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EC number: 606-712-3 | CAS number: 211450-54-3
- 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 cytogenicity / micronucleus study
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 013
- Report date:
- 2013
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline for Testing of Chemicals, July 22, 2010, Guideline No. 487 “In vitro Mammalian Cell Micronucleus Test".
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Harlan Cytotest Cell Research GmbH (Harlan CCR), In den Leppsteinswiesen 19, 64380 Rossdorf, Germany
- Type of assay:
- in vitro mammalian cell micronucleus test
Test material
- Reference substance name:
- Soybean oil, epoxidized, reaction products with methanol
- EC Number:
- 287-837-7
- EC Name:
- Soybean oil, epoxidized, reaction products with methanol
- Cas Number:
- 85586-35-2
- Molecular formula:
- C H4 O . Unspecified
- IUPAC Name:
- Epoxidized Soybean Oil, reaction products with methanol
- Details on test material:
- - Test Item: Soybean oil, epoxidized, reaction products with methanol
- BASF Test Item No.: 12/0114-1
- Batch Number: CE61910022
- Purity: 100%
- Expiration Date: December 2013
- Physical state, appearance: Liquid, yellow
- Storage conditions: At room temperature
- tability in Solvent: Not indicated by the Sponsor
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- Stocks of the V79 cells (obtained from LMP; Technical University Darmstadt, 64287 Darmstadt, Germany) are stored in liquid nitrogen in the cell bank. Before freezing each batch is screened for mycoplasm contamination and checked for karyotype stability.
- Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/β-naphthoflavone induced rat liver S9 mix
- Test concentrations with justification for top dose:
- See "any other information on materials and methods"
- Vehicle / solvent:
- The final concentration of DMSO in the culture medium was 0.5 % (v/v). The solvent was chosen due to its solubility properties and its relative non-toxicity to the cell cultures.
Controlsopen allclose all
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- With metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- Without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Griseofulvin
- Remarks:
- Without metabolic activation
- Details on test system and experimental conditions:
- DOSE-SELECTION:
With respect to the solubility of the test item, 4000.0 µg/mL of Soybean oil, epoxidized, reaction products with methanol was applied as top concentration for treatment of the cultures in the pre-test. Test item concentrations between 7.8 and 4000.0 µg/mL (with and without S9 mix) were chosen for the evaluation of cytotoxicity. No precipitation of the test item was observed. Since the cultures fulfilled the requirements for cytogenetic evaluation in the presence of S9 mix, this preliminary test was designated Experiment IA. The experimental part without S9 mix was repeated with the same top concentration due to the low response of the positive control MMC and designated Experiment IB. Since no clear cytotoxicity and test item precipitation was observed in Experiment IA and IB, the same concentrations were applied in Experiment II.
EXPERIMENTAL PERFORMANCE:
- Culture Medium and Conditions: For seeding and treatment of the cell cultures the culture medium was MEM (minimal essential medium) containing Hank’s salts, glutamine, Hepes (25 mM) and 10 % (v/v) fetal bovine serum (FBS). Additionally, the medium was supplemented with penicillin/streptomycin (100 U/mL/100 µg/mL). All cultures were incubated at 37°C in a humidified atmosphere with 1.5 % CO2 (98.5 % air).
- Seeding of the Cultures: Exponentially growing stock cultures more than 50 % confluent were rinsed with Ca-Mg-free salt solution containing 8000 mg/L NaCl, 200 mg/L KCl, 200 mg/L KH2PO4 and 150 mg/L Na2HPO4. Afterwards the cells were treated with trypsin-EDTA-solution at 37°C for approx. 5 minutes. Then, by adding complete culture medium including 10 % (v/v) FBS the enzymatic treatment was stopped and a single cell suspension was prepared. The trypsin concentration for all subculturing steps was 0.25 % (w/v) in Ca-Mg-free salt solution. The cells were seeded into Quadriperm dishes, which contained microscopic slides. The cells were seeded into Quadriperm dishes, which contained microscopic slides. Into each chamber 1.0 x 10^5 – 1.5 x 10^5 cells were seeded with regard to the preparation time.
TREATMENT:
- Exposure period 4 hours: The culture medium of exponentially growing cell cultures was replaced with serum-free medium containing the test item. For the treatment with metabolic activation 50 µL S9 mix per mL medium was added. Concurrent solvent and positive controls were performed. After 4 hours the cultures were washed twice with "Saline G" (pH 7.2) containing 8000 mg/L NaCl, 400 mg/L KCl, 1100 mg/L glucose • H2O, 192 mg/L Na2HPO4 • 2 H2O and 150 mg/L KH2PO4. Then the cells were cultured in complete medium containing 10 % (v/v) FBS for the remaining culture time of 20 hours.
- Exposure period 24 hours: The culture medium of exponentially growing cell cultures was replaced with complete medium containing 10 % (v/v) FBS including the test item without S9 mix. The medium was not changed until preparation of the cells. Concurrent solvent and positive controls were performed.
PREPARATION OF THE CULTURES:
For the micronucleus analysis, 24 hours after the start of the exposure, the cells were treated on the slides in the chambers of the quadriperm dishes with deionised water for 1 to 1.5 min at 37°C. Afterwards the cells were fixed twice with a solution containing 3 parts ethanol, 1 part acetic acid and 1.25 % (v/v) formaldehyde. After preparation the cells were stained with Giemsa and labelled with a computer-generated random code to prevent scorer bias.
ANALYSIS OF MICRONUCLEI AND CYTOTOXICITY:
Evaluation of the cultures was performed manually using NIKON microscopes with 40x objectives. The micronuclei were counted in cells showing a clearly visible cytoplasm area. The criteria for the evaluation of micronuclei are described in the publication of Countryman and Heddle (1976). Briefly the micronuclei were stained in the same way as the main nucleus. The area of the micronucleus did not extend the third part of the area of the main nucleus. 1000 cells in two parallel cultures were scored for micronuclei, so that at least 2000 cells from clones with 2-8 cells were analysed per test group. The frequency of micronucleated cells was reported as % micronucleated cells. Cytotoxicity was assessed via counting the number of clones consisting of 1 cell (c1), 2 cells (c2), 3-4 cells (c4), and 5-8 cells (c8) among the cells that were scored for the presence of micronuclei. These clusters represented the cells that have divided 1, 2, or 3 times within the experiment. From these data, a proliferation index (PI) was calculated. Only those cultures were evaluated which showed a PI > 1.3, in order to guarantee for a sufficient cell proliferation during treatment and recovery. - Evaluation criteria:
- ACCEPTABILITY OF THE ASSAY:
The micronucleus assay is considered acceptable if it meets the following criteria:
a) The number of micronucleated cells carrying one or more micronuclei found in the solvent controls falls within the range of the historical laboratory control data range.
b) The positive control substances produce a significant increase (at least two-fold the respective control value) in the number of micronucleated cells exceeding the historical laboratory solvent control data range.
EVALUATION OF RESULTS:
A test item can be classified as mutagenic if:
- the number of micronucleated cells is not in the range of the historical control data and
- either a statistically significant concentration-related increase in three test groups or a significant increase of micronucleated cells in at least one test group is observed.
A test item can be classified as non-mutagenic if:
- the number of micronucleated cells in all evaluated test groups is in the range of the historical control data and
- no statistically significant concentration-related increase in the number of micronucleated cells is observed. - Statistics:
- Statistical significance was confirmed by means of the Chi square test
Results and discussion
Test results
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- No precipitation of the test item in the culture medium was observed. Phase separation was observed in Experiment IA, IB and II in the presence of S9 mix at 125.0 µL/mL and above. In Experiment II in the absence of S9 mix phase separation was observed at 250.0 µg/mL and above. No relevant influence on pH value was observed. The osmolarity was slightly increased at the highest applied concentration in Experiment II. In the absence and presence of metabolic activation no clear cytotoxicity measured as reduced proliferation index was observed. No mutagenicity was observed at the concentrations evaluated. In Experiment IA and IB in the absence and presence of S9 mix two statistically significant increases were observed after treatment with 1000 µg/mL (1.20 and 1.30 %). The values are in the range of the laboratory historical control data (0.15-1.50 % and 0.05-1.70 % micronucleated cells, respectively) and therefore biologically irrelevant. In Experiment II in the presence of S9 mix two statistically significant increases were observed after treatment with 62.5 and 2000.0 µg/mL (1.55 and 1.45 %). The values are in the range of the laboratory historical control data (0.05 - 1.70 % micronucleated cells) and therefore biologically irrelevant. In the presence of S9 mix one single value (1.60 %) exceeded the range of the laboratory historical control data (0.05-1.50 % micronucleated cells) after treatment with 1000.0 µg/mL. The value is not statistically significant and therefore biologically irrelevant. Mitomycin C (0.1 µg/mL), Griseofulvin (8.0 µg/mL) or CPA (15.0 µg/mL) were used as positive controls and showed a distinct increase in the percentage of micronucleated cells.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Applicant's summary and conclusion
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