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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 266-411-4 | CAS number: 66576-71-4
- 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
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 03 November to 19 December 2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Reference substance name:
- Isopropyl 2-methylbutyrate
- EC Number:
- 266-411-4
- EC Name:
- Isopropyl 2-methylbutyrate
- Cas Number:
- 66576-71-4
- Molecular formula:
- C8H16O2
- IUPAC Name:
- isopropyl 2-methylbutyrate
- Test material form:
- liquid
- Details on test material:
- - Name of test material (as cited in study report): Isopropyl 2-methylbutyrate
- Substance type: Organic
- Physical state: Liquid
- Analytical purity: 99.9%
- Purity test date:
- Lot/batch No.: H4-K-27
- Expiration date of the lot/batch: 17 August 2018
- Stability under test conditions: Detailed data regarding stability during storage and under test conditions are not available. It is however expected that no gross degradation occurs under the specified storage conditions as specified by the sponsor, or over the duration of the study when dissolved in DMSO, the selected solvent for the study (duration of treatment ca. 4 hours).
- Storage condition of test material: Ambient condition, protected from light
Constituent 1
Method
- Target gene:
- hprt
Species / strain
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Growth medium, media containing selective agent, media without selective agent and HAT medium
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9
- Test concentrations with justification for top dose:
- Main test concentration selection (in the absence and presence of S9): 45.125, 90.25, 180.5, 361, 722 and 1444 µg/Ml (10 Mm). Concentrations based on the cytotoxicity test performed in the absence and presence of metabolic activation at the same concentrations as above
- Vehicle / solvent:
- Vehicle used: DMSO
- Justification for choice of solvent/vehicle: The test item was insoluble in sterile distilled water, however it was soluble in dimethyl sulfoxide at concentrations >1444 µg/mL (10Mm), the guideline limit concentration. Volume used for treatment (250 µL per 25 mL medium)
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension;
DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 8 days
- Selection time (if incubation with a selection agent): 8 days
- Fixation time (start of exposure up to fixation or harvest of cells): 16 days
SELECTION AGENT (mutation assays): 2-amino-6-mercaptopurine (6-thioguanine) at 5 µg/mL -alpha-MEM without nucleosides
NUMBER OF REPLICATIONS: Two replicate flasks per concentration
DETERMINATION OF CYTOTOXICITY
- Method: Relative cloning efficiency. - Evaluation criteria:
- Assay Acceptance Criteria
A mutation assay was considered acceptable if:
A minimum 60% absolute cloning efficiency in negative controls (DMSO) and a spontaneous mutant frequency less than 20 per 106 clonable cells.
Positive controls induce a significant increase in the mutant frequency above the concurrent negative control.
Assay Evaluation Criteria
A test item was considered positive if:
The test item caused a concentration-related biologically significant increase in mutant frequency in comparison with concurrent negative control and the test item causes a three-fold increase in the number of 6-thioguanine resistant colonies relative to concurrent negative control and such increases were statistically significant and outside the laboratory historical negative (DMSO) control range.
A net increase in mutant colonies of treated above the concurrent control was observed in at least two of the concentrations tested.
Clear negative results were not confirmed by a repeat test (short duration), as per revised OECD guideline. - Statistics:
- Weighted regression analysis was performed to evaluate the dose response relationship on isopropyl 2-methylbutyrate treatment groups against the negative control group. Statistical analysis was not performed for the positive controls.
Results and discussion
Test results
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: strain/cell type: Chinese Hamster Ovary (CHO)-K1 Cell Line’
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
No precipitation was observed at 0 and 4 hours prior to the cytotoxicity test. No biologically relevant influence of Isopropyl 2-methylbutyrate on pH or osmolality were observed both in the absence and presence of metabolic activation during the main study.
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results:
negative with metabolic activation
negative without metabolic activation
Isopropyl 2-methylbutyrate does not have potential to induce gene mutations at the hprt locus of CHO-K1 cells, both in the absence and presence of metabolic activation under the experimental conditions described.
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.