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EC number: 308-067-0 | CAS number: 97862-23-2
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Ames assay:
The test chemical did not induce mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian chromosome aberration study:
The test chemical did not induce chromosome aberration in Chinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian cell gene mutation assay:
The test chemical did not induce gene mutation in mammalian cell line in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Remarks:
- Experimental data from various test chemicals
- Justification for type of information:
- Data for the target chemical is summarized based on the data from various test chemicals
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- WoE derived based on the experimental data from various test chemicals
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Histidine
- Species / strain / cell type:
- S. typhimurium, other: TA1535, TA100, TA1537, TA1538, TA1978 and TA98
- Remarks:
- 1
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- other:
- Species / strain / cell type:
- S. typhimurium, other: TA97, TA98, TA100
- Remarks:
- 2
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- Strains TA1535, TA1537 and TA1538 have mutations hisG46, hisC3076 and hisD3052 in their respective histidine operons, resulting in a nutritional requirement for histidine. In addition they possess deletions in the uvr region of the genome removing their capacity for excision repair and contain rfa (deep rough) mutations resulting in the impaired synthesis of the polysaccharide side chains of their surface lipopolysaccharide. Strain TA1978 resembles TA1538 except that it possesses the normal uvr repair system and a mutation in gal E. Strains TA100 and TA98 are derived from TA1535 and TA1538 respectively by insertion of the R factor plasmid pKM101.
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254 induced S9 fraction was obtained from Female albino rats
- Test concentrations with justification for top dose:
- 1. 100-200 µg/plate
2. 0, 5, 10, 50 or 200 µg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- For TA1535 / 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- For TA100 / 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- For TA1537 / 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- Remarks:
- TA1538 and TA98. / 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-anthramine
- Remarks:
- Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA1978 and TA98 / 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Remarks:
- 2
- Details on test system and experimental conditions:
- 1. METHOD OF APPLICATION: as impregnation on paper disk
DURATION
- Preincubation period: No data
- Exposure duration: 72 hrs
- Expression time (cells in growth medium): 72 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data
NUMBER OF REPLICATIONS: ): Duplicate
NUMBER OF CELLS EVALUATED: No data
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
2. METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period:No data
- Exposure duration:2 days (48 hrs)
- Expression time (cells in growth medium):2 days (48 hrs)
- Selection time (if incubation with a selection agent):No data
- Fixation time (start of exposure up to fixation or harvest of cells):No data
SELECTION AGENT (mutation assays):No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays):No data
NUMBER OF REPLICATIONS:Duplicate
NUMBER OF CELLS EVALUATED:No data
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:Yes, cell growth was noted
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:
- 1. Growth inhibition was noted
2. The plates were observed for a dose dependent increase in the number of revertants/plate - Statistics:
- No data
- Species / strain:
- S. typhimurium, other: TA1535, TA100, TA1537, TA1538, TA1978 and TA98
- Remarks:
- 1
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium, other: TA97, TA98, TA100
- Remarks:
- 2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Additional information on results:
- No data
- Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
- Executive summary:
Data available for the various test chemicals was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by spot test using Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA1978 and TA98 with and without S9 metabolic activation system. 100-200 µg of the test material dissolved in 10-20µL DMSO/disc was applied to 6mm paper concentration discs and used for mutagenicity testing. From 1 to 4 discs were applied per plate. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did not induce mutation in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA1978 and TA98 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium strains TA97, TA98, TA100 with and without PCB induced S9 metabolic activation system at dose levels of 0, 5, 10, 50 or 200µg/plate. The plates were incubated for 48 hrs and the number of dose dependent increase in the revertants was counted. The test chemical did not induce reversion of histidine gene mutation in Salmonella typhimurium strains TA97, TA98, TA100 both in the presence and absence of PCB induced rat liver S9 fraction and hence it is not likely to classify as a gene mutant in vitro.
Based on the observations made, the test chemical did not induce mutation in Salmonella typhimurium strains in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Remarks:
- Experimental data from various test chemicals
- Justification for type of information:
- Data for the target chemical is summarized based on the data from various test chemicals
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- WoE derived based on the experimental data from various test chemicals
- GLP compliance:
- not specified
- Type of assay:
- other: In vitro mammalian chromosome aberration study
- Target gene:
- 1. No data
- Species / strain / cell type:
- mammalian cell line, other: Chinese hamster cultured cells (CHL/IU)
- Remarks:
- 1
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Eagle MEM culture medium supplemented with 10% fetal bovine serum
- Properly maintained: No data
- 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
- Species / strain / cell type:
- mammalian cell line, other: Chinese hamster cultured cells (CHL/IU)
- Remarks:
- 2
- Details on mammalian cell type (if applicable):
- No data
- Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- 1. Continuous treatment: 0, 0.3, 0.7, 1.3 µg/mL
Short term treatment method: 0, 06, 1.1, 2.2 µg/mL
2. Continuous treatment: 0, 19.1, 38.3, 76.6, 153, 306, 613, 1230, 2450 µg/mL
Short term treatment method:
With S9: 0, 38.3, 76.6, 153, 306, 613, 1230, 2450 µg/mL
Without S9: 0, 19.1, 38.3, 76.6, 153, 306, 613, 1230, 2450 µg/mL - Vehicle / solvent:
- 1. - Vehicle(s)/solvent(s) used: 0.5% CMC Na
- Justification for choice of solvent/vehicle: The test chemical was soluble in CMC Na
2. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- CMC Na
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Remarks:
- 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Remarks:
- 2
- Details on test system and experimental conditions:
- 1. METHOD OF APPLICATION: in medium
Cells at the start of the experiment: 20000 cells
DURATION
- Preincubation period: No data
- Exposure duration: Direct method: 24 and 48 hrs
Short term treatment method with S9: 6 hrs
- Expression time (cells in growth medium):
Direct method: 24 and 48 hrs
Short term treatment method with S9: 18 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: No data
NUMBER OF CELLS EVALUATED: For structural abnormalities, 200 metaphase cells per group and 800 division metastatic cells for multiplicative cells were analyzed.
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
2. METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data
- Exposure duration: Direct method: 24 and 48 hrs
Short term treatment method with S9: 6 hrs
- Expression time (cells in growth medium):
Direct method: 24 and 48 hrs
Short term treatment method with S9: 18 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: No data
NUMBER OF CELLS EVALUATED: For structural abnormalities, 200 metaphase cells per group were analyzed.
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 presence or absence of structural abnormality such as chromosome type or chromosome type gap, The presence or absence of cells (polyploid) was also observed.
- Statistics:
- 1. A significant difference test (p <0.05) between the negative control group and the test substance treated group and between the negative control group and the positive control group was performed on the occurrence frequency of cells having chromosomal abnormality by Fisher's exact probability test method. According to the judgment criteria of Ishikan et al, the frequency of cells with chromosomal abnormality is negative, negative 5% or higher and less than 10% positive false positive 10% or higher Respectively.
2. No data - Species / strain:
- mammalian cell line, other: Chinese hamster cultured cells (CHL/IU)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- 1. TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data
RANGE-FINDING/SCREENING STUDIES: To determine the treatment concentration of the test substance used for the chromosomal aberration test, the influence of the test substance on cell proliferation was investigated. The growth inhibitory action of the test substance on CHL / IU cells was determined by measuring the proliferation degree of each group using a monolayer culture cell densitometer, and the negative control group was used as an index.
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data
2. No data - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce chromosome aberration in Chinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
- Executive summary:
Data available for the test chemicals was reviewed to determine the mutagenic nature. The studies are mentioned below:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using Chinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in 0.5% CMC Na and used at dose level of 0, 0.3, 0.7, 1.3 µg/mL in continuous treatment method and 0, 06, 1.1, 2.2 µg/mL in short term treatment method. The doses for the main study were based on data from preliminary dose range finding study. Concurrent solvent and positive control plates were also included in the study. The cells were exposed to the test chemical for 24 or 48 hrs in the continuous treatment method and for 6 hrs in the short term treatment method. Two hours before the end of the culture, colcemid was added to the culture solution to a final concentration of about 0.1 μg / ml. Chromosome specimens were prepared according to a conventional method. Giemsa stained six slide specimens were prepared for each petri dish. The presence or absence of structural abnormality such as chromosome type or chromosome type gap, the presence or absence of cells (polyploid) was also observed.The test chemical did not induce chromosome aberration inChinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using Chinese hamster cultured cells (CHL) in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 19.1, 38.3, 76.6, 153, 306, 613, 1230, 2450µg/mL without S9 and 0, 38.3, 76.6, 153, 306, 613, 1230, 2450µg/mL with S9 in the short term treatment method and 0, 19.1, 38.3, 76.6, 153, 306, 613, 1230, 2450µg/mL in continuous treatment method. The doses for the main study were based on data from preliminary dose range finding study. Concurrent solvent and positive control plates were also included in the study. The cells were exposed to the test chemical for 24 or 48 hrs in the continuous treatment method and for 6 hrs in the short term treatment method.The test chemical did not induce chromosome aberration inChinese hamster cultured cells (CHL) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Based on the observations made, the test chemical did not induce chromosome aberration in Chinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Remarks:
- Experimental data from various test chemicals
- Justification for type of information:
- Data for the target chemical is summarized based on the from various test chemicals
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- WoE derived based on the experimental data from various test chemicals
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian cell gene mutation tests using the thymidine kinase gene
- Target gene:
- 1. Thymidine kinase
2. HGPRT locus - Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Remarks:
- 1
- Details on mammalian cell type (if applicable):
- No data
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Remarks:
- 2
- Details on mammalian cell type (if applicable):
- No data
- Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- Liver S9 fraction from phenobarbital/β-naphthoflavone induced rats was used as exogenous metabolic activation system
- Test concentrations with justification for top dose:
- 1. Experiment I: 0, 1.6, 3.3, 6.5, 13.0 and 26.0 μg/ml without and with S9-mix
Experiment II: 0, 1.6, 3.3, 6.5, 13.0 and 26.0 μg/ml without S9-mix
2. No data - Vehicle / solvent:
- 1. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO
2. No data - Untreated negative controls:
- yes
- Remarks:
- As per OECD guideline
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: As per OECD guideline
- Remarks:
- 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Remarks:
- 2
- Details on test system and experimental conditions:
- 1. METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data
- Exposure duration: Experiment 1: 4 hrs
Experiment 2: 24 hrs
- Expression time (cells in growth medium):
Experiment 1: 72 hrs
Experiment 2: 48 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: No data
NUMBER OF CELLS EVALUATED: No data
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Toxicity was measured in the main experiments as percentage relative total growth of the treated cultures relative to the total growth of the solvent control cultures.
OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data
OTHER: No data
2. METHOD OF APPLICATION: in medium - Rationale for test conditions:
- No data
- Evaluation criteria:
- The cell line was observed for gene mutation at the respective locus
- Statistics:
- No data
- Species / strain:
- mouse lymphoma L5178Y cells
- Remarks:
- 1
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Remarks:
- 2
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Additional information on results:
- 1. CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: Yes
- Other confounding effects: No data
RANGE-FINDING/SCREENING STUDIES: Test concentrations were based on the results of a pre-test on toxicity measuring relative suspension growth.
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data
2. No data - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce gene mutation in mammalian cell line in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
- Executive summary:
Data available for the various test chemicals was reviewed to determine the mutagenic nature. The studies are as mentioned below:
In vitro mammalian cell gene mutation assay was performed to determine the mutagenic nature of the test chemical. The study was performed using L5178Y tk+/- mouse lymphoma cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level 0, 1.6, 3.3, 6.5, 13.0 and 26.0 μg/ml in experiment 1 (with and without S9) and in experiment 2 (without S9). The doses for the main study were dependent on the dose range finding study conducted. At the end of the treatment at 13 and 26 μg/ml in all cultures of both main experiments precipitation was observed by the unaided eye. No substantial and reproducible increase of the mutant frequency was observed in both experiments. A minor increase exceeding the threshold of twice the mutant frequency of the corresponding solvent control was observed in the second culture of the second experiment. Since the historical range of negative and solvent controls was not exceeded, the effect occurred at precipitating concentrations and was not reproducible in the other culture it was considered an artificial effect due to the precipitation of the test item rather than indicating a possible mutagenic effect. Based on the observations made, thetest chemical did not induce gene mutation in L5178Y tk+/- mouse lymphoma cells in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian cell gene mutation assay was performed to determine the mutagenic nature of the test chemical. The study was performed using V79 cells with and without co-cultivation with rat hepatocytes. A test is considered to be positive if a doubling of the background mutation frequency and/or a dose dependency is observed. Based on the observations made, thetest chemical did not induce gene mutation inV79 cells and hence it is not likely to classify as a gene mutant in vitro.
Based on the observations made, the test chemical did not induce gene mutation in mammalian cell line in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
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Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Data available for the various test chemicals was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
Ames assay:
Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed by spot test using Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA1978 and TA98 with and without S9 metabolic activation system. 100-200 µg of the test material dissolved in 10-20µL DMSO/disc was applied to 6mm paper concentration discs and used for mutagenicity testing. From 1 to 4 discs were applied per plate. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did not induce mutation in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA1978 and TA98 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium strains TA97, TA98, TA100 with and without PCB induced S9 metabolic activation system at dose levels of 0, 5, 10, 50 or 200µg/plate. The plates were incubated for 48 hrs and the number of dose dependent increase in the revertants was counted. The test chemical did not induce reversion of histidine gene mutation in Salmonella typhimurium strains TA97, TA98, TA100 both in the presence and absence of PCB induced rat liver S9 fraction and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian chromosome aberration study:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using Chinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in 0.5% CMC Na and used at dose level of 0, 0.3, 0.7, 1.3 µg/mL in continuous treatment method and 0, 06, 1.1, 2.2 µg/mL in short term treatment method. The doses for the main study were based on data from preliminary dose range finding study. Concurrent solvent and positive control plates were also included in the study. The cells were exposed to the test chemical for 24 or 48 hrs in the continuous treatment method and for 6 hrs in the short term treatment method. Two hours before the end of the culture, colcemid was added to the culture solution to a final concentration of about 0.1 μg / ml. Chromosome specimens were prepared according to a conventional method. Giemsa stained six slide specimens were prepared for each petri dish. The presence or absence of structural abnormality such as chromosome type or chromosome type gap, the presence or absence of cells (polyploid) was also observed.The test chemical did not induce chromosome aberration inChinese hamster cultured cells (CHL/IU) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In another in vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using Chinese hamster cultured cells (CHL) in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 19.1, 38.3, 76.6, 153, 306, 613, 1230, 2450µg/mL without S9 and 0, 38.3, 76.6, 153, 306, 613, 1230, 2450µg/mL with S9 in the short term treatment method and 0, 19.1, 38.3, 76.6, 153, 306, 613, 1230, 2450µg/mL in continuous treatment method. The doses for the main study were based on data from preliminary dose range finding study. Concurrent solvent and positive control plates were also included in the study. The cells were exposed to the test chemical for 24 or 48 hrs in the continuous treatment method and for 6 hrs in the short term treatment method.The test chemical did not induce chromosome aberration inChinese hamster cultured cells (CHL) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian cell gene mutation assay:
In vitro mammalian cell gene mutation assay was performed to determine the mutagenic nature of the test chemical. The study was performed using L5178Y tk+/- mouse lymphoma cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level 0, 1.6, 3.3, 6.5, 13.0 and 26.0 μg/ml in experiment 1 (with and without S9) and in experiment 2 (without S9). The doses for the main study were dependent on the dose range finding study conducted. At the end of the treatment at 13 and 26 μg/ml in all cultures of both main experiments precipitation was observed by the unaided eye. No substantial and reproducible increase of the mutant frequency was observed in both experiments. A minor increase exceeding the threshold of twice the mutant frequency of the corresponding solvent control was observed in the second culture of the second experiment. Since the historical range of negative and solvent controls was not exceeded, the effect occurred at precipitating concentrations and was not reproducible in the other culture it was considered an artificial effect due to the precipitation of the test item rather than indicating a possible mutagenic effect. Based on the observations made, the test chemical did not induce gene mutation in L5178Y tk+/- mouse lymphoma cells in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In yet another study, In vitro mammalian cell gene mutation assay was performed to determine the mutagenic nature of the test chemical. The study was performed using V79 cells with and without co-cultivation with rat hepatocytes. A test is considered to be positive if a doubling of the background mutation frequency and/or a dose dependency is observed. Based on the observations made, thetest chemical did not induce gene mutation inV79 cells and hence it is not likely to classify as a gene mutant in vitro.
Based on the data available and applying the weight of evidence approach, the test chemical does not induce gene mutation in vitro. Hence the test chemical is not likely to be mutagenic as per the criteria mentioned in CLP regulation.
Justification for classification or non-classification
Based on the data available and applying the weight of evidence approach, the test chemical does not induce gene mutation in vitro. Hence the test chemical is not likely to be mutagenic as per the criteria mentioned in CLP regulation.
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