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EC number: 210-762-8 | CAS number: 622-97-9
- 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 DNA damage and/or repair study
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 980
- Report date:
- 1980
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 479 (Genetic Toxicology: In Vitro Sister Chromatid Exchange Assay in Mammalian Cells)
- Deviations:
- no
- GLP compliance:
- no
- Type of assay:
- sister chromatid exchange assay in mammalian cells
Test material
- Reference substance name:
- 4-methylstyrene
- EC Number:
- 210-762-8
- EC Name:
- 4-methylstyrene
- Cas Number:
- 622-97-9
- Molecular formula:
- C9H10
- IUPAC Name:
- 1-ethenyl-4-methylbenzene
- Test material form:
- liquid
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- lymphocytes:
- Details on mammalian cell type (if applicable):
- Not specified
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- Trial 1: 1, 10, 25, 50 and 100 nL/mL
Trial 2: 25, 50, 65, 80 and 100 nL/mL (confirmatory assay)
Trial 3: 25, 50, 65, 80 and 100 nL/mL (repeating Trial 2 with new lot of test substance due to suspected improper storage of lot used in both earlier
assays) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO;
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO, 1%
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- 0.1 µgmL
- Details on test system and experimental conditions:
- Whole blood cultures
Human venous blood was drawn into a sterile syringe that contained heparin to prevent clotting. Cultures were initiated by adding 0.6 ml of blood to 9.4 ml pf medium in 25 cm2 plastic flasks standing upright to minimize the surface area. The medium was RPMI 1640 supplemented with 15 % fetal calf serum (FCS), 1% penicillin-streptomycin, and phytohaemagglutinin (PHA-M).
Treatment of cultures with the test substance
Blood was added to culture medium containing PHA and incubated at 37 °C in the dark for 24 h. At this time cells are beginning to grow due to stimulation by PHA. Solutions of the test substance EMS or the solvent control were then added to appreciate cultures, followed by BrdU (final concentration 25 uM) and cultures were re-incubated for a further 46-48 hours (total culture time: 70-72 h). Colcemid was added 2.5 hours before harvest of dividing lymphocytes. Two replicate cultures were used for each treatment in assay.
Lymphocyte fixation
Thee cell suspension was centrifuged, the supernatant discarded, and cells treated with hypotonic KCl (0.075 M) for 3 minutes to swell the cells and get rid of red blood cells. Cells were then washed three times with fixative (methanol/glacial acetic acid, (3:1)) and dropped on to slides to air day. In trial 3 BrdU was added at initiation of cultures, not after the test compound.
Staining for detection of SCE was accomplished by a modified fluorescent plus Giemsa technique (Perry and Wolff 1974 and Goto et al. 1978).
Results and discussion
Test results
- Key result
- Species / strain:
- lymphocytes: Human
- Metabolic activation:
- without
- Genotoxicity:
- ambiguous
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- The solvent control did not show any mutagenic activity. Results for the positive control showed expected mutagenic activity.
Trial 1: A slight, but statistically significant (P<0.001) higher number of sister chromatid exchanges (SCEs) per cell was noted at 50 and 100 nL/mL compared to solvent control, with some evidence of a dose-response.
Trial 2: This assay repeated the first trial, using more concentrations in the range of ‘positives’ from Trial 1. In this assay a slight, but statistically significant (P<0.25 or P<0.001) higher number of SCEs per cell was observed at 50 through 80 nL/mL. However, the difference from solvent control at the highest concentration of 100 nL/mL was not statistically significant and, overall, there was no evidence of a dose-response.
Trial 3: A new lot of the test substance, stored at -20⁰C (instead of room temperature), was used in this trial, using the same concentrations of test substance as in Trial 2 and following essentially the same methodology. A slight, but statistically significant (P<0.001) higher number of SCEs per cell was noted in this assay at concentrations of 65 through 100 nL/mL, without a clear dose-relationship.
Dose-related cell toxicity was noted at all test substance concentrations, as evidenced by a reduction in the frequency of dividing cells (mitotic index) and cell cycle delay.
Any other information on results incl. tables
Treatment |
Mitotic index (% 500 cells) |
% of cells at first second and third metaphase |
||
Solvent control |
1.2 % |
26 |
58 |
16 |
25 nl/ml |
0.4 % |
42 |
51 |
7 |
100 nl/ml |
0.6 % |
66 |
34 |
0 |
Applicant's summary and conclusion
- Conclusions:
- Under the study conditions, dose-related cell toxicity was noted at all test substance concentrations, as evidenced by a reduction in the frequency of dividing cells (mitotic index) and cell cycle delay. A slight but statistically significant and reproducible higher number of SCEs was observed in cultured human lymphocytes treated with test substance over a concentration range of 50 to 100 nL/mL. The maximum response in cultures treated with test material was approximately 84% over the solvent control. Given that these data did not meet the evaluation criteria for a ‘positive’ result, with a less than a 2-fold increase in SCEs over solvent control and without a clear dose-response, the test substance was designated as ‘weakly positive’.
- Executive summary:
A study using cultured human lymphocytes was conducted to determine the potential of the test substance to induce sister chromatid exchanges (SCEs) in vitro, according to a method similar to OECD Guideline 479. DMSO (1%) was used as the solvent control and the positive control was ethylmethanesulfonate. Three trials were performed, using an overall concentration range of 1 to 100 nL/mL. The solvent control did not show any activity, results for the positive control showed expected increases in SCEs. Under the study conditions, dose-related cell toxicity was noted at all test substance concentrations, as evidenced by a reduction in the frequency of dividing cells (mitotic index) and cell cycle delay. A slight but statistically significant and reproducible higher number of SCEs was observed in cultured human lymphocytes treated with test substance over a concentration range of 50 to 100 nL/mL. The maximum response in cultures treated with test material was approximately 84% over the solvent control. Given that these data did not meet the evaluation criteria for a ‘positive’ result, with a less than a 2-fold increase in SCEs over solvent control and without a clear dose-response, the test substance was designated as ‘weakly positive’ (Galloway, 1980).
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