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EC number: 201-474-3 | CAS number: 83-41-0
- 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:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- restrictions: no historical control data; only 100 metaphases analyzed; no statistical evaluation; methods used for determination of cytotoxicity no longer recommended (PE assay) or not recommended for cell lines (mitotic index)
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 988
- Report date:
- 1988
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
Test material
- Reference substance name:
- 3-nitro-o-xylene
- EC Number:
- 201-474-3
- EC Name:
- 3-nitro-o-xylene
- Cas Number:
- 83-41-0
- Molecular formula:
- C8H9NO2
- IUPAC Name:
- 1,2-dimethyl-3-nitrobenzene
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
Charge 22
- Purity test date:
12.10.1987
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
4°C; light protected
- Solubility and stability of the test substance in the solvent/vehicle:
> 3 months in H20 and DMSO
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
On the day of the experiment, the test article was dissolved in ethanol. The final
concentration of ethanol in the culture medium did not exceed 1 % v/v.
Method
Species / strain
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix according to Ames et al.
- Test concentrations with justification for top dose:
- without S9 mix:
7 h: 25; 50; 100; 150 µg/ml
18 h: 15; 100; 150; 200; 250**; 300** µg/ml
28 h: 150; 200; 250**; 300** µg/ml
with S9 mix:
7 h: 25; 50; 100; 150 µg/ml
18 h: 15; 100; 150; 200; 250**; 300** µg/ml
28 h: 150; 200; 250**; 300** µg/ml
**slight precipitation in the culture medium
These concentration ranges have been determined in a pre-experiment using the plating efficiency assay.
Treatment of the cells with 200 µg/ml in the presence of S9 mix reduced clearly the plating efficiency of the V79 cells. Higher concentration precipitated in the culture medium. Also the mitotic index was reduced after treatment with 150 and 200 µg/ml at fixation interval 18 h with and without S9 mix.
The following dose levels were were selected to evaluate metaphases for cytogenetic damage:
without S9 mix:
7 h: 100 µg/ml
18 h: 15; 100; 150 µg/ml
28 h: 150 µg/ml
with S9 mix:
7 h: 150 µg/ml
18 h: 15; 150; 200 µg/ml
28 h: 150 µg/ml
In the main-experiment in the absence of S9 mix at fixation intervals 7, 18 and 28 h 100 and 150 µg/ml were chosen as maxima concentrations because with higher concentrations not enough scorable cells could be found.
In the presence of S9 mix at fixation intervals 7 and 28 h cells after treatment with 150 µg/ml as maximum concentration could be scored for cytogenetic damage; at interval 18 h cells after treatment with 200 µg/ml could be evaluated as highest concentration. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used:
ethanol
- Justification for choice of solvent/vehicle:
The solvent was chosen according to its solubility properties and its relative non-toxicity for the cells.
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION:
in medium
DURATION
- Exposure duration: 4h
- Fixation time (start of exposure up to fixation or harvest of cells):
Preparation of chromosomes was done 7 h (high dose), 18 h (low,medium and high dose), and 28 h (high dose) after start of treatment with the test article.
SPINDLE INHIBITOR (cytogenetic assays):
colcemid
STAIN (for cytogenetic assays):
V79
NUMBER OF REPLICATIONS:
2 per experiment in 2 experiments (I and II)
METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED:
after treatment with colcemid, the cells were treated on the slides in the chambers with hypotonic solution for 20 min at 37 °C. After incubation in the hypotonic solution the cells are fixed with 3:1 absolute methanol:glacial acetic acid. After fixation the cells were stained with aceto-orcein.
NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells):
At least 100 well spread metaphases per slide
DETERMINATION OF CYTOTOXICITY
- Method: plating efficiency index (colony forming ability); mitotic index - Evaluation criteria:
- A test article is considered mutagenic in this assay if it induces either a significant dose-related increase in the number of structural chromosomal aberrations or a significant and reproducible positive response for at least one of the test points.
A test article which producing neither a significant dose-related increase in the number of structural chromosomal aberrations nor a reproducible positive response at any of the test points is considered non-mutagenic in this assay. - Statistics:
- According to the study report, a statistical evaluation of the results could not be performed because there was no adequate statistical procedure available.
Results and discussion
Test results
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
Applicant's summary and conclusion
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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