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EC number: 220-666-8 | CAS number: 2855-13-2
- 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
Genetic toxicity: in vivo
Administrative data
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1990-04-17 - 1990-05-17
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 990
- Report date:
- 1990
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Version / remarks:
- 1983
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
- Version / remarks:
- 1984
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
Test material
- Reference substance name:
- 3-aminomethyl-3,5,5-trimethylcyclohexylamine
- EC Number:
- 220-666-8
- EC Name:
- 3-aminomethyl-3,5,5-trimethylcyclohexylamine
- Cas Number:
- 2855-13-2
- Molecular formula:
- C10H22N2
- IUPAC Name:
- 3-aminomethyl-3,5,5-trimethylcyclohexylamine
- Reference substance name:
- Isophorone diamine
- IUPAC Name:
- Isophorone diamine
- Details on test material:
- Isophorone diamine of Hüls AG, produced 20 March 1990, ID No. 3641/81172; purity 99.85 % (GC)
Constituent 1
Constituent 2
Test animals
- Species:
- mouse
- Strain:
- NMRI
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ORGANISMS:
- Supplier: BRL Tierfarm Füllinsdorf (Switzerland)
- Age: minimum 10 weeks + 5 days acclimatization
- Weight at study initiation: approximately 30 g
- Fasting period before study: 18 hours
- Housing: single, Makrolon Type I, with wire mesh top
- Diet: ALTROMIN standard diet, ad libitum
- Water: tap water ad libitum
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 3 °C
- Humidity (%): 30 - 70 %
- Photoperiod (hrs dark / hrs light): 12h / 12h
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- aqua dest.
- Details on exposure:
- PRE-EXPERIMENT FOR TOXICITY
500 mg/kg bw was estimated to be the maximum tolerated dose, cytotoxic reactions were not observed
TREATMENT
Approximately 18 hours before treatment with the test article the animals received no food but water ad libitum. At the beginning of the treatment the animals were weighed and the individual volume to be administered was adjusted to the animals body weight. The animals received the test article once. Twelve animals, six males and six females, were treated per dose group. Sampling of the bone marrow was done 24, 48 and 72 hours after treatment.
doses: 0, 50, 150, 500 mg/ kg b.w.
positive control: cyclophosphamide, dissolved in physiol. saline, 40 mg/kg bw
negative control: vehicle - Duration of treatment / exposure:
- single dose
- Frequency of treatment:
- single dose
- Post exposure period:
- 24, 48 or 72 hours
Doses / concentrations
- Remarks:
- Doses / Concentrations:
50, 150, or 500 mg/kg, dissolved in 10 ml/kg bw dose volume
Basis:
actual ingested
- No. of animals per sex per dose:
- 6 per dosage group and sex with 3 cases of post-treatment duration for negative control and treated groups totals 4 x 3 + 1 groups x 6 animals x 2 sexes = 156 animals; only 5 animals per group and sex were evaluated
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- cyclophosphamide, dissolved in physiol. saline, 40 mg/kg bw
Examinations
- Tissues and cell types examined:
- femora, bone marrow
- Details of tissue and slide preparation:
- PREPARATION OF THE ANIMALS::
The animals were sacrificed by cervical dislocation. The femora were removed, the epiphyses were cut off and the marrow was flushed out with fetal calf serum, using a 5 ml syringe. The cell suspension was centrifuged at 1,500 rpm for 5 minutes and the supernatant was discarded. A small drop of the resuspended cell pellet was spread on a slide. The smear was air-dried and then stained with May-Grilnwald (MERCK, D-6100 Darmstadt, F.R.G.) /Giemsa (Gurr, BDH Limited Poole, Great Britain). Cover slips were mounted with EUKITT (KINDLER, D-7800 Freiburg F.R.G.). At least one slide was made from each bone marrow sample.
ANALYSIS OF CELLS:
Evaluation of the slides was performed using NIKON microscopes with 100x oil immersion objectives. 1000 polychromatic erythrocytes (PCE) were analysed per animal for micronuclei. To describe a cytotoxic effect the ratio between polychromatic and normochromatic erythrocytes was determined in the same sample and expressed as normochromatic erythrocytes per 1000 PCEs. The analysis was performed with coded slides. Five animals per sex and group were evaluated as described. The remaining animal of each test group was evaluated in case an animal had died in its test group spontaneously or due to gavage error. - Evaluation criteria:
- A test article is classified as mutagenic if it induces either a statistically significant dose-related increase in the number of micronucleated polychromatic erythrocytes or a reproducible statistically significant positive response for at least one of the test points. A test article producing neither a statistically significant dose-related increase in the number of micronucleated polychromatic erythrocytes nor a statistically significant and reproducible positive response at anyone of the test points is considered nonmutagenic in this system. This can be confirmed by means of the nonparametric Mann-Whitney test. However, both biological and statistical significance should be
considered together. - Statistics:
- Statistical significance at the five per cent level (p < 0. 05) was evaluated by means of the non-parametric Mann-Whitney test.
Results and discussion
Test results
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- Maximum tolerated dose determined in pre-experiment
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- EFFECT ON PCE/NCE RATIO: not affected (PCE = polychromatic erythrocytes, NCE = normochromatic erythrocytes)
GENOTOXIC EFFECTS:
In comparison to the corresponding negative controls there was no enhancement in the frequency of the detected micronuclei at preparation intervals 48 hours and 72 hours. The mean values of micronuclei observed after treatment with Isophorondiamine (50, 150, and 500 mg/kg b.w.) were in the same range as compared to the corresponding negative control groups. At preparation interval 24 hours a statistically significant increase of the micronucleus frequency after administration of 500 mg/kg b.w. Isophorondiamine was proved by biometric analysis ( p< 0.05, Mann-Whitney test).
This statistical significance is considered to be of minor importance:
1. The corresponding actual negative control rate in this study was very low (0.02%). The mean historical negative control value obtained within the last 12 months was 0.073 %. The range was 0.04% - 0.12%.
2. The frequency of 0.10 % PCEs with micronuclei after treatment with 500 mg/kg b.w. Isophorondiamine is within the range of the historical control data presented.
3. The value of 0.10 deviates not substantially from the obtained at preparation intervals 48 h (0.06 %) and (0.09 %).
Therefore, the statistical significance is not considered to be an indication for an induced mutagenic effect due to the test article.
40 mg/kg b.w. cyclophosphamide administered per os was used as positive control which showed a distinct increase in induced micronucleus frequency.
Any other information on results incl. tables
-------------------------------------------------------- Treatment Time % Micron. in PCE PCE/NCE -------------------------------------------------------- Vehicle 24 h 0.02 (0 - 2) 1000 / 970 50 mg TS/kg bw 24 h 0.07 (0 - 2) 1000 / 917 150 mg TS/kg bw 24 h 0.06 (0 - 2) 1000 / 892 500 mg TS/kg bw 24 h 0.10 (0 - 3) * 1000 / 908 40 mg CPA/kg bw 24 h 1.08 (2 -22) * 1000 /1127 Vehicle 48 h 0.04 (0 - 1) 1000 / 699 50 mg TS/kg bw 48 h 0.06 (0 - 2) 1000 / 676 150 mg TS/kg bw 48 h 0.06 (0 - 3) 1000 / 702 500 mg TS/kg bw 48 h 0.09 (0 - 3) 1000 / 781 Vehicle 72 h 0.14 (0 - 5) 1000 / 744 50 mg TS/kg bw 72 h 0.07 (0 - 3) 1000 / 681 150 mg TS/kg bw 72 h 0.13 (0 - 4) 1000 / 754 500 mg TS/kg bw 72 h 0.16 (0 - 4) 1000 / 758 -------------------------------------------------------- TS = test substance; CPA = cyclophosphamide; * p<0.05
Applicant's summary and conclusion
- Conclusions:
- It can be stated that during the study described and under the experimental conditions reported, the test article did not induce micronuclei as determined by the micronucleus test with bone marrow cells of the mouse.
- Executive summary:
The test articlewas assessed in the micronucleus assay for its potential to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse.
The test article was dissolved in aqua dest .. This solvent was used as negative control. The volume administered orally was 10 ml/kg body weight (b.w.). 24 h, 48 h and 72 h after a single application of the test article the bone marrow cells were
collected for micronuclei analysis. Ten animals (5 males, 5 females) per test group were evaluated for the occurrence of micronuclei in PCEs. Per animal 1000 PCEs
were scored for micronuclei. To describe a cytotoxic effect due to the treatment with the test article the ratio between polychromatic and normochromatic erythrocytes (NCE) was determined in the same sample and reported as number of NCEs per 1000 PCEs.
The following dose levels of the test article were investigated:
24 h preparation interval: 50, 150 and 500 mg/kg b.w.
48 h preparation interval: 50, 150 and 500 mg/kg b.w.
72 h preparation interval: 50, 150 and 500 mg/kg b.w.
In pre-experiments 500 mg/kg b.w. was estimated to be the maximum tolerated dose. The animals expressed toxic reactions.
The ratio of normochromatic to polychromatic erythrocytes was not affected by the treatment with Isophorondiamine, indicating that the test article had no cytotoxic properties at the dose levels tested.In conclusion, it can be stated that during the study described and under the experimental conditions reported, the test article did not induce micronuclei as determined by the micronucleus test with bone marrow cells of the mouse.
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