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EC number: 208-621-0 | CAS number: 535-87-5
- 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 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:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer reviewed publiication
Data source
Reference
- Reference Type:
- publication
- Title:
- Validation study of the in vitro micronucleus test in a Chinese hamster lung cell line (CHL/IU)
- Author:
- Taijiro Matsushima, Makoto Hayashi, Atsuko Matsuoka, Motoi Ishidate, Kunihiko F.Miura, Hidesuke Shimizu, Yuji Suzuki, Kanehisa Morimoto, Hiroko Ogura, Kanae Mure, Kimiko Koshi and Toshio Sofuni
- Year:
- 1 999
- Bibliographic source:
- Mutagenesis vol.14 no.6 pp.569–580, 1999
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- In vitro micronucleus test was performed to determine the toxic nature of 3,5-Diaminobenzoic acid
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian cell micronucleus test
Test material
- Reference substance name:
- 3,5-diaminobenzoic acid
- EC Number:
- 208-621-0
- EC Name:
- 3,5-diaminobenzoic acid
- Cas Number:
- 535-87-5
- Molecular formula:
- C7H8N2O2
- IUPAC Name:
- 3,5-diaminobenzoic acid
- Test material form:
- solid
Constituent 1
- Specific details on test material used for the study:
- - Name of test material: 3,5-diaminobenzoic acid
- EC name: 3,5-diaminobenzoic acid
- Molecular formula: C7H8N2O2
- Molecular weight: 152.1522 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data
Method
- Target gene:
- No data
Species / strain
- Species / strain / cell type:
- mammalian cell line, other: Chinese hamster lung cell line (CHL)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Eagle’s minimum essential medium supplemented with 10% heat inactivated (56°C for 30 min) calf serum
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: No data
- Periodically checked for karyotype stability: No data
- Periodically "cleansed" against high spontaneous background: No data - Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction prepared from phenobarbital- and 5,6-benzoflavone-pretreated male Sprague–Dawley rats was used
- Test concentrations with justification for top dose:
- 1500 – 3500 µg/mL (0, 1000, 1500, 2000, 2500, 3000 or 3500 µg/mL)
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
Cells seeded: 1X104–1X105 cells
DURATION
- Preincubation period: No data
- Exposure duration:
Without S9: 24, 48, 78 hrs, 6 hrs + 42 hrs (recovery) and 6 hrs + 66 hrs (recovery)
With S9: 6 hrs + 42 hrs (recovery) and 6 hrs + 66 hrs (recovery)
- Expression time (cells in growth medium): 66 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): 66 hrs
SELECTION AGENT (mutation assays): Acridine orange or Giemsa
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data
NUMBER OF REPLICATIONS: No data
NUMBER OF CELLS EVALUATED: The number of micronucleated cells per 1000 intact interphase cells was recorded.
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data
OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data
OTHER: No data - Rationale for test conditions:
- No data
- Evaluation criteria:
- The candidate MNs were categorized into three groups: very small (pin point) inclusions stained homogeneously (type 1) (those were not included for result evaluation), typical, i.e., smaller in diameter than ¼ of the normal main nucleus (type 2), and large, i.e., between ¼ and ½ the diameter of the normal main nucleus (type 3). The number of mitoses and abnormal cells (e.g., multinucleated cells, cells with abnormal nucleus) that appeared in the same microscopic field was also recorded.
As the negative control values ranged from 0.1 to 2.0% (Table II), chemicals yielding statistically significant MN frequencies with .4.0% were judged to be clear positives. When a compound induced a statistically significant MN frequency of < 4.0%, it was judged a weak positive. When no statistical significance was obtained, it was judged negative. - Statistics:
- The frequencies of cells with type 2 and/or type 3 MN in the treatment groups were compared with those of the concurrent negative control by Fisher’s exact test. The concentration–response relationship was evaluated by the Cochran– Armitage trend test. A result was considered statistically significant when the P-value of the Fisher’s exact test was smaller than 0.05 divided by the number of treatment groups and the P-value of the trend test was also smaller than 0.05.
Results and discussion
Test results
- Species / strain:
- mammalian cell line, other: Chinese hamster Lung Cell line (CHL)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not specified
- Additional information on results:
- No data
Applicant's summary and conclusion
- Conclusions:
- 3, 5 -Diaminobenzoic acid failed to induce mutation in Chinese hamster lung cell line (CHL) both with and without S9 metabolic activations system and hence is not likely to classify as a gene mutant in vitro.
- Executive summary:
In vitro micronucleus test was performed to determine the toxic nature of 3, 5 -Diaminobenzoic acid. The study was performed using Chinese hamster lung cell line (CHL) in the presence and absence of S9 metabolic activation system. The test chemical was used at concentration of 1500 – 3500µg/mL (0, 1000, 1500, 2000, 2500, 3000 or 3500µg/mL). The suration of exposure was 24, 48, 78 hrs, 6 hrs + 42 hrs (recovery) and 6 hrs + 66 hrs (recovery) without S9 and 6 hrs + 42 hrs (recovery) and 6 hrs + 66 hrs (recovery) with S9.The number of micronucleated cells per 1000 intact interphase cells was recorded. The candidate MNs were categorized into threegroups: very small (pin point) inclusions stained homogeneously (type 1) (those were not included for result evaluation), typical, i.e., smaller in diameter than ¼ of the normal main nucleus (type 2), and large, i.e., between ¼ and ½ the diameter of the normal main nucleus (type 3). The number of mitoses and abnormal cells (e.g., multinucleated cells, cells with abnormal nucleus) that appeared in the same microscopic field was also recorded. The statistically significant increase in MN frequency was obtained by short treatment (6166 h) with S9 mix, but it was not dose-dependent and the maximum frequency was only 1.7%, which was within the variations of the negative control value of the laboratory tested. The MN test showed clear negative results with 24, 48 and 72 h continuous treatments at up to 3500 mg/mL. 3, 5 -Diaminobenzoic acid failed to induce mutation in Chinese hamster lung cell line (CHL) both with and without S9 metabolic activations system and hence is not likely to classify as a gene mutant in vitro.
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