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EC number: 201-089-0 | CAS number: 78-16-0
- 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
- 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
- Study period:
- 23 Feb - 26 Jul 2000
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Guideline study with acceptable restrictions (no details on analytical purity of the test substance given).
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 000
- Report date:
- 2000
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no details on analytical purity of the test substance given
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no details on analytical purity of the test substance given
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test
- Deviations:
- yes
- Remarks:
- no details on analytical purity of the test substance given
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- 189120-64-7
- Cas Number:
- 189120-64-7
- IUPAC Name:
- 189120-64-7
- Details on test material:
- - Name of test material (as cited in study report): MRD-99-429
- Physical state: pale yellow liquid
- Analytical purity: no data
- Expiration date of the lot/batch: 09/2004 (container 1) and 03/2005 (container 2)
- Storage condition of test material: at room temperature
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: in the absence of S9 mix: McCoy's 5A Medium containing 10% foetal bovine serum and 2 mM L-glutamine; in the presence of S9 mix: serum-free McCoy's 5A Medium containing 2 mM L-glutamine
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Metabolic activation:
- with and without
- Metabolic activation system:
- co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of Sprague Dawley rats treated with Aroclor
- Test concentrations with justification for top dose:
- Range-finder toxicity test: 0, 20, 39, 78, 156, 313, 625, 1250 and 2500 µg/mL (3 h treatment), with and without S9
Initial and repeat experiment (main assay): 25, 75, 250, 750 and 2500 µg/mL (3 h treatment), with and without S9
Chromosome aberration analysis: 75, 750, and 2500 µg/mL (without S9); 25, 250 and 2500 µg/mL (with S9) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: a vehicle solubility test was previously performed to assess the solubility of the test substance in DMSO, water and acetone. The results of this test showed that the test substance was soluble in acetone at the concentrations required for this study. The non-cytotoxic dose volume for acetone is 50 µL/flask, therefore based on solubility, 2500 µg/mL was the highest dose that could be achieved in this assay.
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- acetone
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- - S): 9,10-dimethyl- 1,2-benzanthracene (DMBA, 10 µg/mL in acetone); + S9: 1-methyl-3-nitro-1-nitrosoguanidine (MNNG, 0.6 µg/mL in acetone
- Positive control substance:
- 9,10-dimethylbenzanthracene
- other: 1-methyl-3-nitro-1-nitrosoguanidine
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 3 h
- Fixation time (start of exposure up to fixation or harvest of cells):
A) Range-finder toxicity test: 19 h
B) Initial experiment (main assay): 19 h
C) Repeat experiment (main assay): 19 and 43 h
SPINDLE INHIBITOR (cytogenetic assays): 0.2 µg/mL Colcemid®
STAIN (for cytogenetic assays): 5% Giemsa
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index and cell confluency (range-finder toxicity test and main assay); other: cell count (range-finder toxicity test) - Evaluation criteria:
- For a test substance to be considered as positive, one of the following conditions must be met:
- a statistically significant dose-related increase in the mean percentage of aberrant cells and, in at least one of the treatment groups, the mean percentage of aberrant cells exceeds 5%.
OR
- 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 the test substance induces chromosomal aberrations in cultured mammalian somatic cells. If neither of the above conditions are met, the test substance is considered as non-mutagenic in this system. - Statistics:
- The number of cells with at least one aberrant chromosome and the number of cells examined in each replicate were used for statistical analysis. The number of aberrant individual chromosomes per cell was not statistically analysed.
The results of the positive control group were compared to the appropriate vehicle control group by the Fisher Exact Test to assure that the assay was performed in an appropriate manner.
To test for homogeneity of the replicates, each pair of replicates was compared by Fisher Exact Test. Two times the sum of the log of the individual two-sided significance levels was compared to chi-square distribution with 2k degrees of freedom (k is the number of replicate pairs). If the test failed, further investigation would be pursued and the remaining analyses would not be performed.
To test for differences among the control and the treated groups, a 2x2-Fisher Exact Test was performed. If differences were shown to exist at the 0.05 level or less, individual 2x2-Fisher Tests were performed to determine which of the treated groups differed from the control group. A permutation test (Hoeffding, 1952) was performed to test for dose-related trends. Statistical analysis included the calculation of means for percent confluency, percent mitotic cells, percent aberrant cells, percent frequency of aberrations, and cell counts. Significance was reported when p < 0.05.
Results and discussion
Test results
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RANGE-FINDING/SCREENING STUDIES: in the range-finder toxicity test, visual solubility observations were made 3 h after treatment. Cloudiness was evident at all concentrations with S9. Complete solubility was observed at concentrations ranging from 10 to 39 µg/mL without S9. Cloudiness was evident at concentrations of 78 µg/mL through 625 µg/mL without S9. Oil droplets were noted without metabolic activation at concentrations ≥ 1250 µg/mL. A notable decrease in confluency (≥ 50% reduction compared with vehicle controls) were observed at concentrations of 625 µg/mL and 1250 µg/mL with S9 and 2500 µg/mL without S9. Decreases in cell survival and mitotic index (MI) of ≥ 50% from the vehicle control were not observed in either the activated or non-activated series. Cell morphology was normal at all concentrations tested, however debris of undetermined origin was noted in activated flasks at concentrations ≥ 156 µg/mL and non-activated flasks ≥ 313 µg/mL. Based on these results, 25, 75, 250, 750 and 2500 µg/mL were selected as concentrations for the main chromosomal aberration assay.
ADDITIONAL INFORMATION ON CYTOTOXICITY: in both experiments of the main study, there was no notable decrease in the percent confluency (≥ 50% reduction compared to the vehicle control) at any concentration with or without S9. Cell morphology was normal at all concentrations tested. Similarly, no notable decrease (≥ 50%) in MI values was noted for the test substance concentrations. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
CHROMOSOME ANALYSIS
A) INITIAL ASSAY
There were no statistically significant differences or dose-related trends in the percentage of cells with chromosomal aberrations between the treated and the control groups either with or without metabolic activation. The percentage of aberrant cells in the vehicle control groups, with and without metabolic activation, was 2.5 and 1.0%, respectively. Both values fell within the 0 to 5% range of acceptance for the vehicle control. The percentage of aberrant cells in the treatment groups ranged from 2.0 to 3.0% for the metabolically activated series and from 0.5 to 3.0% in the non-activated series.
Table 1. Test results of the initial experiment
Test item |
Concentration in µg/mL |
Mitotix index in % of vehicle control |
Aberrant cells in % per 200 cells scored |
Frequency of abberations in % per 200 cells scored |
|
with gaps |
without gaps |
||||
Exposure period 3 h, fixation time 19 h, without S9 mix |
|||||
Acetone |
a |
100 |
1.0 |
1.5 |
1 |
MNNG |
0.6 |
26 |
12.0** |
5.0 |
12.5 |
Test substance |
75 |
77 |
0.5 |
2.5 |
0.5 |
750 |
80 |
3.0 |
1.0 |
3 |
|
2500 |
54 |
2.0 |
0.0 |
2 |
|
Exposure period 3h, fixation time 19 h, with S9 mix |
|||||
Acetone |
a |
100 |
2.5 |
2.5 |
3.0 |
DMBA |
10 |
59 |
19.0** |
5.0 |
26.0 |
Test substance |
25 |
112 |
3.0 |
3.0 |
3.0 |
250 |
100 |
2.5 |
1.5 |
2.5 |
|
2500 |
104 |
2.0 |
3.0 |
2.0 |
MNNG = 1-methyl-3-nitro-1-nitrosoguanidine; DMBA = 9,10-dimethyl-1,2-benzanthracene
a = concentration: 50 µL/flask; b = positive controls were not required for the 43 h harvest
*p < 0.05; **p < 0.01
B) REPEAT ASSAY
There was a statistically significant difference (p < 0.05) in the percentage of cells with chromosomal aberrations between the vehicle and the highest concentration (2500 µg/mL) in the 19 h harvest with metabolic activation. The permutation test for the 19 h harvest with metabolic activation also appears to be significant (p<0.01) indicating a dose-related trend in the percentage of cells with chromosomal aberrations. This increase in the percentage of aberrant cells does not appear to be biologically significant due to the fact that the percentage of aberrant cells fell within the acceptable range for the vehicle control (0-5%).
The percentage of aberrant cells in the vehicle control groups at both harvest intervals (with and without metabolic activation) was 2.5% or less. These values fell within the 0 to 5% range of acceptance for the vehicle control. The percentage of aberrant cells observed in the 19 h treated groups ranged from 1 to 3.0%. The percentage of aberrant cells observed in the 43 h treated groups ranged from 0.5% to 2.5%.Table 2. Test results of the repeat experiment
Test item |
Concentration in µg/mL |
Mitotix index in % of vehicle control |
Aberrant cells in % per 200 cells scored |
Frequency of abberations in % per 200 cells scored |
|
with gaps |
without gaps |
||||
Exposure period 3 h, fixation time 19 h, without S9 mix |
|||||
Acetone |
a |
100 |
1.0 |
1.0 |
1.0 |
MNNG |
0.6 |
42 |
8.0** |
2.0 |
13.5 |
Test substance |
75 |
89 |
2.0 |
2.5 |
2.5 |
750 |
65 |
1.5 |
1.0 |
1.5 |
|
2500 |
82 |
1.5 |
1.0 |
2.0 |
|
Exposure period 3 h, fixation time 43 h, without S9 mix |
|||||
Acetone |
a |
100 |
2.5 |
2.5 |
2.5 |
MNNG |
0.6 |
b |
b |
b |
b |
Test substance |
75 |
105 |
0.5 |
1.0 |
0.5 |
750 |
95 |
0.5 |
1.5 |
0.5 |
|
2500 |
68 |
1.5 |
2.0 |
1.5 |
|
Exposure period 3 h, fixation time 19 h, with S9 mix |
|||||
Acetone |
a |
100 |
0.0 |
1.5 |
0.0 |
DMBA |
10 |
66 |
10.5** |
7.0 |
11.5 |
Test substance |
25 |
107 |
1.0 |
1.0 |
1.0 |
250 |
105 |
1.0 |
1.0 |
1.0 |
|
2500 |
60 |
3.0* # |
1.0 |
3.5 |
|
Exposure period 3 h, fixation time 43 h, with S9 mix |
|||||
Acetone |
a |
100 |
1.0 |
1.0 |
1.0 |
DMBA |
0.6 |
b |
b |
b |
b |
Test substance |
25 |
89 |
2.5 |
2.5 |
2.5 |
250 |
70 |
1.5 |
1.0 |
1.5 |
|
2500 |
68 |
2.0 |
0.0 |
2.0 |
MNNG = 1-methyl-3-nitro-1-nitrosoguanidine; DMBA = 9,10-dimethyl-1,2-benzanthracene
a = concentration: 50 µL/flask; b = positive controls were not required for the 43 h harvest
*p < 0.05; **p < 0.01
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
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