Registration Dossier
Registration Dossier
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: 209-400-1 | CAS number: 576-26-1
- 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:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Well documented study following OECD 474 and GLP. The reliability of this study for the substance tested is a K1, but in application of read-across to a different substance ECHA’s guidance specifies that the score can be a maximum of K2. A justrification for the read-across is provided.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Report date:
- 2002
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
Test material
- Reference substance name:
- structural analogue
- IUPAC Name:
- structural analogue
- Reference substance name:
- 2,4,6-trimethylphenol
- EC Number:
- 208-419-2
- EC Name:
- 2,4,6-trimethylphenol
- Cas Number:
- 527-60-6
- Molecular formula:
- C9H12O
- IUPAC Name:
- 2,4,6-trimethylphenol
- Details on test material:
- - Storage condition of test material: stored at room temperature protected from light and moisture
Constituent 1
Constituent 2
Test animals
- Species:
- mouse
- Strain:
- ICR
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: HArlan- Sprague-Dawley, Inc., Frederick, Md
- Age at study initiation: Mice were 6-8 weeks old for each phase of the study
- Weight at study initiation: Animal body weights were within the following ranges at randomization for each of these phases:
- Pilot phase: Males: 26.9-30 g, Females: 27.2-29.0 g
- Toxicity phase: Males: 24.7 - 32.2 g, Females: 25.1-29.0 g
- Definitive Micronucleus assay: Males: 26.9-32.1 g, Females: 24.7-29.0 g
- Assigned to test groups randomly: yes
- Fasting period before study:
- Housing: 5/cage in polycarbonate cages. Heat treated hardwood chips were used for bedding.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: no less than 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72 degrees F +/- 3 degrees F
- Humidity (%): 50% +/- 20%
- Photoperiod (hrs dark / hrs light):12 hour light/dark cycle
Administration / exposure
- Route of administration:
- intraperitoneal
- Vehicle:
- - Vehicle(s)/solvent(s) used: Corn Oil form Sigma-Aldrich
- Details on exposure:
- The test substance-vehicle mixture, the vehicle alone, or the positive control substance were administered as a single intraperitoneal injection at a dose volume of 20 mL/kg body weight. Mice were observed after dose administration for signs of toxicity.
- Duration of treatment / exposure:
- The test substance was dosed once by an intraperitoneal injection.
- Frequency of treatment:
- The test substance was dosed once by an intraperitoneal injection.
- Post exposure period:
- 24 and/or 48 hours
Doses / concentrations
- Remarks:
- Doses / Concentrations:
125, 250, or 500 mg/kg
Basis:
nominal conc.
- No. of animals per sex per dose:
- 5/sex/dose for the low dose, mid dose, and positive control. 10/sex/dose for the high dose and vehicle control. 5 additional animals/sex were dosed with the highest dose (500 mg/kg) to ensure the avaiability of 5 animals per sex per dose group
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Cyclophosphamide monohydrate dissolved in sterile water at a concentration of 2.5 mg/mL for a dose of 50 mg/kg.
Examinations
- Tissues and cell types examined:
- Bone marrow cells, polychromatic erythrocytes (PCEs) and normochromatic erythrocytes (NCEs) were analyzed for the presence of micronuclei.
- Details of tissue and slide preparation:
- Marrow Collection and Slide Preparation
Scoring for Micronuclei
CRITERIA FOR DOSE SELECTION:
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
5-10 mice /sex dosed with either corn oil, 125, 250, or 500 mg/kg of the test substance in corn oil , or 50 mg/kg cyclophosphamide (positive control) were sacrificed after 24 hours and sacrificed for the evaluated of micronuclei in the bone marrow. An additional 5 mice/sex dosed with either corn oil or 500 mg/kg of the test substance were also sacrified after 48 hours for the evaluation of micronuclei in the bone marrow .
DETAILS OF SLIDE PREPARATION:
At the scheduled sacrifice times, five mice per sex per treatment were sacrificed by CO2 asphyxiation. Immediately following sacrifice, the femurs were distally exposed, cut just above the knee, and the bone marrow was aspirated into a syringe containing fetal bovine serum. The bone marrow cells were transferred to a capped centrifuge tube containing approximately 1 mL fetal bovine serum. The bone marrow cells were pelleted by centrifugation at approximately 100 x g for five minutes and the supernatant was drawn off, leaving a small amount of serum with the remaining cell pellet. The cells were resuspended and a small drop of bone marrow suspension was spread onto a clean glass slide. Two slides were prepared from each mouse. The slides were fixed in methanol, stained with May-Gruenwald-Giemsa and permanently mounted.
METHOD OF ANALYSIS:
Micronuclei are round, darkly-staining nuclear fragments with a sharp contour and diameters usually from 1/20 to 1/5 of an erythrocyte. Micronuclei can occur in both PCEs (MPCEs) and NCEs (MNCEs).
2000 polychromatic erythrocytes per animal were scored for the presence of micronuclei. The number of MNCEs in the field of 2000 PCEs was enumerated for each animal in order to assess the quality of the differential staining procedure. The proportion of PCEs to total erythrocytes was also recorded per 1000 erythrocytes. - Evaluation criteria:
- The incidence of MPCEs per 2000 PCEs was determined for each mouse and treatment group. Statistical significance was determined using the Kastenbaum-Bowman tables, which are based on the binomial distribution (Kastenbaum and Bowman, 1970). All analyses were performed separately for each sex and sampling time.
In order to quantify the proliferation state of the bone marrow as indicator of bone marrow toxicity, the proportion of PCEs to total erythrocytes was determined for each animal and treatment group. - Statistics:
- All conclusions were based on sound scientific judgement; however, as a guide to interpretation of the data, the test article was considered to induce a positive response if a dose-responsive increase in MPCEs was observed and one or more doses were statistically elevated relative to the vehicle control (p<=0.05, Kastenbaum-Bowman Tables) at any sampling time. However, values that were statistically significant but did not exceed the range of historical negative or vehicle controls were judged as not biologically significant. If a single treatment group was significantly elevated at one sacrifice time with no evidence of a dose-response or if there was evidence of a dose response with no evidence of a significant increase in any treated group relative to the control group, the Sponsor MPCEs above the concurrent vehicle control values and no evidence of dose response were observed at any sampling time.
Results and discussion
Test results
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- at highest dose tested
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
In the pilot study, all male and female mice at 2000 mg/kg were found dead on the day of dosing. Clinical signs, consisting of piloerection, were observed in males at 1, 10, 100 and 1000 mg/kg. In addition, loss in body weight of approximately 10% was observed in male mice at 1000 mg/kg. Due to mortality at 2000 mg/kg, a toxicity study was performed.
In the toxicity study, mortality was observed in 2/5 males and 2/5 females at 1000 mg/kg and in all males and females at doses >=1000 mg/kg of the test substance. Clinical signs following dose administration included: piloerection, prostration and irregular breathing in males and females at 500 and 1000 mg/kg and lethargy in males at 1000 mg/kg of the test substance. Loss in body weight from 2-14% was observed in male and female mice at 500 and 1000 mg/kg. Based upon these results, the high dose for the micronucleus test was set at 500 mg/kg, which was estimated to be the maximum tolerated dose.
RESULTS OF DEFINITIVE STUDY
No mortality occurred at any dose level during the course of the study. Clinical signs following dose administration included: piloerection in male and female mice at all doses and lethargy in males and females at 250 mg/kg. In addition, prostration and irregular breathing were observed in males and females at 500 mg/kg.
Bone marrow cells (polychromatic erythrocytes, PCEs and normochromatic erythrocytes, NCEs), collected 24 and 48 hours after treatment were examined microscopically for presence of micronuclei (MPCEs or MNCEs). No appreciable reductions in the ratio of PCEs to total erythrocytes was observed in the test article-treated groups relative to the vehicle control groups suggesting that the test article did not inhibit erythropoiesis. The number of MPCEs per 10000 PCEs in test article-treated groups was not statistically increased relative to the respective vehicle controls in either male or female mice, regardless of dose level or bone marrow collection time (p>0.05, Kastenbaum-Bowman Tables). In addition, no appreciable increase in the number of MNCEs in the field of 2000 PCEs per animal was found indicating that an optimal differential staining was achieved.
Any other information on results incl. tables
Cyclophosphamide induced a significant increase in MPCEs in both male and female mice (p<=0.05, Kastenbaum-Bowman Tables). The negative and positive controls were consistent with the historical control data, indicating that there was no problem with the test system or the quality of the test.
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information): negative
Under the conditions of this test, the test substance did not exhibit any mutagenic activity in this in-vivo micronucleus assay. A justification for the read-across has been provided.
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.
