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EC number: 202-764-2 | CAS number: 99-54-7
- 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 gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Study well documented, meets generally accepted scientific priciples, acceptable for assessment, test substance techn. grade
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 986
- Report date:
- 1986
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- -addition of different concentrations of the metabolic activation system
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Reference substance name:
- 1,2-dichloro-4-nitrobenzene
- EC Number:
- 202-764-2
- EC Name:
- 1,2-dichloro-4-nitrobenzene
- Cas Number:
- 99-54-7
- Molecular formula:
- C6H3Cl2NO2
- IUPAC Name:
- 1,2-dichloro-4-nitrobenzene
- Reference substance name:
- 1,2-dichloro-3-nitrobenzene
- EC Number:
- 221-717-7
- EC Name:
- 1,2-dichloro-3-nitrobenzene
- Cas Number:
- 3209-22-1
- Molecular formula:
- C6H3Cl2NO2
- IUPAC Name:
- 1,2-dichloro-3-nitrobenzene
Constituent 1
Constituent 2
Method
- Target gene:
- HGPRT
Species / strain
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Remarks:
- CHO-K1BH4
- Details on mammalian cell type (if applicable):
- CELLS USED
- Type and source of cells: CHO-K1-BH4, Dr. Abraham W. Hsie, Biology Division, Oak Ridge National Laboratories, P.O. Box Y, Oak Ridge, Tennessee 37380
- Suitability of cells: Chemicals capable of inducing mutations have been shown to incraase the
forward mutation frequency at the hgprt loclus in Chinese Hamster Ovary Cells
The stock cultures of CHO-K1-BH4 cell line are maintained in frozen aliquots in a Revco Ultra-low Freezer. Cultures of CHO-K1-BH4 cell line were prepared from stock cultures known to have a stable spontaneous mutation frequency of 0 - 10 x 10E-6 mutants per cell, however, values up to 20 x 10E-6 were deemed acceptable.
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver S9-mix: 1 %, 2 %, 5%, 10%
- Test concentrations with justification for top dose:
- 1st experiment: -S9-mix: 100, 120, 150 µg/mL, +S9-mix (1, 2, 5, 10%): 50, 150, 200 µg/mL
2nd experiment: 0, 25, 50, 125, 200, 250 µg/mL (cytotoxicity of approximately 92, 70, 43, 30 and 15 % mean relative survival without metabolic activation and 81, 71, 54, 42 and 7 % mean relative survival with metabolic activation, respectively.) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- N-dimethylnitrosamine
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- NUMBER OF REPLICATIONS:
- Number of cultures per concentration: duplicate (1st experiment), triplicate (2nd experiment)
- Number of independent experiments: 2
METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): 5 x 10E5 cells in 5 ml of medium
- Test substance added in medium
TREATMENT AND HARVEST SCHEDULE:
- Exposure duration/duration of treatment: 5 h
- Harvest time after the end of treatment (sampling/recovery times): 19 h
FOR GENE MUTATION:
- Expression time (cells in growth medium between treatment and selection): 7 - 8 d
- Selection time (if incubation with a selective agent): 7 d
- Method used: agar or microwell plates for the mouse lymphoma assay.
- If a selective agent is used (e.g., 6-thioguanine or trifluorothymidine), indicate its identity, its concentration and, duration and period of cell exposure: For mutant selection, 6.25 mL of 10E-3 M 6-thioguanine solution was added to 494 mL of hypoxanthine free medium. To each of 5-100 mm plates 8 mL of the 6-TG medium were added and 2 mL of the 1 x 10 E5 cells/mL aliquot, for a total of 2 x 10E5 cells/plate. The plates were incubated for 7 days at 37°C in 5 % CO2 in air at 90+ % humidity to allow for colony formation.
- Number of cells seeded and method to enumerate numbers of viable and mutants cells:
METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method, e.g.: relative survival (RS), cloning efficiency - Evaluation criteria:
- positive: mutation frequency significant greater than control, in one concentration and mean survival of at least 10 %, dose response relationship
negative: none mutation frequency greater than the of the solvent control, no dose-response relationship - Statistics:
- one-way analysis of variance method outlined by Snee and Irr (1981) one-tailed student's t-test using pooled , intergroup variance; dose-response relationship
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:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RANGE-FINDING/SCREENING STUDIES (if applicable):
1. toxicity test prior to testing:
dose levels:
0.33, 1.0, 3.3, 10, 33.3, 100, 333, 1000 µg/ml in the presence of 0, 1, 2, 5, and 10 % Ariclor1254 induced rat liver S9-mix
incubation time: 5 hours
result:
1000 µg/ml: cytotoxicity at all concentrations of S9-mix
333 µg/ml: reduced relative cell survival (0-10% S9): 36%, 36% 34%, 38%, 18%
2. preliminary mutagenicity screen:
dose levels:
-S9-mix: 100, 120, 150 µg/ml
+S9-mix (1, 2, 5, 10%): 50, 150, 200 µg/ml
incubation time: 5 hours with TS and after removal of TS for 19 hours and after washing for additional 7 days
result:
survival:
-S9-mix: 99, 94, 47 %
+S9-mix: 1%: 79/45/10% survival; 2%: 86/48/25% survival; 5%: 91/73/ 42% survival; 10%: 99/80/78% survival
there were no significant increases in the mutation levels when compared to the negative controls
STUDY RESULTS
- Concurrent vehicle negative and positive control data:
Mean mutation frequency (with S9-mix - without S9-mix):
-negative controls:
untreated controls: 0.6 - 1.9
DMSO- control: 0.8 - 0.6
-positive controls:
EMS: 275 (without S9-mix)
DMN: 265 (with S9-mix)
Gene mutation tests in mammalian cells:
- Results from cytotoxicity measurements:
o Relative total growth (RTG) or relative survival (RS) and cloning efficiency:
relative cell survival (low to high dose):
-S9-mix: 92, 70, 43, 30, 15 %
+S9-mix: 81, 71, 54, 42, 7 %
mean mutation frequency (low to high dose: with/without S9-mix):
0.0/1.4, 1.3/1.1, 0.8/0.5, 0.6/1.5, 1.0/1.4
->no statistically significant difference when compared to the negative controls
Applicant's summary and conclusion
- Conclusions:
- 1,2 -dichloro-4 -nitrobenzene technical grade was tested negative in this CHO/HPRT Mammalian cell forward gene mutation assay.
- Executive summary:
1,2 -dichloro-4 -nitrobenzene technical grade was tested in the CHO/HPRT Mammalian cell forward gene mutation assay and yielded negative result (Monsanto Co 1986).
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