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EC number: 232-234-6 | CAS number: 7790-94-5
- 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
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Ames test:
The test chemical did not induce gene mutation in Salmonella typhimurium and E.coli 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 chromosome aberration study:
The test chemical induced chromosomal aberration in the Chinese hamster ovary K1 (CHO-K1) cell line used in the presence and absence of S9 metabolic activation system and hence it is likely to be clastogenic in vitro.
In vitro mammalian cell gene mutation assay:
The test chemical did not induce mammalian cell gene mutation assay in Fischer L5178Y 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.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- 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 various test chemicals
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- equivalent or similar 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:
- 1. Histidine
2. Streptomycin
3. Histidine - Species / strain / cell type:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
- Remarks:
- 1
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- E. coli, other: B/Sd-4/1,3,4,5 and B/Sd-4/3,4
- Remarks:
- 2
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Remarks:
- 3
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- Rat liver S9 metabolic activation system
- Test concentrations with justification for top dose:
- 1. 0, 0.001, 0.01, 0.1, 1.0, 5.0 uL/plate
2. 0, 0.002, 0.002, 0.002, 0.003 or 0.005 %
3. trial 1 and 2: 10 - 1000 ug/plate
trial 2a: 10 - 5000 ug/plate
trial 2b: 100 - 3333 ug/plate
(only with TA 98 / TA 100 without S9-mix) - Vehicle / solvent:
- 1. - Vehicle(s)/solvent(s) used: Deionized water
- Justification for choice of solvent/vehicle: The test chemical was soluble in Deionized water
2. - Vehicle(s)/solvent(s) used: Distilled water
- Justification for choice of solvent/vehicle: The test chemical was soluble in Distilled water
3. - Vehicle(s)/solvent(s) used: ethylene glycol dimethylether (EGDE)
- Justification for choice of solvent/vehicle: Test chemical solubility in solvent - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Deionized water
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- other: NF (TA98, TA1538; -S9), QM (TA1537; -S9), 2 AA (All strains; +S9)
- Remarks:
- 1
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Distilled water
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Remarks:
- 2
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethylene glycol dimethylether (EGDE)
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- other: 4-nitro-o-phenylene-diamine (TA 98, TA 1537; -S9) and 2-aminoanthracene (TA 98, TA 100, TA 1535, TA 1537; +S9)
- Remarks:
- 3
- Details on test system and experimental conditions:
- 1. METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: No data
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): 48 hrs
- 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: One
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: 3 hrs
- Expression time:
Streptomycin agar plates: 48 hrs (2 days)
Sterptomycin free plates: 6 days
- 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: No data
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
3. METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: No data
- Exposure duration: No data
- Expression time: No data
- 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: Triplicate
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 - Rationale for test conditions:
- No data
- Evaluation criteria:
- 1. The result was considered positive if the test material produced a positive dose-related response over three concentrations with the lowest increase equal to three times the spontaneous revertant rate for TA1535, TA1537 or TA1538 strains while for TA98 and TA100 strains a doubling of the rate at the highest increase over three dose levels was considered positive.
2. The streptomycin agar and free plates were scored for the presence of revertant colonies and the frequency of mutants was calculated by dividing the number of colonies by the number of viable bacteria plated
3. The plates were observed for number of revertants/plate - Statistics:
- 1. No data
2. Poisson distribution was used to estimate the variance and the statistical significance of the result was evaluated
3. No data - Species / strain:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
- Remarks:
- 1
- 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
- Species / strain:
- E. coli, other: B/Sd-4/1,3,4,5 and B/Sd-4/3,4
- Remarks:
- 2
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium, other: TA 98, TA 1535, TA 1537
- Remarks:
- 3
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ≥3333 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- 3
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ≥3333 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- 3
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- ≥3333 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- 1. No data
2. TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no data on pH values given
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: 5000 ug/plate
- Other confounding effects: No data
RANGE-FINDING/SCREENING STUDIES: No data
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data
3. No data - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce gene mutation in Salmonella typhimurium and E.coli 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:
Bacterial gene mutation assay was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in deionized water and used at dose level of 0, 0.001, 0.01, 0.1, 1.0, 5.0 uL/plate. The study was performed as per the plate incorporation assay and the number of revertant colonies was scored after incubation at 37°C for 48 hr. The result was considered positive if the test material produced a positive dose-related response over three concentrations with the lowest increase equal to three times the spontaneous revertant rate for TA1535, TA1537 or TA1538 strains while for TA98 and TA100 strains a doubling of the rate at the highest increase over three dose levels was considered positive.The test chemical did not induce gene mutation in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538 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 study, gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using E. coli Strains B/Sd-4/1,3,4,5 and B/Sd-4/3,4. The test chemical was dissolved in distilled water and used at dose levels of 0, 0.002, 0.002, 0.002, 0.003 or 0.005 %. For every experiment, bacteria were grown for 24 hours at 37˚C in an aerated broth culture containing 10 micrograms of streptomycin per milliliter. They reached a saturation titer of approximately 2 to 3 x 109cells per milliliter. Each culture was started from an inoculum, usually large, taken from streptomycin-agar slants kept in a refrigerator. Before treatment the bacteria were washed in saline and resuspended in distilled water. A sample of the new suspension was added to the desired solution of chemical in distilled water, and incubated at 37˚C for a certain period of time; no growth occurs under these conditions. Another sample of the same suspension was added to an equal amount of distilled water and incubated for the same period of time, as a control. At the end of the treatment period, both treated and control suspensions were assayed by plating suitable dilutions on streptomycin-agar plates. At the same time they were plated (0.1 ml per Petri dish), either undiluted or diluted not more than 1:10 in plain broth, onto a number of streptomycin-free plates, using a glass spreader and a turntable. The assay plates were incubated for 48 hours, after which it was possible to count the colonies and calculate the titers of the two suspensions at the end of treatment, and the percentage of survivors. The streptomycin-free plates (mutant plates) were incubated for at least six days. After this time the colonies were scored, and the frequency of mutants calculated by dividing the number of colonies by the number of (viable) bacteria plated. The test chemical did not increase the frequency of revertant mutants in the strains of E. coli used. The test chemical did not induce gene mutation in E. coli Strains B/Sd-4/1,3,4,5 and B/Sd-4/3,4 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 TA 98, TA 100, TA 1535, TA 1537 in the presence and absence of S9 metabolic activation system. The test chemical was soluble inethylene glycol dimethylether (EGDE) and used at dose level of 10 - 1000 ug/plate in trial 1 and 2, 10 - 5000 ug/plate in trial 2a and 100 - 3333 ug/plate in trial 2b with TA 98 / TA 100 without S9-mix. Concurrent solvent and positive control chemicals were also included in the study. Significant and reproducible dose-dependent increase up to the highest investigated concentration only in TA 100 without S9-mix. No effects were observed in the strains TA 98, TA 1535 and TA 1537. Based on the observations made, the test chemical did not induce gene mutation inSalmonella typhimurium TA 98, TA 1535, TA 1537 in the presence and absence of S9 metabolic activation system. It however induced gene mutation in the strain TA100 in the absence of S9 metabolic activation system. Overall the test chemical is not likely to classify as a gene mutant in vitro.
Based on the data available, the test chemical did not induce gene mutation in Salmonella typhimurium and E.coli 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:
- read-across from supporting substance (structural analogue or surrogate)
- 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 various test chemicals
- Reason / purpose for cross-reference:
- read-across source
- 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:
- No data
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Remarks:
- 1
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Cells were grown in Fischer's Medium for mouse leukaemic cells supplemented with 10% horse serum, sodium pyruvate and penicillin-streptomycin. The cloning medium consisted of growth medium with 20% horse serum and 0.37% agar.
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: Yes, Cell stocks were determined to be mycoplasma free.
- 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:
- Chinese hamster Ovary (CHO)
- Remarks:
- CHO-K1 / 2 / 3
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Ham's F12 medium supplemented with sodium bicarbonate (16.7 mM), kanamycin (60 µg/ml) and 10% fetal calf serum (FCS).
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: Yes, The cells were confirmed to be free of mycoplasma.
- 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:
- Liver S-9 homogenate prepared from random bred male CD-1 mice
- Test concentrations with justification for top dose:
- 1. Without activation : 0, 0.1, 0.2, 0.4 uL/mL (1.2, 2.4, 4.8 mM)
With activation : 0, 0.2, 0.4, 0.8 uL/mL (2.4, 4.8, 9.6 mM)
2. With S9: 0, 2, 4 , 6 or 8 mM
Without S9: 0, 2, 4 , 6 , 8 or 10 mM
3. With S9: 0, 6, 8, 10 or 12 mM
Without S9: 0, 8, 10, 12, 14 or 16 mM - Vehicle / solvent:
- 1. - Vehicle(s)/solvent(s) used: Ethanol
- Justification for choice of solvent/vehicle: The test chemical was soluble in ethanol
2./3.
- Vehicle(s)/solvent(s) used: No data
- Justification for choice of solvent/vehicle: No data - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Ethanol
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- N-dimethylnitrosamine
- ethylmethanesulphonate
- 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./3.
- Details on test system and experimental conditions:
- 1. METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data
- Exposure duration: 4 hrs
- Expression time (cells in growth medium): 20 hrs
- Selection time (if incubation with a selection agent): 3 hrs
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): Colchicine
STAIN (for cytogenetic assays): Giemsa stain
NUMBER OF REPLICATIONS: One
NUMBER OF CELLS EVALUATED: 50 cells at each dose level
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./3. METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data
- Exposure duration: With S9: 6 hrs
Without S9: 24 hrs
- Expression time: 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): No data
STAIN (for cytogenetic assays): No data
NUMBER OF REPLICATIONS: No data
NUMBER OF CELLS EVALUATED: 100 metaphases were evaluated
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:
- 1. Compounds that induced aberrations in 8% or more of the cells were considered to possess clastogenic (chromosome damaging) potential.
2./3. The cell line was observed for chromosome aberration studies - Statistics:
- 1. In chromosome aberration studies, a value of 8% or more of the cells with aberrations was considered the lower limit of significance based on a concurrent and historical control frequency.
2./3. 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:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- Chinese hamster Ovary (CHO)
- Remarks:
- CHO-K1 / 2 / 3
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- 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 chromosome aberrations in the 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 of the test chemical. The studies are as mentioned below:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using Fischer L5178Y mouse-lymphoma cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in ethanol and used at dose level of 0, 0.1-0.4 and 0.2-0.8µg/mL. The cell line was exposed to the test chemical for 4 hrs. BUdR was added to treated cells for 20 hr, or two cell doublings, to permit its incorporation into the DNA. The cells were then prepared for Chromosome aberration staining. Metaphase arrest was accomplished by adding colchicine (0.5µg/ml final concentration) to the growing cultures during the last 3 hr of incubation. The cells were harvested with 0.075 M-hypotonic KCl solution followed by fixing in three changes of Carnoy's fixative (methanol-glacial acetic acid, 3:1). The rinsed slides were then air-dried overnight. 50 metaphase cells were evaluated for chromosome aberration. Concurrent solvent and positive control chemicals were also included in the study. Based on the observations made,The test chemical did not induce chromosome aberrations inFischer L5178Y 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 another study, in vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of two test chemicals. The study was performed using Chinese hamster ovary K1 (CHO-K1) cells in the presence and absence of S9 metabolic activation system. The cell line was maintained in Ham's F12 medium supplemented with sodium bicarbonate (16.7 mM), kanamycin (60µg/ml) and 10% fetal calf serum (FCS). The cultures were incubated at 37°C in an atmosphere of 5°70 CO2 in air and subcultures were made every 3-4 days. The cells were confirmed to be free of mycoplasma. One test chemical was tested at dose level of 0, 2, 4, 8 or 10 mM in the absence of S9 and 0, 2, 4, 6 or 8 mM in the presence of S9. Another test chemical was tested at concentration of 0, 6, 8, 10 or 12 mM in the presence of S9 metabolic activation system and at dose level of 0, 8, 10, 12, 14 or 16 mM.The test chemical was added to the F12 medium, and subsequently the pH of the medium was measured. The pH of the medium was measured prior to and after treatment using a pH meter. In the absence of S9 mix,the test chemical was added to the 3-day-old cultures after a medium change for 24 hrs. In metabolic activation with S9 mix, cells were treated with S9 mix and the test chemical for 6 h. Recovery times were 18 h. The cells were then cultured for 24 h. In all experiments, chromosome preparations were made 24 h after the start of treatment. The number of cells with chromosomal aberrations was recorded on 100 metaphases. Chromatid breaks and exchanges were observed. Based on the observations made, the test chemical induced chromosomal aberration in the Chinese hamster ovary K1 (CHO-K1) cell line used in the presence and absence of S9 metabolic activation system and hence it is likely to be clastogenic in vitro. The results of the study indicate that low pH can cause chromosomal effects in this assay; findings seen with the inorganic acids tested are therefore considered to be artefactual.
Based on the observations made, the test chemical induced chromosomal aberration in the Chinese hamster ovary K1 (CHO-K1) cell line used in the presence and absence of S9 metabolic activation system and hence it is likely to be clastogenic in vitro.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- 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 variuos test chemicals
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- equivalent or similar 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 test
- Target gene:
- Tk locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Remarks:
- Fischer cells / 1
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Cells were grown in Fischer's Medium for mouse leukaemic cells supplemented with 10% horse serum, sodium pyruvate and penicillin-streptomycin. The cloning medium consisted of growth medium with 20% horse serum and 0.37% agar.
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: Yes, Cell stocks were determined to be mycoplasma free.
- 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:
- mouse lymphoma L5178Y cells
- Remarks:
- 2
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Stock cultures were maintained in Fischer's medium for leukemic cells of mice at approx. 37 °C on gyratory tables. Fischer's medium (F0) was supplemented with 2 mM glutamine (HEM), 110 µg/ml sodium pyruvate 0.05% Pluronic F68 and and 10% by volume of heat-inactivated donor horse 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
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- Liver S-9 homogenate prepared from random bred male CD-1 mice
- Test concentrations with justification for top dose:
- 1. Without activation: 0.4, 0.5, 0.6, 0.7, 0.8 µL/mL (4.8-9.6 mM)
With activation: 0.8, 1.0, 1.2, 1.4, 1.6 µL/mL (9.6-23.0 mM)
2. With metabolic activation: pH 6.3, 6.6, 6.9, 7.2, 7.5
Without metabolic activation: pH 6.0, 6.2, 6.5, 6.7, 6.9, 7.0 - Vehicle / solvent:
- 1. - Vehicle(s)/solvent(s) used: Water
- Justification for choice of solvent/vehicle: The test chemical was soluble in water
2. No data - Untreated negative controls:
- yes
- Remarks:
- Medium
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Water
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- other: With activation: Dimethylnitrosoamine 0.3 µL/mL
- 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
- Details on test system and experimental conditions:
- 1. METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data
- Exposure duration: 4 hrs
- Expression time (cells in growth medium): 3 days
- Selection time (if incubation with a selection agent): 10 days
- 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: One
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 medium
DURATION
- Preincubation period: No data
- Exposure duration: 4 hrs
- Expression time: 2 days
- 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: 100 metaphases were evaluated
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:
- 1. A dose-response relationship over three concentrations of the four dose levels employed is observed, or an increase in the mutation frequencies at the highest dose is at least 2.5 times greater than the solvent control value.
2. The cell line was observed for gene mutation at tk locus - Statistics:
- No data
- Species / strain:
- mouse lymphoma L5178Y cells
- Remarks:
- Fischer cells / 1
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- With metabolic activation: 0.8 µL/mL (100% growth inhibition) and Without metabolic activation: Ca 1.0 µL/mL (12.5% growth inhibition at 0.8 µL/mL)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- mouse lymphoma L5178Y cells
- Remarks:
- 2
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- 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 mammalian cell gene mutation assay in Fischer L5178Y 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.
- 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:
In vitro mammalian cell gene mutation assay was performed to determine the mutagenic nature of the test chemical. The study was performed using Fischer L5178Y mouse-lymphoma cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in water and used at dose level of 0.4, 0.5, 0.6, 0.7, 0.8 uL/mL (4.8-9.6 mM) in the presence of S9 and 0.8, 1.0, 1.2, 1.4, 1.6 uL/mL (9.6-23.0 mM) in the absence of S9. The cell line was exposed to the test chemical for 4 hrs in the presence and absence of S9 metabolic activation system. The culture was allowed to express the TK- mutation in growth medium for 3 days prior to mutant determination. Cells were counted daily and diluted to a concentration of 3 x 105cells/ml, considered optimal for growth. After 3 days, an aliquot of 3 x 106cells at each dose was seeded into the selection medium and cultures were incubated for 10 days. Mutant colonies (resistant to BUdR) were stained and counted in an Artek electronic colony counter, set to discriminate colonies 200 pm or larger in size. A portion of this first aliquot culture was diluted to give 300 cells/plate prior to the addition of BUdR and seeded into non-selective medium to measure cell survival. Mutant cell counts from the selection plates were corrected for cell survival and expressed as a mutant frequency/clonable cell. Based on the observations made, thetest chemical did not induce mammalian cell gene mutation assay in Fischer L5178Y 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 mouse lymphoma L5178Y cells in the presence and absence of S9 metabolic activation system. The test chemical was used at pH 6.3, 6.6, 6.9, 7.2, 7.5 in the presence of S9 metabolic activation system and at pH 6.0, 6.2, 6.5, 6.7, 6.9, 7.0 in the absence of S9 metabolic activation system.A series of sterile, 15-ml conical centrifuge tubes were inoculated with 3 x 106 cells per tube. The cells were sedimented by centrifugation at approx. 150 x g for 10 min. The cells were then resuspended in medium that had been adjusted to the appropriate pH. RPMI 1640 medium containing 10% heat-inactivated horse serum was used for assays in which the pH was controlled with organic buffers. All other experiments were suspended in F10 medium. The total volume for treatment was 10 ml; for activation assays, the 10-ml volume included the activation mix.The dosed tubes were placed in a shaker incubator at approximately 80 rpm at 37 ° C. After 4 h, the cells were washed twice and resuspended in 10 ml of F10 and returned to the shaker incubator. A standard expression time of 2 days was used. On day 1, the viable cell count was determined for each culture and the cell densities were adjusted to 3 × 105cells/ml. On day 2, each tube was adjusted to contain a 10-ml cell suspension at 3 × 106cells/ml. A sample containing 3 × 106 cells was added to 100 ml of Fl0 medium containing trifluorothymidine (TFT) and approx. 0.3% Noble agar (Difco, Detroit, MI). Each sample was then poured into a set of three 100-nm culture dishes (Falcon). A sample of 300 cells from each culture was also seeded into a set of three 100-mm culture dishes in F10 medium containing 0.3% agar without any selective agent in order to determine cloning efficiencies. The dishes were then incubated for 10-12 days in a 37°C incubator with 5% CO2/humidified atmosphere for colony development. At the end of the incubation period, colonies were scored on an Artek 880 electronic colony counter. Based on the observations made,the test chemical induced gene mutation at tk locus inmouse lymphoma L5178Y cells in the presence and absence of S9 metabolic activation system and hence it is likely to be mutagenic in vitro. The results of the study indicate that low pH can cause gene mutation in this assay; findings seen with the inorganic acids tested are therefore considered to be artefactual.
Based on the observations made, the test chemical did not induce mammalian cell gene mutation assay in Fischer L5178Y 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.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Gene mutation in vitro:
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 test:
Bacterial gene mutation assay was performed to determine the mutagenic nature of the test chemical 1. The study was performed using Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in deionized water and used at dose level of 0, 0.001, 0.01, 0.1, 1.0, 5.0 uL/plate. The study was performed as per the plate incorporation assay and the number of revertant colonies was scored after incubation at 37°C for 48 hr. The result was considered positive if the test material produced a positive dose-related response over three concentrations with the lowest increase equal to three times the spontaneous revertant rate for TA1535, TA1537 or TA1538 strains while for TA98 and TA100 strains a doubling of the rate at the highest increase over three dose levels was considered positive.The test chemical did not induce gene mutation in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538 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 study, gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical 2. The study was performed using E. coli Strains B/Sd-4/1,3,4,5 and B/Sd-4/3,4. The test chemical was dissolved in distilled water and used at dose levels of 0, 0.002, 0.002, 0.002, 0.003 or 0.005 %. For every experiment, bacteria were grown for 24 hours at 37˚C in an aerated broth culture containing 10 micrograms of streptomycin per milliliter. They reached a saturation titer of approximately 2 to 3 x 109cells per milliliter. Each culture was started from an inoculum, usually large, taken from streptomycin-agar slants kept in a refrigerator. Before treatment the bacteria were washed in saline and resuspended in distilled water. A sample of the new suspension was added to the desired solution of chemical in distilled water, and incubated at 37˚C for a certain period of time; no growth occurs under these conditions. Another sample of the same suspension was added to an equal amount of distilled water and incubated for the same period of time, as a control. At the end of the treatment period, both treated and control suspensions were assayed by plating suitable dilutions on streptomycin-agar plates. At the same time they were plated (0.1 ml per Petri dish), either undiluted or diluted not more than 1:10 in plain broth, onto a number of streptomycin-free plates, using a glass spreader and a turntable. The assay plates were incubated for 48 hours, after which it was possible to count the colonies and calculate the titers of the two suspensions at the end of treatment, and the percentage of survivors. The streptomycin-free plates (mutant plates) were incubated for at least six days. After this time the colonies were scored, and the frequency of mutants calculated by dividing the number of colonies by the number of (viable) bacteria plated. The test chemical did not increase the frequency of revertant mutants in the strains of E. coli used. The test chemical did not induce gene mutation in E. coli Strains B/Sd-4/1,3,4,5 and B/Sd-4/3,4 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 3. The study was performed using Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 in the presence and absence of S9 metabolic activation system. The test chemical was soluble inethylene glycol dimethylether (EGDE) and used at dose level of 10 - 1000 ug/plate in trial 1 and 2, 10 - 5000 ug/plate in trial 2a and 100 - 3333 ug/plate in trial 2b with TA 98 / TA 100 without S9-mix. Concurrent solvent and positive control chemicals were also included in the study. Significant and reproducible dose-dependent increase up to the highest investigated concentration only in TA 100 without S9-mix. No effects were observed in the strains TA 98, TA 1535 and TA 1537. Based on the observations made, the test chemical did not induce gene mutation inSalmonella typhimurium TA 98, TA 1535, TA 1537 in the presence and absence of S9 metabolic activation system. It however induced gene mutation in the strain TA100 in the absence of S9 metabolic activation system. Overall the test chemical is not likely to classify as a gene mutant in vitro.
Based on the data available, the test chemical did not induce gene mutation in Salmonella typhimurium and E.coli 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 chromosome aberration study:
Data available for the various test chemicals was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical 1. The study was performed using Fischer L5178Y mouse-lymphoma cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in ethanol and used at dose level of 0, 0.1-0.4 and 0.2-0.8µg/mL. The cell line was exposed to the test chemical for 4 hrs. BUdR was added to treated cells for 20 hr, or two cell doublings, to permit its incorporation into the DNA. The cells were then prepared for Chromosome aberration staining. Metaphase arrest was accomplished by adding colchicine (0.5µg/ml final concentration) to the growing cultures during the last 3 hr of incubation. The cells were harvested with 0.075 M-hypotonic KCl solution followed by fixing in three changes of Carnoy's fixative (methanol-glacial acetic acid, 3:1). The rinsed slides were then air-dried overnight. 50 metaphase cells were evaluated for chromosome aberration. Concurrent solvent and positive control chemicals were also included in the study. Based on the observations made,The test chemical did not induce chromosome aberrations inFischer L5178Y 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 another study, in vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of two test chemicals 1 and 2. The study was performed using Chinese hamster ovary K1 (CHO-K1) cells in the presence and absence of S9 metabolic activation system. The cell line was maintained in Ham's F12 medium supplemented with sodium bicarbonate (16.7 mM), kanamycin (60µg/ml) and 10% fetal calf serum (FCS). The cultures were incubated at 37°C in an atmosphere of 5°70 CO2 in air and subcultures were made every 3-4 days. The cells were confirmed to be free of mycoplasma. One test chemical was tested at dose level of 0, 2, 4, 8 or 10 mM in the absence of S9 and 0, 2, 4, 6 or 8 mM in the presence of S9. Another test chemical was tested at concentration of 0, 6, 8, 10 or 12 mM in the presence of S9 metabolic activation system and at dose level of 0, 8, 10, 12, 14 or 16 mM.The test chemical was added to the F12 medium, and subsequently the pH of the medium was measured. The pH of the medium was measured prior to and after treatment using a pH meter. In the absence of S9 mix,the test chemical was added to the 3-day-old cultures after a medium change for 24 hrs. In metabolic activation with S9 mix, cells were treated with S9 mix and the test chemical for 6 h. Recovery times were 18 h. The cells were then cultured for 24 h. In all experiments, chromosome preparations were made 24 h after the start of treatment. The number of cells with chromosomal aberrations was recorded on 100 metaphases. Chromatid breaks and exchanges were observed. Based on the observations made, the test chemical induced chromosomal aberration in the Chinese hamster ovary K1 (CHO-K1) cell line used in the presence and absence of S9 metabolic activation system and hence it is likely to be clastogenic in vitro. The results of the study indicate that low pH can cause chromosomal effects in this assay; findings seen with the inorganic acids tested are therefore considered to be artefactual.
Based on the observations made, the test chemical induced chromosomal aberration in the Chinese hamster ovary K1 (CHO-K1) cell line used in the presence and absence of S9 metabolic activation system and hence it is likely to be clastogenic in vitro.
In vitro mammalian cell gene mutation study:
In vitro mammalian cell gene mutation assay was performed to determine the mutagenic nature of the test chemical 1. The study was performed using Fischer L5178Y mouse-lymphoma cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in water and used at dose level of 0.4, 0.5, 0.6, 0.7, 0.8 uL/mL (4.8-9.6 mM) in the presence of S9 and 0.8, 1.0, 1.2, 1.4, 1.6 uL/mL (9.6-23.0 mM) in the absence of S9. The cell line was exposed to the test chemical for 4 hrs in the presence and absence of S9 metabolic activation system. The culture was allowed to express the TK- mutation in growth medium for 3 days prior to mutant determination. Cells were counted daily and diluted to a concentration of 3 x 105cells/ml, considered optimal for growth. After 3 days, an aliquot of 3 x 106cells at each dose was seeded into the selection medium and cultures were incubated for 10 days. Mutant colonies (resistant to BUdR) were stained and counted in an Artek electronic colony counter, set to discriminate colonies 200 pm or larger in size. A portion of this first aliquot culture was diluted to give 300 cells/plate prior to the addition of BUdR and seeded into non-selective medium to measure cell survival. Mutant cell counts from the selection plates were corrected for cell survival and expressed as a mutant frequency/clonable cell. Based on the observations made, thetest chemical did not induce mammalian cell gene mutation assay in Fischer L5178Y 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 2. The study was performed using mouse lymphoma L5178Y cells in the presence and absence of S9 metabolic activation system. The test chemical was used at pH 6.3, 6.6, 6.9, 7.2, 7.5 in the presence of S9 metabolic activation system and at pH 6.0, 6.2, 6.5, 6.7, 6.9, 7.0 in the absence of S9 metabolic activation system.A series of sterile, 15-ml conical centrifuge tubes were inoculated with 3 x 106cells per tube. The cells were sedimented by centrifugation at approx. 150 x g for 10 min. The cells were then resuspended in medium that had been adjusted to the appropriate pH. RPMI 1640 medium containing 10% heat-inactivated horse serum was used for assays in which the pH was controlled with organic buffers. All other experiments were suspended in F10 medium. The total volume for treatment was 10 ml; for activation assays, the 10-ml volume included the activation mix.The dosed tubes were placed in a shaker incubator at approximately 80 rpm at 37 ° C. After 4 h, the cells were washed twice and resuspended in 10 ml of F10 and returned to the shaker incubator. A standard expression time of 2 days was used. On day 1, the viable cell count was determined for each culture and the cell densities were adjusted to 3 × 105cells/ml. On day 2, each tube was adjusted to contain a 10-ml cell suspension at 3 × 106cells/ml. A sample containing 3 × 106 cells was added to 100 ml of Fl0 medium containing trifluorothymidine (TFT) and approx. 0.3% Noble agar (Difco, Detroit, MI). Each sample was then poured into a set of three 100-nm culture dishes (Falcon). A sample of 300 cells from each culture was also seeded into a set of three 100-mm culture dishes in F10 medium containing 0.3% agar without any selective agent in order to determine cloning efficiencies. The dishes were then incubated for 10-12 days in a 37°C incubator with 5% CO2/humidified atmosphere for colony development. At the end of the incubation period, colonies were scored on an Artek 880 electronic colony counter. Based on the observations made,the test chemical induced gene mutation at tk locus inmouse lymphoma L5178Y cells in the presence and absence of S9 metabolic activation system and hence it is likely to be mutagenic in vitro. The results of the study indicate that low pH can cause gene mutation in this assay; findings seen with the inorganic acids tested are therefore considered to be artefactual.
Based on the observations made, the test chemical did not induce mammalian cell gene mutation assay in Fischer L5178Y 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.
On the basis of observations made and applying the weight of evidence approach, the test chemical does not exhibit gene mutation in vitro. This chemical undergoes immediate hydrolysis in water of body fluids after contact with any tissue. The hydrolysis products are test chemical 1 and 2. Some mutagenic effects though observed but results of the studies performed with inorganic acids test chemical 1 and 2 indicate that low pH can cause false positive effects in these assay. Hence further testing is not scientifically justified. Hence the test chemical is not likely to classify as a gene mutant as per the criteria mentioned in CLP regulation.
Justification for classification or non-classification
On the basis of observations made and applying the weight of evidence approach, the test chemical does not exhibit gene mutation in vitro. This chemical undergoes immediate hydrolysis in water of body fluids after contact with any tissue. The hydrolysis products are test chemical 1 and 2. Some mutagenic effects though observed but results of the studies performed with inorganic acids test chemical 1 and 2 indicate that low pH can cause false positive effects in these assay. Hence further testing is not scientifically justified. Hence the test chemical is not likely to classify as a gene mutant as per the criteria mentioned in CLP regulation.
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