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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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.
EC number: 422-210-5 | CAS number: 68957-94-8 T3P
- 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
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1995-10-16 to 1995-12-13
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 996
- Report date:
- 1996
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:
- EEC Directive 92/69, L 383 A, Annex B. 12., p. 154 -156
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
Test material
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: PPA 4/94
- Expiration date of the lot/batch: June 1996
- Purity test date: March 6th, 1995
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: darkness at approximately 20°C in a fume cupboard
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: stability in the vehicle is guaranteed for 4 hours
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: On the day of the experiment the test substance was dissolved in DMSO (dried) at an appropriate concentration. A magnetic stirrer was used to keep the preparation homogeneous until dosing had been completed.
Test animals
- Species:
- mouse
- Strain:
- NMRI
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Tierzucht Schonwalde GmbH i.G., Hauptstrafte 62, 16352 Schonwalde, Germany
- Age at study initiation: approximately 8 weeks
- Weight at study initiation: males: mean 40.2 g+/- 2.73 g, females: mean 31.9 g +/- 2.93 g
- Assigned to test groups randomly: yes
- Housing: in fully air-conditioned rooms in makrolon cages type Type 3 (five animals per cage) on soft wood granulate
- Diet: rat/mice diet ssniff® R/M-H (V 1534), ad libitum ssniff® GmbH, Postbox 2039, 59480 Soest, Germany
- Water: tap water in plastic bottles, ad libitum
- Acclimation period: 1 week
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ±3°C
- Humidity (%): 50 ± 20 %
- Photoperiod (hrs dark / hrs light): 12/12
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle/solvent used: DMSO, dried
- Concentration of test material in vehicle: 0 (vehicle control) and 12.5 % (w/v)
- Amount of vehicle: total volume 10 mL/kg bw - Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
On the day of the experiment the test substance was dissolved in DMSO (dried) at an appropriate concentration. A magnetic stirrer was used to keep the preparation homogeneous until dosing had been completed. DMSO (dried) was administered in the same way to the negative control groups. The study included a simultaneous positive control using Endoxan®, which was administered once orally at a dose of 50 mg per kg body weight. - Duration of treatment / exposure:
- In conformity with the test procedure the animals were killed by carbon dioxide asphyxiation 12, 24 or 48 hours after administration.
- Frequency of treatment:
- single administration
- Post exposure period:
- 12, 24 or 48 hours
Doses / concentrationsopen allclose all
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- vehicle control, killing time 24 h p.a.
- Dose / conc.:
- 1 250 mg/kg bw/day (actual dose received)
- Remarks:
- test group, killing time 24 h p.a.
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- vehicle control, killing time 48 h
- Dose / conc.:
- 1 250 mg/kg bw/day (actual dose received)
- Remarks:
- test group, killing time 48 h
- Dose / conc.:
- 0 mg/kg bw/day (actual dose received)
- Remarks:
- vehicle control, killing time 12 h
- Dose / conc.:
- 1 250 mg/kg bw/day (actual dose received)
- Remarks:
- test group, killing time 12 h
- No. of animals per sex per dose:
- 5 male anf 5 female animals per group per killing time
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Endoxan containing cyclophosphamide
- Route of administration: oral by gavage
- Doses / concentrations: 50 mg/kg bw, killing time 24 h p.a.
Examinations
- Tissues and cell types examined:
- bone marrow erythrocytes
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION:
Oral administration of 1500 mg per kg body weight resulted in mortality in male mice. The highest sub lethal dose of 1250 mg per kg body weight was selected for the main study.
TREATMENT AND SAMPLING TIMES
In conformity with the test procedure the animals were killed by carbon dioxide asphyxiation 12, 24 or 48 hours after administration.
DETAILS OF SLIDE PREPARATION:
For each animal, about 3 mL foetal bovine serum was poured into a centrifuge tube. Both femora were removed and the bones freed of muscle tissue. The proximal ends of the femora were opened and the bone marrow flushed into the centrifuge tube. A suspension was formed. The mixture was then centrifuged for 5 minutes at approx. 1200 rpm, after which almost all the supernatant was discarded. One drop of the thoroughly mixed sediment was smeared onto a cleaned slide, identified by project code and animal number and air-dried for about 12 hours.
Staining was performed as follows:
- 5 minutes in methanol
- 5 minutes in May-Grunwald's solution
- brief rinsing twice in distilled water
- 10 minutes staining in 1 part Giemsa solution to 6 parts buffer solution, pH 7.2 (Weise)
- rinsing in distilled water
- drying
- coating with Entellan
METHOD OF ANALYSIS:
1000 polychromatic erythrocytes were counted for each animal. The number of cells with micronuclei was recorded, not the number of individual micronuclei. As a control measure 1000 mature erythrocytes were also counted and examined for micronuclei. In addition, the ratio of polychromatic to normochromatic erythrocytes was determined. All bone marrow smears for evaluation were coded to ensure that the group from which they were taken remained unknown to the investigator. The number of polychromatic erythrocytes with micronuclei occurring in the 1000 polychromatic erythrocytes, and the number of normocytes with micronuclei occurring in the 1000 normocytes, were evaluated statistically. - Evaluation criteria:
- Both biological and statistical significances were considered together for evaluation purposes.
A substance is considered positive if there is a significant increase in the number of micronucleated polychromatic erythrocytes for at least one of the time points compared with the concurrent negative control group. A test substance producing no significant increase in the number of micronucleated polychromatic erythrocytes is considered non-mutagenic in this system.
For the assay to be valid, individual and/or group mean values for the positiv control should exeed the laboratory's historical control range. - Statistics:
- A Wilcoxon-Test (one-sided) was evaluated to check the validity of the study. This is done by comparing the number of polychromatic erythrocytes with micronuclei in the positive control with the negative control.
A Wiicoxon-Test (one-sided) was performed for each treatment interval (12h, 24h, 48h) and for polychromatic and normochromatic erythrocytes. These tests are performed sequentially with a multiple level of significance of 5%.
The following comparisons are performed only if there is a difference between the positive control and the negative control (24 h). This was performed with a Wilcoxon-Test (two-sided) with a 5 %-level of significance. Wilcoxon-Tests (two-sided) are performed sequentially for the ratio of polychromatic erythrocytes for each treatment interval (12h, 24h, 48h) at a multiple level of significance of 5 %. The data obtained were also compared with historical controls.
Results and discussion
Test results
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- Doses producing toxicity: 1250 mg/kg, 48 hours after application all animals were free of clinical signs of toxicity.
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: Oral administration of 2000 and 1500 mg per kg body weight resulted in mortality in 1 out of 3 males and 1 out of 3 females (2000 mg/kg bw) and 2 out of 3 males (1500 mg/kg bw), respectively. No mortality occured in the 1000 mg/kg bw dose group.
- Clinical signs of toxicity in test animals:
2000 mg/kg bw dose group:
stilted gait, squatting posture, palpebreal fissure narrow, coat bristling, spontaneous activity decreased, cyanosis
1250 mg/kg bw doese group:
spontaneous activity decreased, palpebreal fissure narrow, palpebreal fissure very narrow, squatting posture, stilted gait and coat bristling
1500 mg/kg bw dose group:
stilted gait, squatting posture, palpebreal fissure narrow, palpebreal fissure closed, coat bristling, spontaneous activity decreased, eyelids adhering and sonoures rales
1000 mg/kg bw dose group:
no clinical signs were observed
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei: The incidence of micronucleated polychromatic and normochromatic erythrocytes in the test group was within the normal range of the negative control groups.
- Ratio of PCE/NCE: The ratio of polychromatic erythrocytes to normocytes remained essentially unaffected by the test compound.
Any other information on results incl. tables
For details on results, please refer to the attached document.
Applicant's summary and conclusion
- Conclusions:
- The test substance did not lead to a substantial increase of micronucleated polychromatic erythrocytes and is not mutagenic in the micronucleus test.
- Executive summary:
The micronucleus test was carried out with the test substance to assess its potential to cause
chromosomal damage (clastogenicity). The test compound was dissolved in DMSO (dried) and was given once as an orally dose of 1250 mg per kg body weight to male and female mice, based on the results of a previous dose range finding assay (see preliminary study). According to the test procedure the animals were killed 12, 24 or 48 hours after administration.
Endoxan® was used as positive control substance and was administered once orally at a dose of 50 mg per kg body weight.
The number of polychromatic and normochromatic erythrocytes containing micronuclei was not increased. The ratio of polychromatic/normochromatic erythrocytes in both male and female animals remained unaffected by the treatment with the test substanceand was statistically not different from the control values.
Endoxan®induced in both males and females a marked statistically significant increase in the number of polychromatic cells with micronuclei, indicating the sensitivity of the test system. The ratio of polychromatic erythrocytes to normocytes was not changed to a significant extent.
Under the conditions of the present study the results indicate that the test substanceis not mutagenic in the micronucleus test.
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.