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EC number: 201-615-9 | CAS number: 85-56-3
- 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 bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From May 24, 1988 to June 20, 1988
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 988
- Report date:
- 1988
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 2-(4-chlorobenzoyl)benzoic acid
- EC Number:
- 201-615-9
- EC Name:
- 2-(4-chlorobenzoyl)benzoic acid
- Cas Number:
- 85-56-3
- Molecular formula:
- C14H9ClO3
- IUPAC Name:
- 2-(4-chlorobenzoyl)benzoic acid
- Test material form:
- solid
- Details on test material:
- Purity: 99.8 %
Constituent 1
Method
Species / strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- S-9 mix
- Test concentrations with justification for top dose:
- Experiment 1:
Doses: 0, 20, 100, 500, 2500 and 5000 ug/plate in DMS0 - 3 test plates per dose or per control
Experiment 2:
Doses: 0, 4, 20, 100, 500 and 2500 ug/plate in DMS0 - 3 test plates per dose or per control - Vehicle / solvent:
- Comlete solubility of the test substance in DMSO.
Controls
- Untreated negative controls:
- yes
- Remarks:
- vehicle: DMSO
- Positive controls:
- yes
- Positive control substance:
- other: 10 ug 2-aminoanthracene in DMSO (with S9), 5 ug N-methyl-N -nitro-N-nitrosoguanidine (MNNG) (without S-9))
- Details on test system and experimental conditions:
- S-9 mix:
The S-9 mix is prepared freshly prior to each experiment (1, 2). For this purpose, a sufficient amount of S-9 -fraction is thawed at room temperature and 3 volumes of S-9 fraction are mixed with 7 volumes of S-9 supplement (cofactors). This preparation, the so-called S-9 mix, is kept on ice until used. The concentrations of the cofactors in the S-9 mix are:
MgCl2 8mM
KCl 33 mM
glucose-6--phosphate 5 mM
NADP 4 mM
phosphate buffer (pH 7.4) 100 mM. The phosphate buffer is prepared by mixing an Na2HPO4 solution with an NaH2PO4 solution in a ratio of about 4:1.
Bacteria:
The indicator organisms TA 1535, TA 1537, TA 98 and TA 100 selected for this test are derivatives of Salmonella typhimurium. All strains have a defective excision repair System (uvr9), which prevents the repair of lesions which are induced in the DNA, and this deficiency results in greatly enhanced sensitivity of some mutagens. Furthermore, all strains show a considerably reduced hydrophilic polysaccharide layer (rfa), which leads to an increase in permeability to lipophilic substances.
The strains TA 1535 and TA 100 are derived from histidineprototrophic Salmonella strains by the substitution mutation his G 46 and are used to detect base pair substitutions. TA 1537 and TA 96 are strains for the detection of frameshift mutagens. These strains carry different frameshift markers, i.e. the .1 mutant his C 3076 in the case of TA 1537 and the .2 type his D 3052 in the case of TA 96.
The strains TA 96 and 1A 100 carry an R factor plasmid pKM 101 (4) and, in addition to having genes resistant to antibiotics, they have a modified postreplication DNA repair system, which increases the mutation rate by inducing a defective repair in the DNA; this again leads to a considerable increase in sensitivity.
Preincubation test
The experimental procedure is based on the method described by Yahagi et al. and Matsushime et al. 0.1 ml test solution, 0.1 ml bacterial suspension and 0.5 ml S-9 mix are incubated at 37 °C for the duration of 20 minutes. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the Vogel-Bonner agar plates within approx. 30 seconds.
Composition of the minimal glucose agar:
960 ml aqua dest.
20 ml Vogel-Bonner E medium
15 g Oifco bacto agar
20 g D-glucose, monohydrate.
After incubation at 37 °C for 48 hours in the dark, the bacterial colonies (his+ revertants) are counted.
Standard plate test
The experimental procedure is based on the method of Ames et al. Test tubes containing 2 ml portions of soft agar which consists of 100 mL agar (0.6 % agar + 0.6 % NaCl) and 10 mL amino acid solution (minimal amino acid solution for the determination of mutants: 0.5 mM histidine + 0.5 mM biotin) are kept in a water bath at 45°C, and the remaining components are added in the following order:
0.1 ml test solution
0.1 ml bacterial suspension
0.5 ml 5-9 mix (in tests with metabolic activation)
or
0.5 ml phosphate buffer (in tests without metabolic activation)
After mixing, the samples are poured onto Vogel-Bonner agar plates
Titer determination
Ttiter is determined only in the experiments with S-9 mix both without test substance (solvent only) and after adding the two highest amounts of substance. For this purpose, 0.1 mL of the overnight cultures is diluted to 10 –E06 in each case. Test tubes containing 2 ml portions of soft agar containing maximal amino-acid solution (5 mM histidine * 0.5 mM biotin) are kept in a water bath at 45 °C, and the remaining components are added in the following order:
0.1 ml solvent (without and with test substance)
0.1 ml bacterial Suspension (dilution: 10-E06)
0.5 ml S-9 mix
After mixing, the samples are poured onto the Vogel-8onner agar plates within approx. 30 seconds. After incubation at 37 °C for 48 hours in the dark, the bacterial colonies are counted.
Test design:
Experiment 1:
Strains: TA 1535, TA 100, TA 1537, TA 98
Doses: 0, 20, 100, 500, 2500 and 5000 ug/plate in DMS0
Type of test, Standard plate test with and without S-9 mix
Number of plates: 3 test plates per dose or per control
Experiment 2:
Strains: TA 1535, TA 100, TA 1537, TA 98
Doses: 0, 4, 20, 100, 500 and 2500 ug/plate in DMS0
Type of test, preincubation test with and without S-9 mix
Number of plates: 3 test plates per dose or per control - Evaluation criteria:
- Evaluation criteria
In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- doses >5000 µg/plate (SPT) or >2500 µg/plate (PIT)
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- doses >5000 µg/plate (SPT) or >2500 µg/plate (PIT)
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- doses >5000 µg/plate (SPT) or >2500 µg/plate (PIT)
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- doses >5000 µg/plate (SPT) or >2500 µg/plate (PIT)
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
Applicant's summary and conclusion
- Conclusions:
- Under the study conditions, the test substance is not mutagenic in the Salmonella typhimurium strains TA 1535, TA 100, TA 1537, TA 98 at the dose range of: 20 µg - 5000 µg/plate (SPT) and at the dose range of: 4 µg - 2500 µg/plate (PIT) in the absence and presence of metabolic activation.
- Executive summary:
A study was conducted to determine the mutagenic potential of the test substance according to OECD Guideline 471 in Bacterial Reverse Mutation Test. The test substance was examined using four strains of Salmonella typhimurium (TA 1535, TA 100, TA 1537, and TA 98). The test was performed in two experiments in the presence and absence of S9-mix (rat liver S9-mix induced by Aroclor 1254). DMSO was used as a vehicle. Toxic effects were monitored at doses: 20 µg - 5000 µg/plate in the Standard plate test (SPT) and 4 µg - 2500 µg/plate in the Preincubation test (PIT). In both experiments 3 test plates per dose or per control were investigated. Bacteriotoxic effect were observed at doses >5000 µg/plate (SPT) or >2500 µg/plate (PIT). An increase in the number of his revertants was not observed both in the SPT and in the PIT either without S-9 mix or after the addition of a metabolizing system. Under the study conditions, the test substance is not mutagenic in the Salmonella typhimurium strains TA 1535, TA 100, TA 1537, TA 98 at the dose range of: 20 µg - 5000 µg/plate (SPT) and at the dose range of: 4 µg - 2500 µg/plate (PIT) in the absence and presence of metabolic activation (Gelbke, 1988).
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