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EC number: 254-599-0 | CAS number: 39711-79-0
- 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 the Salmonella typhimurium strains both in the presence and absence of S9 metabolic activation system and hence is not likely to be mutagenic under the conditions of this study.
In vitro mammalian chromosome aberration study:
The test chemical did not induce chromosome aberrations in the Chinese hamster fibroblast cell line CHL and human lymphocytes in the presence and absence of S9 metabolic activation system and hence it is not mutagenic in the chromosome aberration study performed.
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 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:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- not specified
- Principles of method if other than guideline:
- WoE for the target CAS is summarized based on data from various test chemicals
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- 2. Histidine
3. Histidine - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Remarks:
- 2
- Details on mammalian cell type (if applicable):
- No data available
- 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 available
- Metabolic activation:
- with and without
- Metabolic activation system:
- (The S-9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared)
- Test concentrations with justification for top dose:
- 2. 0.00, 6.4, 32, 160, 800 µg/plate
3. 0, 6.4, 32, 160 or 800 µg/plate - Vehicle / solvent:
- 2. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Test material soluble in DMSO
3.- 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:
- other: 2-anthramine for all strains with S9 metabolic activation
- Remarks:
- 2
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- sodium azide
- other: 2-anthramine (+S9; all strains)
- Remarks:
- 3
- Details on test system and experimental conditions:
- 2. METHOD OF APPLICATION:
Standard plate
DURATION
NUMBER OF REPLICATIONS: 5 plate
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available
3.METHOD OF APPLICATION: No data
NUMBER OF REPLICATIONS: Each dosage was tested on 5 parallel plates and all the tests were performed on two separate occasions.
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data - Rationale for test conditions:
- 2. No data available
3. No data available - Evaluation criteria:
- 2. The histidine-revertant (his') colonies arising on the plates were counted
3. The plates were observed for increase in the number of revertants/plate - Statistics:
- 2. Mean number of revertants for n mumber of plates for each dose level was calculated to be the square value of mean(y) of squared roots. Standard error of mean was calculated
3.Comparisons of the number of revertants per plate were done as t-tests after a square-root transformation of each number had been performed to give homogeneity of variance. The mean number of revertants for n plates at each dose level was calculated to be the squared value of the mean (y) of the square roots. The standard error of the mean was calculated as 2 y squared root s2/n where s2 is the pooled variance of all individual square root values - Species / strain:
- S. typhimurium, other: TA98, TA100, TA1535 ,TA 1537
- Remarks:
- 2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Remarks:
- DMSO
- Untreated negative controls validity:
- valid
- Remarks:
- DMSO
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium, other: TA1537, TA1535, TA100, TA98
- Remarks:
- 3
- Metabolic activation:
- with and without
- Genotoxicity:
- not specified
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- The highest dose of 800 µg/plate showed a toxic effect on all bacterial strains
- Vehicle controls validity:
- valid
- Remarks:
- DMSO
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- 2. No data available
3. TEST-SPECIFIC CONFOUNDING FACTORS
RANGE-FINDING/SCREENING STUDIES: The toxicity of the substances was tested in the tester strains at 10-7 dilutions. Based on the results obtained, the doses were selected for the main study
ADDITIONAL INFORMATION ON CYTOTOXICITY: Test chemical did not induce gene mutation in Salmonella typhimuruim strains TA1537, TA1535, TA100, TA98 and TA97 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro. - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce mutation in the Salmonella typhimurium strains TA98, TA100, TA1535, TA 1537 and TA97 both in the presence and absence of S9 metabolic actuvation system and hence is not likely to be mutagenic under the conditions of this study.
- Executive summary:
Data available for the read across and structurally and functionally similar chemicals were reviewed to determine the mutagenic nature of the chemical. The studies are as mentioned below:
Salmonella Mutagenicity Tests of test chemical was performed in Salmonella strainTA98, TA100, TA1535, and TA 1537. Both in the presence and absence of S9 metabolic activation system.
The test chemical was dissolved in DMSO to made dose concentration of 0.00, 6.4, 32, 160, 800 µg/plate in 5 parallel plates. The S-9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared. However 2-anthramine served positive control with S9 fraction for all strain of Salmonella typhimurium and sodium azide (TA1535 and TA 100), 2-nitrofluorene (TA 1538 and TA98) served as positive control without S9 fraction.
Mutagenicity evaluated by counting histidine-revertant (his') colonies arising on these plates. As test chemical did not produce mutation in Salmonella strainTA98, TA100, TA1535, and TA 1537,Therefore it is considered to be negative for gene mutation in vitro.
In another study, Ames assay was performed to determine the mutagenic nature of test chemical. The test chemical was studies for its mutagenic nature using Salmonella typhimuruim strains TA1537, TA1535, TA100, TA98 and TA97 with and without S9 metabolic activation system. The test chemical was mixed with DMSO and used at dose level of 0, 6.4, 32, 160 or 800 µg/plate. Concurrent solvent and negative control chemicals were also included in the study. Test chemical at the highest dose of 800µg/plate showed a toxic effect on all bacterial strains. However, menthol did not induce gene mutation in Salmonella typhimuruim strains TA1537, TA1535, TA100, TA98 and TA97 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 data available for the various read across and structurally and functionally similar chemicals and applying the weight of evidence approach, the chemical did not induce mutation in the Salmonella typhimurium strain TA98, TA97, TA100, TA1535 or TA1537 both in the presence and absence of S9 metabolic activation system and hence is not likely to be mutagenic under the conditions of this study.
- 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
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Principles of method if other than guideline:
- In vitro mammalian chromosome aberration test was performed to evaluate the mutagenic nature of the test chemical
- GLP compliance:
- not specified
- Type of assay:
- other: In vitro mammalian chromosome aberration
- Target gene:
- 5. No data
6. No data - Species / strain / cell type:
- mammalian cell line, other: Chinese hamster fibroblast cell line CHL
- Remarks:
- 5
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Minimum
Essential Medium (MEM; GIBCO) supplemented by 10% calf serum
- Properly maintained: yes by 4 day passages
- Periodically checked for Mycoplasma contamination: No data available
- Periodically checked for karyotype stability: No data available
- Periodically "cleansed" against high spontaneous background: No data available - Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- lymphocytes: Human
- Remarks:
- 6
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPM1 1640 supplemented with 2 mM+glutamine (BDH), 100 U penicillin/ml, 100 pg streptomycin/ml, 10% foetal calf serum and 1% phytohaemagglutinin
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: No data
- Periodically checked for karyotype stability: No data
- Periodically "cleansed" against high spontaneous background: No data - Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- not specified
- Metabolic activation system:
- No data
- Test concentrations with justification for top dose:
- 5. At three different doses with 0.2 mg/mL being the maximum dose concentration
6. 0, 0.1, 1 or 10 mM - Vehicle / solvent:
- 5. - Vehicle(s)/solvent(s) used: Ethanol
- Justification for choice of solvent/vehicle: The chemical was soluble in ethanol
6.- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO - Untreated negative controls:
- yes
- Remarks:
- Untreated cells served as negative control
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Remarks:
- 5
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- 6
- Details on test system and experimental conditions:
- 5. METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data
- Exposure duration: 24 and 48 hrs
- Expression time (cells in growth medium): 24 and 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): Giemsa solution (1.5%, pH 6.8)
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): No data
NUMBER OF REPLICATIONS: No data
NUMBER OF CELLS EVALUATED: 100 well spread metaphases
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data
OTHER EXAMINATIONS:
- Determination of polyploidy: Yes
- Determination of endoreplication: No data
- Other: No data
OTHER: No data
6. - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO - Rationale for test conditions:
- 5. No data
6. No data - Evaluation criteria:
- 5. The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%, and positive if it was more than 10.0%.
6. The metaphase cells were observed for chromosomal aberrations - Statistics:
- 5. No data
6. Chi-square test - Species / strain:
- S. typhimurium, other: Chinese hamster fibroblast cell line CHL
- Remarks:
- 5
- Metabolic activation:
- not specified
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not specified
- Species / strain:
- lymphocytes: Human
- Remarks:
- 6
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Additional information on results:
- 50 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: The maximum dose of each sample was selected
by a preliminary test in which the dose needed
for 50% cell-growth inhibition was estimated using a cell densitometer
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data
6. 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: 10mM-menthol was chosen as the highest concentration
because higher concentrations significantly
affected the growth of human lymphocytes in
phytohaemagglutinin-stimulated cultures
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No dataTEST-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: 10mM-menthol was chosen as the highest concentration
because higher concentrations significantly
affected the growth of human lymphocytes in
phytohaemagglutinin-stimulated cultures
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- Test chemical did not induce chromosomal aberration in Chinese hamster fibroblast cell line CHL and human lymphocytes 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 from various read across and structurally and functionally similar chemicals were reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
Chromosomal aberration study was performed to determine the mutagenic nature of test chemical. The cells were exposed to the test material at three different doses with 0.2 mg/mL being the maximum concentration for 48 hr. Colcemid (final concn 0.2µg/ml) was added to the culture 2 hr before cell harvesting. The cells were then trypsinized and suspended in a hypotonic KCI solution (0.075 M) for 13 min at room temperature. After centrifugation the cells were fixed with acetic acid-methanol (1:3, v/v) and spread on clean glass slides. After air-drying, the slides were stained with Giemsa solution for 12-15 min. A hundred well-spread metaphases were observed under the microscope. In the present studies, no metabolic activation systems were applied. The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%. The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%, and positive if it was more than 10.0%. Test chemical did not induce chromosomal aberration in Chinese hamster fibroblast cell line CHL and hence 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 test chemical. The study was performed using lymphocytes isolated from the heparinized peripheral blood samples of 12 male and 12 female adult human non-smoking volunteers both with and withoutS9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose levels of 0, 0, 0.1, 1.0 or 10 mM. About 0.5-1.0 x 106isolated lymphocytes were cultured in RPM1 1640 supplemented with 2 mM+glutamine (BDH), 100 U penicillin/ml, 100 pg streptomycin/ml, 10% foetal calf serum and 1% phytohaemagglutinin. Concurrent solvent and positive control chemicals were also included in the study. All cultures were incubated in the dark at 37°C for 72 hr. Following 1 hr of exposure of the cells to colchicine the slides were prepared. Chromosomal aberrations were scored in 100 metaphase cells from each donor and tested for statistical significance by the chi-square test. The combined percentage structural aberration rate for males and females in the solvent (DMSO) control was 1.76. Lymphocyte cultures treated with 10mM-test chemical alone (10mM), had a rate of 2.11. This difference was statistically insignificant. The presence of S-9 in the culture did not influence the aberration frequency. Cultures grown in the presence of MMC showed a several-fold increase in chromosomal aberration frequency, thus validating the experimental conditions used. Based on these considerations, test chemical did not induce chromosomal aberrations in human lymphocytes 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 read acrossand structurally and functionally similar chemicals did not induce chromosomal aberration in Chinese hamster fibroblast cell line CHL and human lymphocytes 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 from various read across test chemicals was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
Ames assay:
Data available for the read across chemicals were reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
The results of the Salmonella-Escherichia coli/Mammalian-Microsome Reverse Mutation Assay with a Confirmatory Assay indicate that under the conditions of this study, the test article did not cause a positive increase in the mean number of revertants per plate with any of the tester strains in either the presence or absence of Aroclor™ induced rat liver (S9).
The test chemical was examined for its ability to cause mutagenic changes when tested in five strains of the bacteria Salmonella typhimurium, specifically, TA 1535, TA 1537, TA97, TA 98 and TA 100 through the preincubation assay method. Preliminary dose range finding study was performed initially to set the doses for the main study. The test was conducted both in the presence and absence of metabolic activation using male rat and hamster liver derived S-9 mix at dose levels of 0, 33, 100, 333, 1000, 3333 or 10000 ug/plate. The test was repeated and atleast three plates were used at each dose level. The test chemical did not induce mutation in the Salmonella typhimurium strain TA98, TA100, TA1535 or TA1537 both in the presence and absence of S9 metabolic activation system and hence is not likely to be mutagenic under the conditions of this study.
In another study, the test chemical was investigated for its ability to induce mutagenic activity when tested in an in vitro reverse mutagenicity test using four strains of the bacteria Salmonella typhimurium, specifically TA 98, TA 100, TA 1535 and TA 1537. Spot test was performed for the chemical at dose levels of 0.03, 0.3, 3 and 30 μmol/plate. The study was conducted both in the presence and absence of metabolic activation using S9 mix from Aroclor 1254 or methylcholanthrene induced rats. The test chemical is not mutagenic in the bacterium Salmonella typhimurium LT-2 strains TA 98, TA 100, TA1535 and TA37 with and without S9 metabolic activation system and hence is not likely to classify as gene mutant in vitro.
Based on the data available for the various test chemicals and applying the weight of evidence approach, the test chemical did not induce mutation in the Salmonella typhimurium strains both in the presence and absence of S9 metabolic activation system and hence is not likely to be mutagenic under the conditions of this study.
In vitro mammalian chromosome aberration study:
Data available from various test chemicals was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:
Chromosomal aberration study was performed to determine the mutagenic nature of test chemical. The cells were exposed to the test material at three different doses with 0.2 mg/mL being the maximum concentration for 48 hr. Colcemid (final concn 0.2µg/ml) was added to the culture 2 hr before cell harvesting. The cells were then trypsinized and suspended in a hypotonic KCI solution (0.075 M) for 13 min at room temperature. After centrifugation the cells were fixed with acetic acid-methanol (1:3, v/v) and spread on clean glass slides. After air-drying, the slides were stained with Giemsa solution for 12-15 min. A hundred well-spread metaphases were observed under the microscope. In the present studies, no metabolic activation systems were applied. The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%. The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%, and positive if it was more than 10.0%. Test chemical did not induce chromosomal aberration in Chinese hamster fibroblast cell line CHL and hence 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 test chemical. The study was performed using lymphocytes isolated from the heparinized peripheral blood samples of 12 male and 12 female adult human non-smoking volunteers both with and withoutS9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose levels of 0, 0, 0.1, 1.0 or 10 mM. About 0.5-1.0 x 106isolated lymphocytes were cultured in RPM1 1640 supplemented with 2 mM+glutamine (BDH), 100 U penicillin/ml, 100 pg streptomycin/ml, 10% foetal calf serum and 1% phytohaemagglutinin. Concurrent solvent and positive control chemicals were also included in the study. All cultures were incubated in the dark at 37°C for 72 hr. Following 1 hr of exposure of the cells to colchicine the slides were prepared. Chromosomal aberrations were scored in 100 metaphase cells from each donor and tested for statistical significance by the chi-square test. The combined percentage structural aberration rate for males and females in the solvent (DMSO) control was 1.76. Lymphocyte cultures treated with 10mM-test chemical alone (10mM), had a rate of 2.11. This difference was statistically insignificant. The presence of S-9 in the culture did not influence the aberration frequency. Cultures grown in the presence of MMC showed a several-fold increase in chromosomal aberration frequency, thus validating the experimental conditions used. Based on these considerations, test chemical did not induce chromosomal aberrations in human lymphocytes 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, test chemical did not induce chromosomal aberration in Chinese hamster fibroblast cell line CHL and human lymphocytes 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 data available and applying weight of evidence approach, the test chemical does not exhibit gene mutation in vitro. 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
Based on the data available and applying weight of evidence approach, the test chemical does not exhibit gene mutation in vitro. 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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