium, chloride, calcium, inorganic phosphate, bile acids
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: daily
- Dose groups that were examined: 5 animals/sex/group
- Battery of functions tested: hearing ability, pupillary reflex, static righting reflex, grip stenght, locomotor activity - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes (macroscopic examination of the cranial, thoracic and abdominal tissues and organs, with special attention being paid to the reproductive organs)
HISTOPATHOLOGY: Yes (Adrenal glands, Ovaries, (Pancreas), (Aorta), Peyer's patches [jejunum, ileum] if detectable, Brain - cerebellum, mid-brain, cortex, Pituitary gland, Caecum, Preputial gland, Cervix, Prostate gland, Clitoral gland, Rectum, Colon, (Salivary glands - mandibular, sublingual), Coagulation gland, Sciatic nerve, Duodenum, Seminal vesicles, Epididymides, Skeletal muscle, Eyes (with optic nerve (if detectable) and Harderian gland), (Skin), Spinal cord -cervical, midthoracic, lumbar, (Male and Female mammary gland area), Spleen, Femur including joint, Sternum with bone marrow, Heart, Stomach, Ileum, Testes, Jejunum, Thymus, Kidneys, Thyroid including parathyroid if detectable, (Lacrimal gland, exorbital), (Tongue), (Larynx), Trachea, Liver, Urinary bladder, Lung, infused with formalin, Uterus, Lymph nodes - mandibular, mesenteric, Vagina
[Tissues/organs mentioned in parentheses were not examined by the pathologist, since no signs of toxicity were noted at macroscopic examination.] - Other examinations:
- Organ weights: Adrenal glands, Spleen, Brain, Testes, Epididymides, Thymus, Heart, Uterus (including cervix), Kidneys, Prostate, Liver, Seminal vesicles including coagulating glands, Ovaries, Thyroid including parathyroid
- Statistics:
- The following statistical methods were used to analyze the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test 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 if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test was applied to frequency data.
The following additional methods of statistical analysis were used:
Motor activity data was subjected to the Kruskal-Wallis nonparametric ANOVA test to determine intergroup differences followed by the Wilcoxon test to compare the treated groups to the control group.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance. - Details on results:
- CLINICAL SIGNS AND MORTALITY: At 300 mg/kg bw/day there were three males that died spontaneously (nos. 31, 32 and 38) on study days 21, 25, and 29 and one that died after blood collection immediately prior to the scheduled necropsy (no. 37), and there was one female (no. 72) that was euthanized in extremis on day 7 post coitum.
Clinical signs noted for the animals that died prematurely or were euthanized in extremis included lethargy, hunched posture, piloerection and quick breathing. These were noted on single or limited occasions on the day(s) immediately preceding death of these animals. Alopecia was noted for a single female at 300 and chromodacryorrhea (of the right periorbital region) was recorded for a single male at 100 mg/kg bw/day. These were the only other clinical signs observed and were incidental in nature.
BODY WEIGHT AND WEIGHT GAIN: Body weights and body weight gains were significantly lower than controls for males at 300 mg/kg bw/day during the mating period. Males slightly lost weight during the second week of the study and during mating period. Body weights and/or body weight gains were significantly lower during the post coitum and lactation periods for females at 300 mg/kg bw/day. Absolute body weights were lower during the post coitum and lactation periods for females at 100 mg/kg bw/day as well, though the differences were not always statistically significant. Body weight gain was significantly lower for females at 100 mg/kg bw/day on Day 8 of the pre mating period. In the absence of a dose response effect it was not considered toxicologically relevant.
FOOD CONSUMPTION AND COMPOUND INTAKE: Absolute food consumption was lower during the mating period, and relative food consumption was lower during the last part of the premating period and throughout the mating period for males at 300 mg/kg bw/day. Females at this dose level had significantly lower absolute food consumption during days 0-7 and 14-20 of the post coitum period and over days 1-4 of the lactation period. There were no other treatment related effects on absolute or relative food consumption up to 100 mg/kg bw/day.
HAEMATOLOGY: At 300 mg/kg bw/day the following statistically significant changes distinguished treated animals from controls:
-Increased white blood cells (WBC; males only)
-Increased neutrophils (males only)
-Increased monocytes (also at 100 mg/kg bw/day, males only)
-Increased reticulocytes (males only)
-Increased platelets (males only)
-Reduced lymphocytes (males only)
-Reduced mean corpuscular volume (MCV; females only)
-Reduced mean corpuscular haemoglobin (MCH; females only)
In the absence of any other findings, the higher monocytes counts at 100 mg/kg bw/day are considered not to be toxicologically relevant.
There were no other differences between treated and control animals.
CLINICAL CHEMISTRY: At 300 mg/kg bw/day, the following statistically significant changes in clinical biochemistry parameters distinguished treated animals from control animals:
-Reduced alanine aminotransferase (ALAT, females only)
-Reduced total protein (males only)
-Reduced albumin (males only)
-Increased cholesterol (males only)
-Increased bile acids (males only)
-Reduced potassium (males only)
-Reduced chloride (males only)
-Reduced calcium (males and females, also reduced for females at 100 mg/kg bw/day)
In the absence of any other findings, the lower calcium concentrations at 100 mg/kg bw/day are considered not to be toxicologically relevant.
Potassium was significantly increased for males at 100 mg/kg bw/day. This was not considered to be treatment related as no dose response effect was seen (potassium was significantly decreased for males at 300 mg/kg bw/day). Similarly, the statistically significant reduction in inorganic phosphate noted for males at 30 mg/kg bw/day was not considered to be treatment related as it occurred in the absence of a dose response relationship.
ORGAN WEIGHTS: Terminal body weights were significantly lower for animals of both sexes at 300 mg/kg bw/day (not statistically significant for females). Males at this dose level also had significantly higher absolute and relative spleen weights (relative spleen weights were also higher for males at 100 mg/kg bw/day), higher relative liver and kidney weights and significantly lower absolute testes and seminal vesicle weights. Females at this dose level had significantly higher thymus (absolute and relative) and significantly lower thyroid (absolute and relative) and adrenal (absolute) weights. The significantly increased brain to body weight ratio seen for males at 300 mg/kg bw/day was secondary to the lower terminal body weights and was not considered to be toxicologically relevant.
Similarly, the significantly higher relative thyroid weights that were noted for females at 30 mg/kg bw/day occurred in the absence of a dose response relationship and were not considered to be toxicologically relevant.
Other organ weights and organ to body weight ratios were similar between control and treated groups.
GROSS PATHOLOGY: Macroscopic findings noted for the animals that died spontaneously or were killed in extremis included beginning or advanced autolysis, dark red, reddish, or grey-white discoloration of the lungs, many dark red foci on the lungs, dark red contents of the small intestines, irregular surface of, and a tan and/or reddish focus on the stomach or forestomach, reddish foci on the stomach glandular mucosa, stomach limiting ridge thickened, greenish foci on the caecum, reddish discoloration of the thymus, black and/or dark red discoloration of the mesenteric or mandibular lymph node, agenesis and/or reduced size of the seminal vesicles.
Relevant macroscopic findings noted for animals at 300 mg/kg bw/day that survived to the scheduled necropsy included many dark red foci or an isolated tan focus on the the lungs, reddish foci on the stomach glandular mucosa and irregular surface of the forestomach. Additionally, female no. 73 was noted with a dark red and hard nodule in the right uterine horn. Incidental findings noted for control and treated animals included reddish or dark red foci on the thymus, reddish discoloration of the thymus, yellowish focus on the right medial lobe of the liver, alopecia, yellowish or greenish soft nodules on the epididymides, pelvic dilation of the kidney(s), watery-clear cyst on the right kidney, tan discoloration of the right clitoral gland, and uterus contains fluid. These findings occurred without a treatment related distribution and remained within the background range of findings seen for rats of this age and strain, and were thus not considered to be treatment related or toxicologically relevant.
HISTOPATHOLOGY: Treatment-related microscopic findings were seen in several organs including:
Lung:
- Hemorrhage was noted in 4/8 males and 2/6 females (up to moderate) treated at 300 mg/kg bw/day.
- Mucous cell hyperplasia of the bronchi/bronchioles epithelium was noted in 6/8 males (up to moderate) and in 2/6 females (up to marked) treated at 300 mg/kg bw/day.
- Bronchiolization of the alveoli wall was noted in 6/8 males (up to moderate) and in 3/6 females (up to moderate) treated at 300 mg/kg bw/day.
- Alveolar inflammation was noted at increased incidence and severity in 7/8 males (up to moderate) and 5/6 females (up to marked) treated at 300 mg/kg bw/day, compared to 2/5 males (minimal) in the 0 mg/kg bw/day, 2/5 males and 4/5 females (minimal) in the 30 mg/kg bw/day and 4/5 males and 2/5 females (minimal) in the 100 mg/kg bw/day treated rats.
- Interstitial inflammation was noted at increased incidence and severity in 8/8 males (up to moderate) and 4/6 females (up to moderate) treated at 300 mg/kg bw/day, compared to 1/5 males and 1/5 females (minimal) in the 30 mg/kg bw/day and 2/5 males (minimal) in the 100 mg/kg bw/day treated rats.
The mixed inflammatory infiltrate consisted of lymphocytes, granulocytes and macrophages.
- Fibrin deposition was noted in 3/8 males (up to moderate) treated at 300 mg/kg bw/day.
- Fibrosis was noted in 6/8 males (up to moderate) and in 3/6 females (up to slight) treated at 300 mg/kg bw/day.
Stomach:
- Ulceration of the forestomach was noted in 4/8 males (up to marked) and 2/6 females (up to moderate) in the 300 mg/kg bw/day treated rats.
- Hyperplasia of the forestomach was noted in 7/8 males and 3/6 females (up to moderate) treated at 300 mg/kg bw/day.
- Lymphogranulocytic inflammation of the forestomach was noted in 5/8 males and 3/6 females (up to moderate) treated at 300 mg/kg bw/day.
- Lymphogranulocytic inflammation of the limiting ridge was noted at increased incidence and/or severity in 6/8 males (up to slight) and 4/6 females (minimal) at 300 mg/kg bw/day, compared to minimal degree in 1/5 males treated at 0 mg/kg bw/day, 2/5 males treated at 30 mg/kg bw/day and in 1/5 females in the 100 mg/kg bw/day group.
- Vacuolation of the limiting ridge was noted at increased incidence and/or severity in 3/8 males (up to moderate) and 4/6 females (up to slight) treated at 300 mg/kg bw/day compared to 3/5 males at minimal degree in the 100 mg/kg bw/day.
- Lymphogranulocytic inflammation of the glandular stomach was noted at increased severity in 4/8 males and 2/6 females (up to moderate) treated at 300 mg/kg bw/day, compared to 2/5 males with a minimal degree in the 100 mg/kg bw/day treated males.
- Increased apoptosis in the mucosa of the glandular stomach was noted in 1/6 females (moderate) treated at 300 mg/kg bw/day.
- Hemorrhage of the glandular mucosa was noted at slight degree in 1/8 male treated at 300 mg/kg bw/day.
Thymus:
- Lymphoid atrophy was noted at increased severity (moderate) in 1/7 females treated at 300 mg/kg bw/day compared to minimal in 1/5 females treated at 100 mg/kg bw/day. This increased severity was considered to be secondary due to the poor condition of this unscheduled death.
Spleen:
- An increase in severity of hemopoietic foci (3 minimal, 3 slight, 2 moderate) was recorded in males treated at 300 mg/kg bw/day, compared to 5/5 minimal treated at 0 mg/kg bw/day, 5/5 minimal treated at 30 mg/kg bw/day and 2/5 minimal and 3/5 slight treated at 100 mg/kg bw/day.
Kidney:
- Corticomedullary tubular basophilia was noted at increased incidence and severity in 6/6 males (slight to marked) and 4/6 females (up to slight) treated at 300 mg/kg bw/day, compared to minimal degree in 1/5 males and 1/5 females treated at 0 mg/kg bw/day, 2/6 males and 1/5 females treated at 30 mg/kg bw/day and 2/7 males and 1/5 females treated at 100 mg/kg bw/day.
- Granular cast(s)/degeneration of tubular epithelium was noted in 5/6 males (up to slight) and 1/6 females (minimal, unscheduled death) treated at 300 mg/kg bw/day.
- Tubular dilatation was noted in 5/6 males (up to slight) and 5/6 females (minimal) treated at 300 mg/kg bw/day.
An increase in incidence of hyaline cast(s) was noted in 4/6 males (minimal degree) treated at 300 mg/kg bw/day, compared to 1/6 males treated at 30 mg/kg bw/day.
Adrenal gland:
- Lymphocytic inflammation of the distal part of the zona fasciculate and zona reticularis was noted in 3/5 females (all minimal) treated at 100 mg/kg bw/day and in 1/8 males (slight) and 5/6 females (slight to marked) treated at 300 mg/kg bw/day.
Eyes:
- Retinal dysplasia (rosettes) was recorded in 5/7 males (up to slight) and 5/6 females (up to moderate) treated at 300 mg/kg bw/day, compared to 1/5 females (minimal, only unilateral) treated at 100 mg/kg bw/day.
The remainder of microscopic findings recorded were within the normal range of background pathology encountered in Wistar (Han) rats of this age.
NEUROBEHAVIOUR: Functional observations: Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all animals.
Total counts were significantly lower for males at 300 mg/kg bw/day, and ambulations were also reduced (not statistically significant); a relationship to treatment cannot be excluded. All groups showed a similar habituation profile with high activity in the first interval that decreased over the duration of the test period. - Dose descriptor:
- NOAEL
- Effect level:
- 30 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- clinical signs
- Critical effects observed:
- not specified
Reference
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- 30 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- OECD 422 - screening study
Repeated dose toxicity: inhalation - systemic effects
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
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Oral:
A reliable combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test of N4 -Amine in rats by oral gavage (GLP, OECD 422) is available (BASF 2013). The test substance was administered by daily oral gavage to male
and female Wistar Han rats at dose levels of 30, 100 and 300 mg/kg bw/day. Males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 29 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation (for 42-53 days). At 300 mg/kg bw/day there were 5 unscheduled deaths and toxicologically relevant effects on body weights, food consumption, clinical pathology, macroscopic findings, organ weight changes and microscopic findings in various organs including the stomach, lungs, spleen, kidneys, adrenal glands and eyes. Higher relative spleen weights (males) and lower body weights and microscopic findings in the adrenal glands were noted for females at 100 mg/kg bw/day. In conclusion, treatment with the test substance by oral gavage in male and female Wistar Han rats at dose levels of 30, 100 and 300 mg/kg body weight/day revealed
parental toxicity at 100 and 300 mg/kg bw/day. Based on these results, a parental NOAEL of 30 mg/kg bw/day was established. Since the highest dose induced death and severe suffering the maxium tolerated dose was exceeded. The effects found at 100 mg/kg bw/day do not show functional impairment which would justify a classification for Specific Target Organ Toxicity after repeated exposure.
Justification for selection of repeated dose toxicity via oral
route - systemic effects endpoint:
Only one study available.
Justification for classification or non-classification
Dangerous Substance Directive (67/548/EEC)
The available study is considered reliable and suitable for classification purposes under 67/548/EEC. As a result the substance is not considered to be classified for repeated dose toxicity (oral) under Directive 67/548/EEC, as amended for the 28th time in Directive 2001/59/EC.
Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for repeated dose toxicity (oral) under Regulation (EC) No. 1272/2008.
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