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EC number: 268-871-1 | CAS number: 68153-22-0
- 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 / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 8th September 2003 to 25th November 2003
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 004
- Report date:
- 2004
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: UK department of Health Committee on Mutagenicity Guidelines for the Mutagenicity Testing of Chemicals.
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Date signed: 13/02/03 Alexander, R.G.
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- Hydrocarbon waxes (petroleum), oxidized
- EC Number:
- 265-205-1
- EC Name:
- Hydrocarbon waxes (petroleum), oxidized
- Cas Number:
- 64743-00-6
- Molecular formula:
- too complex
- IUPAC Name:
- Hydrocarbon waxes (petroleum), oxidized
- Test material form:
- other: greasy solid
- Details on test material:
- - Physical appearance: Brown waxy solid.
- Storage condition of test material: Room temperature in the dark.
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- lymphocytes: Human
- Details on mammalian cell type (if applicable):
- - Average generation time: ~ 17 hours for this donor, determined under typical experimental conditions.
- Medium: Eagle's minimal essential medium with HEPES buffer (MEM), supplemented with L-glutamine, penicillin/streptomycin, amphotericin B and 15% foetal calf serum, at 37ºC with 5% CO2 in air.
- Cells were stimulated to divide by the addition of phytohaemagglutinin (PHA) at 90 µg/ml final concentration. - Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Experiment 1
4 hour exposure with and without S9 mix: 0, 156.25, 312.5, 625, 1250, 1875, 2500 µg/ml
Experiment 2
4 hour exposure with S9 mix: 0, 39.06, 78.13, 156.25, 312.5, 625 and 1250 µg/ml
24 hour exposure without S9 mix: 0, 39.06, 78.13, 156.25, 312.5, 468.75 and 625 µg/ml
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium.
PRELIMINARY STUDY:
A dose range finding study was performed using concentrations of the test material between 19.5 and 5000 µg/ml.
MAIN TEST: The study was carried out in two experiments, utilising four different exposure conditions.
Experiment 1
- The cells were exposed for 4 hours without S9-mix followed by 20 hour culture in media free from the test material, prior to cell harvest.
- The cells were exposed for 4 hours with S9-mix followed by 20 hour culture in media free from the test material, prior to cell harvest.
Experiment 2
- Continuous exposure to the test material for 24 hours prior to cell harvesting.
- The cells were exposed to the test material for 4 hours with S9-mix followed by 20 hour culture in test material free media, prior to cell harvest.
TEST CONDITIONS:
- Preincubation period: 48 hours at 37 ºC, 5% CO2 in humidified air.
- Volume of test material, vehicle or positive control: 0.1 ml
- Total nominal volume: 10 ml.
- Volume and concentration of S9 mix: 1 ml of 20% S9-mix in experiment 1 and 1 ml of 10% S9-mix in experiment 2.
- Expression time (cells in growth medium): 4 hours with the test material and 20 hours without the test material.
- In the case of the continuous exposure the expression time was 24 hours, with the test material.
SPINDLE INHIBITOR (cytogenetic assays): demecolcine, Colcemid 0.1 µg/ml.
STAIN (for cytogenetic assays): 5% Gurrs Giemsa.
NUMBER OF REPLICATIONS: Tests performed in duplicate.
NUMBER OF CELLS EVALUATED: 2000 lymphocytes cell nuclei were counted and the number of cells in methaphase recorded and expressed as the mitotic index and as a percentage of the vehicle control value. - Evaluation criteria:
- Scoring:
Where possible the first 100 consecutive well-spread metaphases from each culture were counted, where there were approximately 50% of cells with aberrations, slide evaluation was terminated at 50 cells. If the cell had 44-48 chromosomes, any gaps, breaks or rearrangements were noted according to the simplified system of Savage (1976).
The frequency of cells with aberrations excluding gaps and the frequency of polyploid cells was compared, where necessary, with the concurrent vehicle control value using Fisher’s Exact test. Samples which showed a statistical increase in aberrations were considered to have given a positive result. - Statistics:
- Fisher’s Exact test.
Results and discussion
Test results
- Key result
- Species / strain:
- lymphocytes: Human
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Experiment 1:
-The test material did not induce a statistically significant increase in the frequency of cells with aberrations either in the absence or presence of metabolic activation.
- The maximum dose level with scoreable metaphases present was 1250µg/ml in the presence and absence of metabolic activation. - The mitotic data confirms the qualitative assessment that the dose-related inhibition of mitotic index was observed and that 33% mitotic inhibition was achieved at 1250 µg/ml in the absence of S9 and 35% in the presence.
- Complete inhibition was observed at 1875 and 2500µg/mlin both exposure groups.
Experiment 2:
-The test material did not induce a statistically significant increase in the frequency of cells with aberrations either in the absence or presence of metabolic activation.
- Qualitative assessment of the slides determined that the maximum dose level with scorable metaphases was 1250µg/ml with S9 mix and 468.75µg/ml without S9 mix in experiment 2.
- The mitotic index data confirms the qualitative observations that a dose-related inhibition of mitotic index was observed and that 67% mitotic inhibition as achieved at 468.75 µg/ml in the absence of S9. In the presence of S9 50% mitotic inhibition was achieved at 1250 µg/ml.
Table 1. Results of Chromosome Aberrations
Experiment No. | With or Without S9 mix | Treatment Group | Dose Level (µg/ml) | Total No. of Aberrations | Frequency of Aberrant Cells (%) | ||
(+ Gaps) | (- Gaps) | (+ Gaps) | (- Gaps) | ||||
1 | Without | Vehicle Control | 2 | 1 | 1 | 1 | |
Test Material | 312.5 | 1 | 1 | 1 | 1 | ||
625 | 3 | 2 | 3 | 2 | |||
1250 | 2 | 1 | 2 | 1 | |||
1875 | TOXIC | ||||||
Positive Control MMC | 0.4 | 39 | 29 | 28 | 22*** | ||
1 | With | Vehicle Control | 7 | 5 | 3 | 1 | |
Test Material | 312.5 | 2 | 2 | 2 | 2 | ||
625 | 1 | 0 | 1 | 0 | |||
1250 | 5 | 3 | 5 | 3 | |||
1875 | TOXIC | ||||||
Positive Control CP | 10 | 89 | 50 | 65 | 39*** | ||
2 | Without | Vehicle Control | 3 | 3 | 3 | 3 | |
Test Material | 156.25 | 1 | 0 | 1 | 0 | ||
312.5 | 1 | 0 | 1 | 0 | |||
468.75 | 1 | 1 | 1 | 1 | |||
625 | TOXIC | ||||||
Positive Control MMC | 0.2 | 72 | 53 | 50 | 39*** | ||
2 | With | Vehicle Control | 2 | 1 | 2 | 1 | |
Test Material | 312.5 | 1 | 0 | 1 | 0 | ||
625 | 2 | 2 | 2 | 2 | |||
1250 | 8 | 3 | 8 | 3 | |||
Positive Control CP | 0.2 | 96 | 73 | 52 | 42*** |
MMC = Mitomycin C
CP = Cyclophosphamide
*** = P < 0.001
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the conditions of the test, the test material was determined to be non-clastogenic to human lymphocytes. There was no statistically significant increase in the frequency of cell aberrations in the absence or presence of metabolic activation. - Executive summary:
In a GLP compliant chromosome aberration study performed according to the standardised guidelines; OECD 473, EU Method B.10 and UK department of Health Committee on Mutagenicity Guidelines for the Mutagenicity Testing of Chemicals, the test material was determined to be non-clastogenic. Under the conditions of the test the human lymphocyte did not produce a statistically significant increase in the frequency of cell aberrations in the absence or presence of metabolic activation. Therefore the test material is considered to be non-mutagenic.
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