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EC number: 252-667-4 | CAS number: 35674-65-8
- 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 vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: According to international guideline(s), GLP-compliant, performed in recognized contract research organization, no restrictions, fully adequate for assessment.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 003
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Version / remarks:
- of 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: U.S. Food and Drug Administration (FDA), Redbook 2000: Toxicological Principles for the Safety of Food Ingredients, IV.C.1.c Mouse Lymphoma Thymidine Kinase Gene Mutation Assay, 2001
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Test material form:
- solid: particulate/powder
- Remarks:
- migrated information: powder
Constituent 1
Method
- Target gene:
- TK
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- Cell line used: L5178Y tk +/- clone 3.7.2C
The chromosome number of these cells is 40 (stable aneuploid karyotype, 2n = 40). The cells were obtained from MRC Cell Mutation Unit, University of Sussex, Falmer, Brighton, East Sussex, BN1 9RR, UK.
- Type and identity of media: RPMI 1640 medium (with HEPES and Glutamax-I) supplemented with heat-inactivated horse serum (10% v/v for growing in flasks, and 20% for growing in microtiter plates), sodium pyruvate and penicillin/streptomycin.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: a karyotype check was carried out in 1995
- Periodically "cleansed" against high spontaneous background: no data
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix (liver fraction of Aroclor 1254-induced male Wistar rats) supplemented with a NADPH-generating system)
- Test concentrations with justification for top dose:
- ASSESSMENT OF CYTOGENICITY AND MUTAGENICITY
The highest concentrations tested were 108 and 500 µg/mL in the absence and presence of S9-mix, respectively.
The highest concentrations evaluated for mutagenicity were 8.4 and 75 µg/mL in the absence and presence of S9-mix, respectively.
- Without metabolic activation:
First assay: 108, 65, 39, 23, 14, 8.4*, 5.0*, 3.0*, 1.8*, 1.1*, 0.65*, 0.39*, 0.24*, and 0.14⁺ µg/mL, single cultures
Second assay: 15, 12, 9.6, 8.2⁺, 6.9*, 5.9*, 5.0*, 4.0*, 3.2, 2.4*, 1.8, 1.4*, 0.68*, 0.34, 0.17*, and 0.08* µg/mL, single cultures
- With metabolic activation:
First assay: 500, 300, 180, 108, 65⁺, 39*, 23*, 14*, 8.4*, 5.0*, 3.0*, 1.8*, 1.1*, and 0.65 µg/mL, single cultures
Second assay: 30⁺, 24*, 19*, 16*, 14, 12*, 10, 8.0*, 6.4, 4.8*, 3.6, 2.7⁺, 1.4⁺, 0.68, and 0.34 µg/mL, single cultures
Third assay: 75*, 60*, 48*, 38*, 31, 25*, 20, and 16 µg/mL, duplicate cultures
* Assessed for mutagenicity (both replicates of the third assay)
⁺ Scored for small and large colonies
Dose related slight turbidity was observed at the start of treatment at concentrations above 23 µg/mL; at the end of treatment, slight turbidity was observed at concentrations above 8.4 µg/mL. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: tetrahydrofuran (THF)
- Justification for choice of solvent/vehicle: The test material could be suspended in THF up to 50 mg/mL, resulting in a homogeneous white turbid suspension
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 3-methylcholanthrene
- methylmethanesulfonate
- Remarks:
- MMS and MCA were used as positive control substances in the absence and presence of the S9-mix, respectively. Final concnetartions: 0.1 mM (11 µg/mL, MMS) and 10 µg/mL (MCA).
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours (with metabolic activation) , 24 hours (without metabolic activation)
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 10-14 days
SELECTION AGENT (mutation assays): trifluorothymidine (TFT), final concentration 4 µg/mL
NUMBER OF REPLICATIONS:
- Exposure phase:
first and second assay and positive controls: single cultures;
third assay and vehicle controls: duplicates cultures.
- Assessment of cloning efficiency and mutant potential:
two 96-well plates from each culture (four wells if a reduced cloning efficiency (<50%) was expected)
DETERMINATION OF CYTOTOXICITY
- Method: initial cell yield, relative total growth (RSG) and relative total growth (RTG)
OTHER EXAMINATIONS:
- Scoring of small and large colonies (negative and positive controls and of some test material dose levels)
GROWTH CONDITIONS
- at ca. 37 °C and ca. 5% CO2 in a humidified incubator - Evaluation criteria:
- A response was considered to be positive if the induced mutant frequency (mutant frequency of the test substance minus that of the vehicle negative control) was more than 100 mutants per 1,000,000 clonable cells*. A response was considered to be equivocal if the induced mutant frequency was more than 50 mutants per 1,000,000 clonable cells. Any apparent increase in mutant frequency at concentrations of the test substance causing more than 90% cytotoxicity was considered to be an artefact and not indicative of genotoxicity.
* Aaron et al, 1994, Mutation Res. 312:235-239; Clive et al., 1995, Environ. Molec. Mutagen. 25:165-168
The test substance was considered to be mutagenic in the gene mutation test at the TK -locus if a concentration-related increase in mutant frequency was observed, or if a reproducible positive response for at least one of the test substance concentrations was observed.
The test substance was considered not to be mutagenic in the gene mutation test at the TK-locus if it produced neither a dose-related increase in the mutant frequency nor a reproducible positive response at any of the test points.
Both numerical significance and biological relevance were considered together in the evaluation. - Statistics:
- No statistical analysis was performed.
Results and discussion
Test results
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- see 'Additional information on results'
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- ADDITIONAL INFORMATION ON CYTOTOXICITY:
The test material was cytotoxic to the L5178Y cells in both the absence and the presence of S9-mix. The initial cell yield, relative suspension growth (RSG), and relative total growth (RTG) were decreased above concentrations of 3 µg/mL and 30 µg/mL in the absence and presence of S9-mix, respectively. The RTG values (according to OECD and FDA guidelines) at the highest concentrations evaluated for mutagenicity were below 20%.
Any other information on results incl. tables
In the first and second assay, a slight increase of 53 -75 mutants per 10E6 clonable cells compared to the negative control was observed at three single concentrations in both the absence and presence of metabolic activation. In the third assay no increase of the mutant frequency was observed at any dose level, either at toxic or non-toxic dose levels. Since the observed increases of the mutant frequencies were not reproducible they were considered as accidental occurrence and not indicative for mutagenicity.
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
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.
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