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EC number: 247-019-2 | CAS number: 25481-21-4
- 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
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- Endpoint summary
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- 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
Gene mutation in vitro:
Data available for the read across chemicals was reviewed to determine the mutagenic nature of the target chemical nickel(2+) ion 2-({2-[(carboxylatomethyl)(carboxymethyl)amino]ethyl}(carboxymethyl)amino)acetate (CAS no 25481 -21 -4). The studies are as mentoned below:
Ames test:
The test chemical did not induce gene mutation in the Salmonella typhimurium TA98, TA100, TA1535, TA1537, and TA1538 both in the presence and absence of S9 activation system and hence the chemical is not likely to be a gene mutant.
In vitro mammalian 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 gene mutation study in mammalian cells:
Test chemical did not induce mutation in mammalian cell line in the presence and absence of metabolic activation and hence it is not likely to classify as a gene mutant in vitro.
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Remarks:
- Read across data
- 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 473 (In Vitro Mammalian Chromosome Aberration Test)
- Principles of method if other than guideline:
- Chromosomal aberration study was performed to determine the mutagenic nature of test chemical.
- GLP compliance:
- not specified
- Type of assay:
- other: Chromosome aberration assay
- 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:
- No data
- Test concentrations with justification for top dose:
- At three different doses with 0.25 mg/mL being the maximum dose concentration
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Physiological saline
- Justification for choice of solvent/vehicle: The chemical was soluble in Physiological saline - Untreated negative controls:
- yes
- Remarks:
- Untreated cells served as negative control
- 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
- 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 - Rationale for test conditions:
- No data
- Evaluation criteria:
- 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%.
- Statistics:
- No data
- Species / strain:
- S. typhimurium, other: Chinese hamster fibroblast cell line CHL
- Metabolic activation:
- without
- Genotoxicity:
- negative
- 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:
- 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 - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- Test chemical did not induce chromosomal aberration in Chinese hamster fibroblast cell line CHL and hence it is not likely to classify as a gene mutant in vitro.
- Executive summary:
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 0.25 mg/mL being the maximum concentration for 24hr and 48 hrs. 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 it is not likely to classify as a gene mutant in vitro.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Remarks:
- Read across data
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- “The study contains experimental data of a read-across analogue
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The study was conducted to investigate genetic toxicity of food flavouring ingredients.
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian cell gene mutation tests using the thymidine kinase gene
- Target gene:
- Thymidine Kinase
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- The cell line was maintained in Fischer's medium containing 10% horse serum, antibiotics,
glutamine, sodium pyruvate, and Pluronic F68 . - Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat liver S9 activation system.
- Test concentrations with justification for top dose:
- 125 µl/ml
- Untreated negative controls:
- yes
- True negative controls:
- yes
- Positive controls:
- not specified
- Details on test system and experimental conditions:
- The procedures described by Clive et al . (1979) . The TK+/- -3 .7 .2C heterozygote of the L5178Y mouse lymphoma cell line was maintained in Fischer's medium containing 10% horse serum, antibiotics, glutamine, sodium pyruvate, and Pluronic F68 . In a typical assay procedure, the thymidine kinase competent heterozygote was exposed to the test article in both the presence and absence of an
induced rat liver S9 and cofactors (CORE) . After a 4 hour exposure period, the cells were washed and
incubated at 37° C for 48 hours to allow phenotypic expression before cloning 3x10 cells in Noble agar
containing the selective agent trifluorothymidine or bromodeoxyuridine . - Evaluation criteria:
- Colonies were counted after 10-14 days' growth using an automatic colony counter. Mutant frequency was determined by calculating the ratio of mutant to viable colonies cloned without selective medium .
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- When L5178Y mouse lymphoma cells were treated with myristic acid, it has not induced any genetic mutation in presence or absence of S9 activation medium.
- Executive summary:
The study was conducted to access the genetic mutation property of myristic acid. Mouse lymphoma cell line assay was performed in presence and absence of Aroclor 1254-induced rat liver S9 activation system.
The cell line was maintained in Fischer's medium containing 10% horse serum, antibiotics, glutamine, sodium pyruvate, and Pluronic F68. In the assay procedure, the thymidine kinase competent heterozygote was exposed to the test article in both the presence and absence of an induced rat liver S9 and cofactors (CORE) . After a 4 hour exposure period, the cells were washed and incubated at 37° C for 48 hours to allow phenotypic expression before cloning 3x10 cells in Noble agar containing the selective agent trifluorothymidine or bromodeoxyuridine . Colonies were counted after 10-14 days' growth using an automatic colony counter. Mutant frequency was determined by calculating the ratio of mutant to viable colonies cloned without selective medium .
Results indicated that, when L5178Y mouse lymphoma cells were treated with myristic acid, it has not induced any genetic mutation in presence or absence of S9 activation medium.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- 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:
- The Ames salmonella typhimurium mutagenicity test was conducted for the test chemical
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- Histidine
- Species / strain / cell type:
- S. typhimurium, other: TA98, TA100, TA1535, TA1537, and TA1538
- 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:
- Liver S9 homogenate was prepared from male Sprague-Dawley rats and Syrian golden hamsters that had been injected with Aroclor 1254 at 500 mg/kg body weight
- Test concentrations with justification for top dose:
- 33-10000 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Yes, no detailed data available
- Justification for choice of solvent/vehicle: No data available - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- no detailed data available
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: No data available
- Exposure duration: 48 h
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available
SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available
NUMBER OF REPLICATIONS: Triplicate
NUMBER OF CELLS EVALUATED: No data available
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available
OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available
OTHER: No data available - Rationale for test conditions:
- No data
- Evaluation criteria:
- The criteria used to evaluate a test were as follows: for a test article to be considered positive, it had to induce at least a doubling (TA98, TA100, and TA1535) in the mean number of revertants per plate of at least one tester strain. This increase in the mean revertants per plate had to be accompanied by a dose response to increasing concentrations of the test chemical. If the study showed a dose response with a less than 3-fold increase on TA1537 or TA1538, the response had to be confirmed in a repeat experiment.
- Statistics:
- No data available
- Species / strain:
- S. typhimurium, other: TA98, TA100, TA1535, TA1537, and TA1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- 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 doses that were tested in the mutagenicity assay were selected based on the levels of cytotoxicity observed in a preliminary dose range-finding study using strain TA100. Ten dose levels of the chemical, one plate per dose, were tested in both the presence and the absence of induced hamster S9. If no toxicity was observed, a total maximum dose of 10 mg of test chemical per plate was used.
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce gene mutation in the Salmonella typhimurium TA98, TA100, TA1535, TA1537, and TA1538 both in the presence and absence of S9 activation system and hence the chemical is not likely to be a gene mutant.
- Executive summary:
Ames mutagenicity test was conducted for the test chemical to evaluate its genetoxic effects when exposed to Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538 with dose concentration of 33-10000 µg/plate in plate incorporation assay. Based on the preliminary study conducted, the test compound was used at a five dose level from 33-10000 µg/plate. The plates were incubated for 48 h at 37±2 °C. Five doses of test chemical, together with the appropriate concurrent solvent and positive controls, were tested in triplicate on each tester strain without metabolic activation and also with activation by induced rat and hamster liver S9 preparations. For a test article to be considered positive, it had to induce at least a doubling (TA98, TA100, and TA1535) in the mean number of revertants per plate of at least one tester strain. This increase in the mean revertants per plate had to be accompanied by a dose response to increasing concentrations of the test chemical.
The test chemical did not induce gene mutation in the Salmonella typhimurium TA98, TA100, TA1535, TA1537, and TA1538 both in the presence and absence of S9 activation system and hence the chemical is not likely to be a gene mutant.
Referenceopen allclose all
Table: Mutagenicity of the test chemical in chromosomal aberration test in vitro
Max dose |
Solvent |
Polyploid (%) |
Structural aberrations |
Result |
|
% |
hr |
||||
0.25 |
Physiological saline |
1.0 |
3.0 |
24 |
- |
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:
Ames test:
Data available for the read across chemicals was reviewed to determine the mutagenic nature of the target chemical nickel(2+) ion 2-({2-[(carboxylato methyl) (carboxymethyl)amino]ethyl}(carboxymethyl)amino)acetate (CAS no 25481 -21 -4). The studies are as mentoned below:
Study 1 :
Ames mutagenicity test was conducted for the test chemical to evaluate its genetoxic effects when exposed to Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538 with dose concentration of 33-10000 µg/plate in plate incorporation assay. Based on the preliminary study conducted, the test compound was used at a five dose level from 33-10000 µg/plate. The plates were incubated for 48 h at 37±2 °C. Five doses of test chemical, together with the appropriate concurrent solvent and positive controls, were tested in triplicate on each tester strain without metabolic activation and also with activation by induced rat and hamster liver S9 preparations. For a test article to be considered positive, it had to induce at least a doubling (TA98, TA100, and TA1535) in the mean number of revertants per plate of at least one tester strain. This increase in the mean revertants per plate had to be accompanied by a dose response to increasing concentrations of the test chemical. The test chemical did not induce gene mutation in the Salmonella typhimurium TA98, TA100, TA1535, TA1537, and TA1538 both in the presence and absence of S9 activation system and hence the chemical is not likely to be a gene mutant.
Study 2 :
Ames assay was performed to determine the mutagenic nature of the test chemical EthylenediamineTetraacetic acid. The study was performed using Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli strain WP2uvrA in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 1.22- 5000 µg/plate by the preincubation method.The test chemical EthylenediamineTetraacetic acid did not induce gene mutation in the Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and E. coli strain WP2uvrA in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
In vitro mammalian chromosome aberration study:
1. 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 0.25 mg/mL being the maximum concentration for 24hr and 48 hrs. 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 it is not likely to classify as a gene mutant in vitro.
2. In vitro mammalian chromosome aberration test was performed for the given test chemical using Chinese hamster fibroblast cell line CHL. The cells were exposed to the test material at 1.0 mg/ml for 48 hr. 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 given test chemical does not induce chromosomal aberrations in the Chinese hamster fibroblast cell line and hence is negative for gene mutation in vitro.
In vitro gene mutation study in mammalian cells:
The study was conducted to access the genetic mutation property of myristic acid. Mouse lymphoma cell line assay was performed in presence and absence of Aroclor 1254-induced rat liver S9 activation system.
The cell line was maintained in Fischer's medium containing 10% horse serum, antibiotics, glutamine, sodium pyruvate, and Pluronic F68. In the assay procedure, the thymidine kinase competent heterozygote was exposed to the test article in both the presence and absence of an induced rat liver S9 and cofactors (CORE) . After a 4 hour exposure period, the cells were washed and incubated at 37° C for 48 hours to allow phenotypic expression before cloning 3x10 cells in Noble agar containing the selective agent trifluorothymidine or bromodeoxyuridine . Colonies were counted after 10-14 days' growth using an automatic colony counter. Mutant frequency was determined by calculating the ratio of mutant to viable colonies cloned without selective medium .
Results indicated that, when L5178Y mouse lymphoma cells were treated with myristic acid, it has not induced any genetic mutation in presence or absence of S9 activation medium.
Justification for classification or non-classification
Based on the data available for the read across chemicals, the target chemical nickel(2+) ion 2-({2-[(carboxylatomethyl) (carboxymethyl)amino] ethyl} (carboxymethyl)amino)acetate (CAS no 25481 -21 -4) 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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