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EC number: 221-070-0 | CAS number: 2997-92-4
- 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://poisoncentres.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:
- 30.05.-27.09.2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 012
- Report date:
- 2012
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
- Deviations:
- yes
- Remarks:
- minor deviations in the experimental procedure which were rated uncritical (see Results and discussion)
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
Test material
- Reference substance name:
- 2,2'-azobis[2-methylpropionamidine] dihydrochloride
- EC Number:
- 221-070-0
- EC Name:
- 2,2'-azobis[2-methylpropionamidine] dihydrochloride
- Cas Number:
- 2997-92-4
- Molecular formula:
- C8H18N6.2ClH
- IUPAC Name:
- 2,2'-diazene-1,2-diylbis(2-methylpropanimidamide) dihydrochloride
- Reference substance name:
- 2,2'-azobis(2-amidinopropane)-dihydrochloride
- IUPAC Name:
- 2,2'-azobis(2-amidinopropane)-dihydrochloride
- Test material form:
- other: granules
- Details on test material:
- - Name of test material (as cited in study report): 2,2'-azobis(2-amidinopropane)-dihydrochloride
- Substance type: organic
- Physical state: solid
- Analytical purity: 99.1 %
- Lot/batch No.: I59596
- Expiration date of the lot/batch: 31. Dec. 2012
- Storage condition of test material: ambient temperature, 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, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: 7-8 weeks
- Weight at study initiation: no data
- Fasting period before study: no
- Housing: Groups of five animals of the same sex in open macroloncages type 2000P
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- water
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test item was suspended in tab-water. For the high dose group (75 mg/kg BW), 750 mg of the test item were suspended in 40 ml of sterile filtered water, yielding a concentration of 18.75 mg/m:. The preparation was intended for an spplication volume of 4 ml per kg body weight. therefore, 4 mL of this high-dose suspension contained 75 mg of the test item. For the medium-dose group (25 mg/kg bw) an aliquot of the high-dose suspension was diluted 1 in 3; it contained 6.25 mg/mL. For the low-dose group (8.3 mg/kg bw) an aliquot of the medium-dose suspension was diluted 1 in 3. The low-dose suspension contained 2.08 mg/mL
VEHICLE
- Amount of vehicle (if gavage): 4 mL per kg body weight - Frequency of treatment:
- daily
Doses / concentrationsopen allclose all
- Remarks:
- Doses / Concentrations:
75 mg/kg bw
Basis:
actual ingested
- Remarks:
- Doses / Concentrations:
25 mg/kg bw
Basis:
actual ingested
- Remarks:
- Doses / Concentrations:
8.3 mg/kg bw
Basis:
actual ingested
- No. of animals per sex per dose:
- 5
- Control animals:
- yes
- Details on study design:
- - Dose selection rationale: based on dose-range finding study
- Rationale for animal assignment (if not random): random - Positive control:
- non
Examinations
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly,
- IRWIN test
BODY WEIGHT: Yes
- Time schedule for examinations: weekly
FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: twice weekly
HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately before sacrificing
- Animals fasted: No
- How many animals: all
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately before sacrificing
- Animals fasted: No
- How many animals: all
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: last exposure week
- Dose groups that were examined: all dose groups
- Battery of functions tested: sensory activity (beam walking test) / grip strength - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes (see table)
HISTOPATHOLOGY: Yes (see table) - Statistics:
- Descriptive statistics
The data for body weight, organ weight, grip strength and haematological/biochemical
examination were documented for each individual animal. Food- and water consumption
were documented in gender groups. For each experimental group, means and standard
deviations were calculated. The test item groups were analysed in comparison to the vehicle
group by calculation of statistical significance using a two-tailed unpaired student´s t-test
(p ≤ 0,05). Statistic significances were represented when appropriate in graphical form or in
the text of the study report.
Additionally, for the haematological/biochemical data, the median, the 0,25 quantil and the
difference of median and mean were calculated in %. These calculations give basic
information about the sample distribution of the respective data sets in relation to a Gaussian
(normal) distribution. In case of differences higher or lower than 10% the Study Director
decided whether statistical analysis was still appropriate.
The main decision-tree, documented in the appendix, displays the general approach of the
deductive statistical analysis.
Most statistical hypotheses of this study are best characterised as “many to one”
comparisons, for example: a vehicle control vs. three treatment groups. Therefore, the
adequate analytic method was a One-Way ANOVA (ANAlysis Of VAriance), followed by a
post hoc t-test.
For interval-scaled data, it was intended to apply a One-Way ANOVA supplemented by
Dunnett’s post-hoc t-test. Ordinal-scaled data should be analysed using the Kruskal-Wallis
test, supplemented by Dunn’s t-test.
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:
- not examined
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- see Details
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- see Details
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- see Details
- Gross pathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Histopathological findings: neoplastic:
- no effects observed
- Details on results:
- Analyses of haematology and serum biochemistry returened normal values in the vast majority of the monitored parameters. The minor effects observed in individual red blood cells related parameters, the thrombocytes as well as the total number of leucocytes were assessed to be of no toxicological relevance.
A test item related effect on the potassium concentration in the blood of male animals of the high dose group was found to be associated with increased kidney weight and decreased size of the adrenal gland.
Effect levels
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- ca. 25 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: A decrease of Potassium concentration in the blood along with increased kidney weight and decreased size of the adrenal gland.
Target system / organ toxicity
- Critical effects observed:
- not specified
Applicant's summary and conclusion
- Conclusions:
- A NOAEL of 25 mg/kg bw/ day for the 28-day repeated dose toxicity has been established.
- Executive summary:
Aim of the study:
The aim of this study was to assess data on the subacute toxicity of the chemical substance 2,2' – azobis (2-amidinopropane)-dihydrochloride suspended in water.
Experimental model:
The test item 2,2' – azobis (2-amidinopropane)-dihydrochloride was administered daily by oral gavage at dose levels of 75, 25 and 8 mg/kg bodyweight per day to 5 male and 5 female Wistar rats per dose group over a period of 28 days. Additionaly, 10 male and 10 female rats received the same volume of sterile water as vehicle control. All solutions were administered at an application volume of 4 mL/kg bodyweight.
Assessed parameters:
During the in-life phase, viability, general and detailed clinical signs, food and water consumption, body weight, were recorded.
At the end of the in-life phase, grip strength and reactivity to sensory stimuli (beam walking test) were detected and blood samples from all animals were collected to provide data on haematology and serum biochemistry. All animals were sacrificed immediately after bleeding and examined by gross necropsy. Weights of selected organs were recorded and tissues and organs were preserved according to the study plan. All samples were processed for histopathological examination. The examination was conducted on samples from the high dose groups and the vehicle groups.
Assessment of the Results:
Detailed clinical signs, food and water consumption, motoric activity, and reactivity to sensory stimuli showed no distinct abnormalities. The grip strength test also revealed no differences between the different experimental groups.
The mean body weight increase of all experimental groups was within the normal range for rats of this strain and age.
Analyses of haematology and serum biochemistry returned normal values in the vast majority of the monitored parameters. The minor effects observed in individual red blood cells related parameters, the thrombocytes as well as the total number of leucocytes were assessed to be of no toxicological relevance. A test item related effect on the potassium concentration in the blood of male animals of the high dose group was found to be associated with increased kidney weight and decreased size of the adrenal gland.
Morphological and histological examination did not reveal morphological changes related to the test item. Macroscopic findings in all animals as well as histopathological findings in animals of the high dose group did not differ significantly or the differences were regarded as coincidental. All findings are considered to be spontaneous in nature and within the normal background pathology commonly seen in rats of this strain and age.
Conclusion:
A daily oral administration of the test item, 2,2'-azobis(2-amidinopropane)-dihydrochloride, to Wistar rats at a dose level of 75, 25 and 8 mg/kg body weight over a time period of 28 days resulted in minor systemic effects in the male high dose group. Concerning the findings in kidney weight, adrenal-gland size and potassium levels in the male high dose group, the NOAEL in this study was 25 mg/kg body weight over a 28 day application period.
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