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EC number: 272-939-6 | CAS number: 68921-42-6
- 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 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.
Chromosome aberration study:
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 mutagenic in the chromosome aberration study performed.
in vitro mammalian gene mutation:
The given test material does not induce mutation in the L5178Y (Tk+/-) mouse lymphoma cells in the absence of S9 metabolic activation system.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Remarks:
- Read across data
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Data available from study report.
- Qualifier:
- according to guideline
- Guideline:
- other: OECD guideline No. 476 and the ICH guidelines S2A and S2B
- Principles of method if other than guideline:
- The mutagenic potential of Patent Blue V was assessed in the in vitro mammalian cell gene mutation assay using L5178Y (Tk+/-) mouse lymphoma cells.
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- In vitro mammalian cell gene mutation assay using L5178Y (Tk+/-) mouse lymphoma cells
- Species / strain / cell type:
- other: L5178Y (Tk+/-) mouse lymphoma cells
- Details on mammalian cell type (if applicable):
- No data available
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Test concentrations with justification for top dose:
- 0.86 to 2500 μg/ml
- Vehicle / solvent:
- No data available
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- not specified
- Details on test system and experimental conditions:
- Details on test system and conditions
METHOD OF APPLICATION: In medium
- Exposure duration: 24 hrs
NUMBER OF REPLICATIONS: No detailed data available
DURATION
- Exposure duration: 24 hrs
Number of Replications: Duplicate - Evaluation criteria:
- No data
- Statistics:
- No data
- Species / strain:
- other: L5178Y (Tk+/-) mouse lymphoma cells
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no 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: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative without metabolic activation
The given test material does not induce mutation in the L5178Y (Tk+/-) mouse lymphoma cells in the absence of S9 metabolic activation system. - Executive summary:
The mutagenic potential of Patent Blue V was assessed in the in vitro mammalian cell gene mutation assay using L5178Y (Tk+/-) mouse lymphoma cells. The study was conducted in compliance with the OECD guideline No. 476 and the ICH guidelines S2A and S2B. Long treatment (approximately 24 hours) without metabolic activation was performed. Replicate cultures were set up at each experimental point. No statistically or biologically significant increases in the mutant frequency were noted up to the maximum tested dose level of 2500 μg/ml at any treatment time in the absence of metabolic activation. Patent Blue V does not induce mutation in the L5178Y (Tk+/-) mouse lymphoma cellsin the absence of S9 metabolic activation system. Hence the test material is not likely to classify as a gene mutant in vitro.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer reviewed publication
- Qualifier:
- equivalent or similar 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 response of the test chemical
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- No data
- Species / strain / cell type:
- mammalian cell line, other: Chinese hamster fibroblast cell line (CHL)
- 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
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- without
- Metabolic activation system:
- Metabolic activation system
- Test concentrations with justification for top dose:
- At three different doses with 2500 µg/plate being the maximum dose concentration
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Physiological saline
- Justification for choice of solvent/vehicle: The test chemical was soluble in physiological saline - Untreated negative controls:
- yes
- Remarks:
- Untreated cells
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Physiological saline
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: No data available
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hr
SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): 1.5% Geimsa stain at pH 6.8
NUMBER OF REPLICATIONS: No data available
NUMBER OF CELLS EVALUATED: 100 well spread metaphases were observed
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available
OTHER EXAMINATIONS:
- Determination of polyploidy: Yes
- Determination of endoreplication: No data available
- Other: The incidence 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
OTHER: No data available - Rationale for test conditions:
- No data
- Evaluation criteria:
- 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%
- Statistics:
- No data available
- Species / strain:
- other: Chinese hamster fibroblast cell line
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not specified
- Additional information on results:
- No data
- Conclusions:
- The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical was considered to be more than 10% in the absence of metabolic activation system during the 48 hrs study duration.
- Executive summary:
Chromosomal aberration test was performed for the test chemical using Chinese hamster fibroblast cell line CHL. The cells were exposed to the test material at three different doses with 2.5 mg/plate being the highest dose for 24 and 48 hr. 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%. The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical was considered to be more than 10% in the absence of metabolic activation system during the 48 hrs study duration and was mutagenic in the absence of S9 metabolic activation system.
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- 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
- Justification for type of information:
- Data is from peer reviewed publication
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Principles of method if other than guideline:
- Gene mutation toxicity study was performed to determine the mutagenic nature of test chemical.
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Histidine
- Species / strain / cell type:
- other: TA 92. TA 1535, TA 100, TA 1537, TA 94, TA 98
- 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
- Metabolic activation system:
- The liver microsome fraction (S-9) was prepared from the liver of Fischer rats
- Test concentrations with justification for top dose:
- 6 different concentrations were used; 5.0 mg/plate was the maximum concentration
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Phosphate buffer
- Justification for choice of solvent/vehicle: The test chemical was soluble in Phosphate buffer - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Phosphate buffer
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 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: 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 result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated).
- Statistics:
- No data
- Species / strain:
- S. typhimurium, other: TA 92. TA 1535, TA 100, TA 1537, TA 94, TA 98
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Additional information on results:
- The maximum dose for negative results represents the highest non-cytotoxic dose used in the experiment
- Remarks on result:
- other: No mutagenic potential
- Conclusions:
- Test chemical did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
- Executive summary:
Gene mutation toxicity study was performed to determine the mutagenic nature of test chemical. The study was performed using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without S9 metabolic activation system. The test was performed as per the preincubation assay at six different concentrations with 5.0 mg/plate being the maximum concentration. The chemical was dissolved in phosphate buffer. Preincubation was performed for 20 mins and the exposure duration was for 48 hrs. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). The test chemical did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 in the presence and absence of S9 metabolic activation system and hence the chemical 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
Ames Test:
Data available for the test chemicals were reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:
Gene mutation toxicity study was performed to determine the mutagenic nature of test chemical. The study was performed using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without S9 metabolic activation system. The test was performed as per the preincubation assay at six different concentrations with 5.0 mg/plate being the maximum concentration. The chemical was dissolved in phosphate buffer. Preincubation was performed for 20 mins and the exposure duration was for 48 hrs. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). The test chemical did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 in the presence and absence of S9 metabolic activation system and hence the chemical 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 test chemical. The study was performed as per the qualitative plate test using Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA98 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in glass distilled water or DMSO and used at dose levels of 0, 50, 250 or 1000 µg/plate. Concurrent solvent, negative, historical and positive control chemicals were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA98 in the presence and absence of S9 metabolic activation system and hence it is considered to be non-mutagenic.
Gene mutation toxicity study was performed to determine the mutagenic nature of test chemical. The study was performed as per the qualitative plate technique using Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA98 in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in glass distilled water or DMSO and used at dose levels of 0, 50, 250 or 1000 µg/plate. Concurrent solvent, negative, historical and positive control chemicals were also included in the study. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA98 in the presence and absence of S9 metabolic activation system and hence it is considered to be non mutagenic.
Based on the data available, the test chemical did not induce gene mutation in sallmonela strain in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Chromosomal Abberation:
Data available for the test chemicals were reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:
Chromosomal aberration test was performed for the test chemical using Chinese hamster fibroblast cell line CHL. The cells were exposed to the test material at three different doses with 2.5 mg/plate being the highest dose for 24 and 48 hr. 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%. The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical was considered to be more than 10% in the absence of metabolic activation system during the 48 hrs study duration and was mutagenic in the absence of S9 metabolic activation system.
In another study, Chromosomal aberration tests were carried out to determine the mutagenic nature of the test chemical. The study was performed using Chinese hamster ovary cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in medium and used at dose levels of up to 5 mg/mL. In the assays for chromosomal aberrations, the top dose (TD) was based on toxicity, solubility, or the upper testing limit (5 mg/ml). In the chromosome aberration assay without activation, cells were exposed to the test chemical for 8 hr. The test chemical was washed off, and the cells were treated with 0.1 µg/ml Colcemid for 2-2.5 hr. With metabolic activation, the cells were exposed to the test chemical plus the metabolic activation mixture for 2 hr, washed, incubated for 8 hr, and then treated with Colcemid for 2-2.5 hr. A delayed harvest was used in the aberration assay. In these tests the cell growth period was extended to about 20 hr. Air-dried slides were coded and stained with Giemsa. One hundred to 200 cells from each of the three highest scorable doses were analyzed. All aberrations were individually classified (e.g., chromatid breaks, chromosome breaks, triradials, etc.). These data were combined as the percent of cells with simple (deletions), complex (exchanges), and total (simple, complex and other) aberrations. The test chemical did not induce chromosome aberrations in Chinese hamster ovary cells (CHO) 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 test was also performed to determine the mutagenic nature of test chemical. The study was performed using Chinese hamster ovary cells in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in serum free culture medium and used at dose level of 0, 1000, 1600, 3000 or 5000 µg/plate. Concurrent solvent and negative control chemicals were also included in the study. Approximately 24 hr prior to cell treatment, 1.2 x 106cells were seeded per 75 cm2 flask. For assays without metabolic activation, the cells were treated for about 10 hr. Colcemid was added 2-3 hr prior to cell harvest by mitotic shake-off. In the test protocol for assays with metabolic activation cells were harvested approximately 11 hr after removal of the S9 fraction. Colcemid was added 2 hr prior to harvest. Slides were stained in 6% Giemsa for 5-10 min. One hundred cells were scored for each dose in early studies and 200 cells per dose in later studies. All slides except high-dose positive controls were coded. Only metaphase cells in which the chromosome number was between 19 and 23 were scored. The test chemical did not induce ABS at doses up to 5000µg/ml in either the presence or the absence of S9. Based on the observations made, the test chemical did not induce chromosome aberration in Chinese hamster ovary cells in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
in vitro mammalian gene mutation:
The mutagenic potential of Patent Blue V was assessed in the in vitro mammalian cell gene mutation assay using L5178Y (Tk+/-) mouse lymphoma cells. The study was conducted in compliance with the OECD guideline No. 476 and the ICH guidelines S2A and S2B. Long treatment (approximately 24 hours) without metabolic activation was performed. Replicate cultures were set up at each experimental point. No statistically or biologically significant increases in the mutant frequency were noted up to the maximum tested dose level of 2500 μg/ml at any treatment time in the absence of metabolic activation. Patent Blue V does not induce mutation in the L5178Y (Tk+/-) mouse lymphoma cellsin the absence of S9 metabolic activation system. Hence the test material is not likely to classify as a gene mutant in vitro.
Based on the observations made, the test chemical did not induce mammalian gene mutation in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
Justification for classification or non-classification
Based on the data available and applying weight of evidence approach, the given test chemical does not exhibit gene mutation in vitro by Ames assay, In vitro mammalian chromosome aberration study and in vitro mammalian gene mutation assay. 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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