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EC number: 812-653-6 | CAS number: 68441-94-1
- 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
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions (shorter exposure period. Lack of data on test substance, no positive controls for 40 h time point)
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 995
- Report date:
- 1995
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Version / remarks:
- adopted in 1997
- Deviations:
- yes
- Remarks:
- (in both experiments, cultures without metabolic activation were exposed to the test substance for about 16 h, no positive control for the 40 h time point)
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- 189200-42-8
- Cas Number:
- 189200-42-8
- IUPAC Name:
- 189200-42-8
Constituent 1
Method
- Target gene:
- Not applicable.
Species / strain
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: McCoy's 5A Medium containing 10% (v/v) fetal bovine serum and 2 mM L-glutamine
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes - Additional strain / cell type characteristics:
- other: WBL clone
- Metabolic activation:
- with and without
- Metabolic activation system:
- Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of Sprague-Dawley rats treated with Aroclor 1254.
- Test concentrations with justification for top dose:
- 40, 80 and 160 µg/mL with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: Based on results of a solubility test, acetone was selected as the vehicle. The test substance was not soluble in water or dimethyl sulfoxide at any of the concentrations (10, 25, 50% (v/v)) tested. The test substance was soluble as a 50% mixture in acetone.
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: - S9: N-Methyl-N-Nitro-N-Nitrosoguanidine (MNNG), 0.6 µg/mL (v/v) in acetone; + S9: 7,12-Dimethylbenz[a]anthracene (DMBA), 10 µg/mL (v/v) in acetone
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration:
+S9: ca. 3 h (± 0.5 h)
- S9: ca. 16 h (± 0.5 h)
- Fixation time (start of exposure up to fixation or harvest of cells): ca. 16 h (± 0.5 h); second experiment - ca. 16 and 40 h (± 0.5 h)
SPINDLE INHIBITOR (cytogenetic assays): 0.2 mL Colcemid® (10 mg/mL (v/v) in cell culture medium)
STAIN (for cytogenetic assays): 5% Giemsa
NUMBER OF REPLICATIONS: 2 replications (16 h) and 1 replication (40 h), respectively
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 1000 cells - Evaluation criteria:
- A test substance was considered positive in the chromosome aberration test if:
1. A statistically significant dose-related increase in the percentage of aberrant cells and in at least one of the treatment groups, the percentage of aberrant cells exceeds 5%. OR
2. A reproducible and statistically significant response for at least one of the treatment groups is observed. In addition, the mean percentage of aberrant cells exceeds 5%.
A positive result indicates that under the test conditions the test substance induces chromosomal aberrations in cultured mammalian somatic cells.
If neither of the above conditions exist, the test substance is considered nonmutagenic or negative for inducing chromosomal aberrations in this system.
Results and discussion
Test results
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: The test substance is not soluble in water, therefore it was dissolved in acetone.
- Precipitation: Final concentrations of the test substance in medium of 10, 20, 39, 78, 156, 313, 625, 1250 and 2500 µg/mL were tested by visual and microscopic methods for precipitation immediately, 30 minutes and 3 h after dosing. Traces of the test substance were observed microscopically at all test concentrations equal to or greater than 78 µg/mL. Therefore, the upper limit of the culture medium solubility of the test substance was considered to be between 39 and 78 µg/mL. Based on these results, the study director selected the following concentrations for the toxicity pretest: 0.625, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 µg/mL.
In the main experiments, slight precipitation was observed in the second experiment after 16 h at 160 µg/mL without metabolic activation. Precipitation was not noted at any other harvest of a 160 µg/mL culture.
RANGE-FINDING/SCREENING STUDIES: To determine a concentration selection for the aberration assay, a toxicity pretest was conducted with concentrations of 0.625, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 µg/mL of the test substance with and without metabolic activation. The concentrations tested were based on the results of a culture medium solubility test. The cultures with metabolic activation were treated for 3 h (± 0.5 h). The cultures without metabolic activation were treated until 2-3 h prior to harvest. All cultures were harvested about 16 h from the beginning of treatment. After harvest, the number of cells that survived treatment were counted using a hemacytometer to evaluate cytotoxicity and the mitotic indices (number of mitotic cells per 1000 total cells) were determined to evaluate cell cycle suppression. The selected concentrations for the aberration assay were based on the results of the cell count data and mitotic index data. The highest reduction in cell survival was observed at 160 µg/mL without metabolic acvtivation, where reduction in viability of 37% was noted. Other less notable reductions in cell survival were noted (see table 3), but were not indicative of a concentration-related trend. Based on these results, the concentrations selected for the aberration assay were 40, 80 and 160 µg/mL. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
Table 1. Test results of experiment 1
Test item |
Concentration |
Mitotic Index |
Aberrant cells |
Aberration frequency |
|
in µg/mL |
in % |
in % |
in % |
Exposure period 16 h, fixation time 16 h, without S9 mix |
||||
vehicle |
0.5% (v/v) |
6.8 |
0.5 |
0.5 |
MNNG |
0.6 |
6.2 |
22.5** |
24.5 |
Test substance |
40 |
5.6 |
0.5 |
0.5 |
80 |
6.5 |
0.5 |
0.5 |
|
160 |
7.2 |
1.0 |
1.0 |
|
Exposure period 3 h, fixation time 16 h, with S9 mix |
||||
Acetone |
0.5% (v/v) |
5.5 |
1.0 |
1.0 |
DMBA |
10 |
2.6 |
33.5** |
42.0 |
Test substance |
40 |
5.3 |
1.5 |
1.5 |
80 |
6.3 |
0.5 |
0.5 |
|
160 |
4.6 |
2.0 |
2.0 |
**statistically significantly higher than vehicle control (p<0.001)
MNNG: N-Methyl-N-Nitro-N-Nitrosoguanidine; DMBA: 7,12-Dimethylbenz[a]anthracene (positive controls)
Table 2. Test results of experiment 2
Test item |
Concentration |
Mitotic Index |
Aberrant cells |
Aberration frequency |
|
in µg/mL |
in % |
in % |
in % |
Exposure period 16 h, fixation time 16 h, without S9 mix |
||||
vehicle |
0.5% (v/v) |
7.2 |
1.0 |
1.0 |
MNNG |
0.6 |
5.4 |
17.5** |
17.5 |
Test substance |
40 |
7.1 |
1.0 |
1.0 |
80 |
5.4 |
0.5 |
0.5 |
|
160 |
7.1 |
2.5 |
2.5 |
|
Exposure period 3 h, fixation time 16 h, with S9 mix |
||||
Acetone |
0.5% (v/v) |
2.2 |
0.0 |
0.0 |
DMBA |
10 |
4.5 |
33.0** |
43.0 |
Test substance |
40 |
2.2 |
1.0 |
1.0 |
80 |
2.4 |
2.0 |
2.0 |
|
160 |
2.0 |
1.5 |
1.5 |
|
Exposure period 16 h, fixation time 40 h, without S9 mix |
||||
Acetone |
0.5% (v/v) |
3.4 |
2.5 |
2.0 |
MNNG # |
0.6 |
--- |
--- |
--- |
Test substance |
40 |
3.0 |
4.0 |
4.5 |
80 |
2.2 |
3.0 |
3.0 |
|
160 |
3.4 |
2.0 |
2.0 |
|
Exposure period 3 h, fixation time 40 h, with S9 mix |
||||
Acetone |
0.5% (v/v) |
4.8 |
2.5 |
2.5 |
DMBA # |
10 |
--- |
--- |
--- |
Test substance |
40 |
5.4 |
2.0 |
2.0 |
80 |
4.8 |
2.0 |
2.0 |
|
160 |
5.0 |
0.5 |
0.5 |
**statistically significantly higher than vehicle control (p<0.001)
MNNG: N-Methyl-N-Nitro-N-Nitrosoguanidine; DMBA: 7,12-Dimethylbenz[a]anthracene (positive controls)
# According to the study report, positive controls were not required for the 40 h harvest.
Table 3. Toxicity pretest results
Treatment Group |
Cell Survival in %* |
|
+ S9 |
- S9 |
|
non-treated |
107 |
102 |
vehicle |
100 |
100 |
0.625 |
122 |
114 |
1.25 |
95 |
72 |
2.5 |
121 |
88 |
5 |
108 |
68 |
10 |
111 |
108 |
20 |
89 |
102 |
40 |
99 |
93 |
80 |
78 |
93 |
160 |
120 |
63 |
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
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