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EC number: 222-102-6 | CAS number: 3349-08-4
- 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
- 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
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Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
read across to nickel acetate (CAS 373-02-4): negative in the S. typhimurium reversion test
read across to nickel acetate (CAS 373-02-4): negative in in vitro gene mutation study in mammalian cells
read across to nickel acetate (CAS 373-02-4): negative in in vitro chromosome aberration study
read across to nickel sulphate hexahydrate (CAS 10101-97-0): positive in in vitro chromosome aberration study
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Please refer to the read across justification (IUCLID6 section 13).
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- mammalian cell line, other: Mouse mammary carcinoma cells (FM3A)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Please refer to the read-across justification (IUCLID6, section 13).
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- other: S. typhimurium strains TA98, TA100, TA1535, TA1537, and TA97, and E. Coli
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- 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
- Justification for type of information:
- Please refer to the read across justification (IUCLID6 section 13).
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- mammalian cell line, other: Syrian hamster embryo cells and human lymphocyte cultures
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not examined
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Key result
- Species / strain:
- mammalian cell line, other: C3H mouse mammary carcinoma cells (FM3A)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study without detailed documentation
- Qualifier:
- according to guideline
- Guideline:
- other: Maron and Ames 1983
- Principles of method if other than guideline:
- Study conducted according to Maron and Ames 1983. Revised methods for the Salmonella mutagenicity test. Mutation Research. 113:173-215.
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- - Name of test material: Nickel acetate
- Substance type: Pure product
- Physical state: Liquid
- Analytical purity: Reagent grade - Species / strain / cell type:
- other: S. typhimurium strains TA98, TA100, TA1535, TA1537, and TA97 and E. Coli
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9, The S9 mix was prepared from Aroclor-treated Sprague-Dawley rats, according to the procedures of Ames et al (1975).
- Test concentrations with justification for top dose:
- not reported
- Vehicle / solvent:
- - Vehicle/solvent used: dissolved or diluted in either bidistilled water or dimethyl sulfoxide (DMSO)
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: According to Ames et al. (1975)
DURATION: Details not reported, According to Ames et al. (1975)
SELECTION AGENT: Details not reported, According to Ames et al. (1975)
NUMBER OF REPLICATIONS: The assay was conducted with all strains, both in the presence and absence of S9, in duplicate or triplicate plates.
NUMBER OF CELLS EVALUATED: Details not reported, According to Ames et al. (1975)
DETERMINATION OF CYTOTOXICITY
- Method: Details not reported, According to Ames et al. (1975)
- Various dilutions of the substance were tested, starting at the solubility or cytotoxicity limit. - Evaluation criteria:
- According to Ames et al. (1975)
- Statistics:
- not reported
- Key result
- Species / strain:
- other: S. typhimurium strains TA98, TA100, TA1535, TA1537, and TA97, and E. Coli
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Additional information on results:
- In both the presence and absence of the metabolic activation system, nickel acetate was found not to be mutagenic under the conditions tested.
- Conclusions:
- The test substance nickel acetate was non-mutagenic both with and without metabolic activation in the bacterial reverse mutation assay.
- Executive summary:
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- The publication lacks information on cell and slide preparation, the purity of test material, and source of cells used. Culture conditions are not reported. Criteria for scoring chromosomal aberrations not identified. Gaps were recorded separately. Percentage of cells with chromosomal aberrations not reported. No positive control included. Low number of metaphases scored.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- FM3A cell lines used are not specified in OECD 473.
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian chromosome aberration test
- Specific details on test material used for the study:
- - Reported as Nickel acetate;
-Purchased from Wako Pure Chem. Indust. Ltd. - Species / strain / cell type:
- mammalian cell line, other: Mouse mammary carcinoma cells
- Cytokinesis block (if used):
- colcemid
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 0.20, 0.32, 0.64, and 1 mM
- Vehicle / solvent:
- Eagle's minimal essential medium with 10 % fetal bovine serum, penicillin (100U/mL) and streptomycin (100 µg/mL)
- Details on test system and experimental conditions:
- C3H mouse mammary carcinoma cells (FM3A) (1 x 10e5 cells/mL) were exposed to various concentrations of the test substance for 24 or 48 hours. Two hours prior to the end of the treatment period, colcemid was added to the culture (final concentration = 10e-7 M).
Slides were prepared using the flame-drying method. 100 metaphases were scored. Experiments were repeated. - Key result
- Species / strain:
- mammalian cell line, other: C3H mouse mammary carcinoma cells (FM3A)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Additional information on results:
- Nickel acetate induced few chromosomal aberrations and showed no dose-response.
- Conclusions:
- The test substance was negative in the chromosomal aberration study.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- In the publication there are no information about replications and the purity of the test substance given.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test using the Hprt and xprt genes)
- Qualifier:
- according to guideline
- Guideline:
- other: Nakamura et al.
- Version / remarks:
- Year: 1983 (unclear if "Year of test guideline" or "Year of study completion".)
- GLP compliance:
- not specified
- Type of assay:
- in vitro mammalian cell gene mutation test using the Hprt and xprt genes
- Specific details on test material used for the study:
- - Reported as Ni(CH3COO)2-4H2O
- Source: Wako Pure Chemical Industries - Target gene:
- HPRT, XPRT
- Species / strain / cell type:
- mammalian cell line, other: Mouse mammary carcinoma cells (FM3A)
- Metabolic activation:
- with and without
- Metabolic activation system:
- 2.5 % S15
- Test concentrations with justification for top dose:
- 0, 1.0 x 10e-4, 2.0 x 10e-4, 3.0 x 10e-4, and 4.0 x 10e-4 M
- Positive controls:
- yes
- Positive control substance:
- N-ethyl-N-nitro-N-nitrosoguanidine
- Details on test system and experimental conditions:
- FM3A cells were exposed to hypoxanthine, aminopterin, and thymidine for 1 week prior to use in the mutation assay, then cultured in Eagle's minimum essential medium supplemented with 3% fetal bovine serum, 1% non-essential amino acids, hypoxanthine, and thymidine. Cultured FM3A cells were then exposed to various concentrations of NiCl2 for 3-48 hours, washed in Hank's balanced salt solution, and then cultured for an additional 7 days. Then, 2 x 10e6 cells were inoculated into 40 mL of selective agar medium containing 10 µg/mL 6-thioguanine (6-TG) and cultured for 14 days. The number of 6TG colonies were counted and mutation frequency (per 10e6 surviving cells) was determined.
- Statistics:
- Statistical analysis was performed using the Welch test.
- Key result
- Species / strain:
- mammalian cell line, other: Mouse mammary carcinoma cells (FM3A)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Additional information on results:
- There was a concentration-dependent decrease in percent cell survival following treatment. At 0 M treatment, survival was 76%, with 100% relative plating efficiencies. This decreased to 13% survival with relative plating efficiencies of 18% after treatment at 4.0 x 10e-4 M. The number of mutants per 10e6 surviving cells increased in with Ni exposure, although results were not significant (P<0.05).
- Conclusions:
- The test substance was not mutagenic in the in vitro mammalian gene mutation test.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with national standard methods with acceptable restrictions
- Remarks:
- the publication lacks information on replication. No positive control group was included. No information is given on a vehicle (acetone) control. Only a single dose level was used for the chromosome aberration tests; no dose-response can be determined. No statistical evaluation of the data was reported. A non-standard cell line was used (embryo cells).
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Sister chromatid exchange assay and Chromosomal aberration test. No standard guideline followed.
- GLP compliance:
- not specified
- Type of assay:
- other: Sister chromatid exchange assay and Chromosomal aberration test
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Nickel Sulfate Hexahydrate, 10101-97-0
- Purity: 97 % - Species / strain / cell type:
- mammalian cell line, other: Syrian hamster embryo cells and human lymphocyte cultures
- Details on mammalian cell type (if applicable):
- Hamster embryo cells were collected after trypsinization of the embryos minus the liver and plated (density of 10e7 cells/100 mm dish) in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum.
- Metabolic activation:
- without
- Test concentrations with justification for top dose:
- 0, 1.0, 2.5, 5.0 µg/mL
- Vehicle / solvent:
- - Vehicle/solvent used: Acetone
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
Sister chromatid exchange (SCE) and chromosomal aberration tests were conducted with Syrian hamster embryo cells and human lymphocyte cultures. Hamster embryo cells were collected after trypsinization of the embryos minus the liver and plated (density of 10e7 cells/100 mm dish) in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum.
DURATION
Hamster embryo cells were incubated at 37 °C in an 11% CO2 air incubator. 24 hours later, monolayer cultures (10e6 cells/100 mm dish) were treated with nickel sulfate (prepared in acetone) and 10 µg BrdUrd/mL medium and incubated for 24 hours. Four hours prior to harvest, cells were treated with Colcemide (0.13 µg/mL), then collected, centrifuged, suspended in KCl, and fixed in methanol:glacial acetic acid.
For SCE tests, slides were prepared and stained with 4% Giemsa in Gurr's buffer solution. A minimum of 30 metaphases were scored. For chromosome aberration tests, preparations were air dried and stained with Gurr's buffer solution and 5% Giemsa. At least 125 metaphases were examined. Aberrations were scored per the International system for human cytogeneic nomenclature (1978).
For chromosome aberration tests, preparations were air dried and stained with Gurr's buffer solution and 5% Giemsa. At least 125 metaphases were examined. - Evaluation criteria:
- Aberrations were scored per the International system for human cytogeneic nomenclature (1978)
- Key result
- Species / strain:
- mammalian cell line, other: Syrian hamster embryo cells and human lymphocyte cultures
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not examined
- Untreated negative controls validity:
- valid
- Positive controls validity:
- not examined
- Additional information on results:
- NiSO4 increased the frequency of SCEs in hamster embryo cells in a dose dependent manner:
Hamster embryo cells [frequency of SCEs/metaphase (standard error)]:
BrdUrd control: 11.55 (0.84)
1.0 µg/mL: 15.95 (0.92)
2.5 µg/mL: 17.25 (1.44)
5.0 µg/mL: 21.25 (1.13)
NiSO4 also increased the number of chromosomal aberrations relative to the control:
Hamster embryo cells [percent cells with aberrations and mean aberrations/metaphase (standard error)]:
Control: 1.50%, 0.01 (0.01)
5.0 µg/mL: 16.50%, 0.16 (0.03) (Chromosomal aberrations noted in embryo cells included gaps, breaks, exchanges, and minutes) - Conclusions:
- NiSO4 was positive for chromosomal aberrations in Syrian hamster embryo cells.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (positive)
Genetic toxicity in vivo
Description of key information
read across to nickel sulphate hexahydrate (CAS 10101-97-0): positive in in vivo insect germ cell cytogenetic assay
read across to nickel sulphate hexahydrate (CAS 10101-97-0): negative in in vivo mammalian germ cell study: cytogenicity/chromosome aberration
Link to relevant study records
- Endpoint:
- in vivo mammalian germ cell study: cytogenicity / chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Please refer to the read-across justification (IUCLID6 section 13).
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Remarks:
- no cytotoxicity effects on bone marrow even at maximum tolerated dose established based on mortality and clinical observations
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Endpoint:
- genetic toxicity in vivo, other
- Remarks:
- in vivo insect germ cell cytogenetic assay
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Please refer to the read across justification (IUCLID6 section 13).
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Sex:
- male
- Genotoxicity:
- positive
- Toxicity:
- not specified
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- not examined
- Endpoint:
- in vivo mammalian germ cell study: cytogenicity / chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- GLP compliance:
- yes
- Type of assay:
- mammalian erythrocyte micronucleus test
- Specific details on test material used for the study:
- nickel sulfate hexahydrate
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Raleigh, NC, USA.
- Age at study initiation: 8 weeks
- Weight at study initiation: 239-267 g
- Fasting period before study: not reported
- Housing: 2 per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature: 64 - 79 deg F
- Humidity: humidity 30-70%
- Air changes (per hr): 10 per hr
- Photoperiod (hrs dark / hrs light): 12 hr light/dark cycle.
IN-LIFE DATES: From: April 21, 2003 To: Aug. 2003 - Route of administration:
- oral: gavage
- Vehicle:
- - Vehicle/solvent used: Cell culture control water
- Lot No: 017156 and 01100526 - Details on exposure:
- PREPARATION OF DOSING SOLUTIONS: each day the test article was prepared freshly by adding to the vehicle. Formulations were held at room temperature.
Animals were dosed by oral gavage once daily for three consecutive days, six males per dose level. The dose levels were 125, 250, or 500 mg/kg bw/day. - Duration of treatment / exposure:
- 3 days
- Frequency of treatment:
- Once daily
- Post exposure period:
- 24 hours after 3rd dose
- Dose / conc.:
- 500 mg/kg bw/day (nominal)
- Dose / conc.:
- 250 mg/kg bw/day (nominal)
- Dose / conc.:
- 125 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 6 males
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- Cyclophosphamide was used as the positive control administered by oral gavage dissolved in deionized water at a dose of 60 mg/kg bw.
- Tissues and cell types examined:
- Animals were euthanized approximately 24 hours after the third dose for extraction of bone marrow.
Blood was also collected prior to euthanization.
Nickel in bone marrow and blood was analyzed by AAS. - Details of tissue and slide preparation:
- Following centrifugation to pellet the marrow, the supernatant was spread on slides, fixed with methanol and stained in acridine orange, dried, and analyzed under fluorescent microscopy.
- Evaluation criteria:
- Slides were scored for micronuclei and to determine the PCE to NCE cell ratio. The percent micronucleated cells was determined by analyzing micronuclei from at least 2000 PCEs per animal. The criteria were those of Schmid 1976.
- Statistics:
- Data analysis was conducted using ANOVA
- Key result
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Remarks:
- no cytotoxicity effects on bone marrow even at maximum tolerated dose established based on mortality and clinical observations
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: 125-1750 mg/kg bw/day
- No cytotoxicity observed at 750 or 1000 mg/kg bw/day. Mean PCE:NCE ratios were 0.32 and 0.64 compared to 0.81 (control)
The maximum tolerated dose was estimated at 500 mg/kg bw/day
RESULTS OF DEFINITIVE STUDY
Clinical signs of toxicity were noted in all treatment animals including hypoactivity, salivation, black feces, irregular respiration, squinted/closed eyes.
No mortality occurred.
Nickel did not significantly increase micronucleated PCEs at any dose level. Nickel was not significantly cytotoxic to bone marrow at any dose level.
Dose-dependent nickel concentrations were detected in plasma and bone marrow samples.
The author's suggest the results support the non-genotoxic mode of action for soluble nickel. - Conclusions:
- The test substance nickel sulfate hexahydrate was evaluated as negative in the rat bone marrow micronucleaus assay under the conditions of this assay.
- Endpoint:
- genetic toxicity in vivo, other
- Remarks:
- in vivo insect germ cell cytogenetic assay
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- A standard Test Guideline was not specified in this study.
- GLP compliance:
- not specified
- Type of assay:
- Drosophila SLRL assay
- Specific details on test material used for the study:
- - Name of test material: Nickel sulphate hexahydrate
- Species:
- Drosophila melanogaster
- Strain:
- other: For SLRL, white males were treated and crossed to BASC females. For SCL, ring X males of the genotype XC2 y B/sc8Y were treated and crossed to y 2 w a females.
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS/ENVIRONMENTAL CONDITIONS
- Stocks maintained in mass cultures at 25 +/- 1 °C.
- Food medium was 9.2 % agar-agar, 15.4 % D-glucose, 21.5 % sugar, 38.5 % corn meal, 9.2 % yeast, 3.1 % propionic acid, 3.1 % nipagin and 1000 mL distilled water. - Route of administration:
- intraperitoneal
- Vehicle:
- - Vehicle/solvent used: 5% sucrose solution
- Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
Adult males 0-48 h old were injected i.p. with NiSO4-6H20. Treated and control males were crossed with BASC virgin females 1-3 days old for two days at a ratio of one male x two females. The progeny of each treated male was kept separately to identify possible clusters of lethals.
Brood males (A,B,C,D) were transferred to fresh virgin females at intervals of 2, 2, 3, and 3 days. F1 females fertilzied by F1 males were intercrossed in vials and the F2 scored for sex-linked recessive lethals. F3 was retested.
SCL Test: 0-2 day old ring X males, treated or untreated, were mass mated in bottles with 1-3 day old virgin females for 2 days followed by 1 or 2 successive broods, at a ratio of 3F per 1M. The F1 offspring were scored for exceptional XO males and XXY females. - Duration of treatment / exposure:
- one injection
- Frequency of treatment:
- one injection
- Post exposure period:
- 2 days
- Dose / conc.:
- 0 ppm
- Remarks:
- nominal
- Dose / conc.:
- 200 ppm
- Remarks:
- nominal
- Dose / conc.:
- 300 ppm
- Remarks:
- nominal
- Dose / conc.:
- 400 ppm
- Remarks:
- nominal
- No. of animals per sex per dose:
- 100 males treated/concentration and control
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- none reported
- Tissues and cell types examined:
- not reported
- Details of tissue and slide preparation:
- not reported
- Statistics:
- SLRL were calculated by the Kastenbaum-Bowman (1970) tables. SCL chi square was applied to test significance.
- Key result
- Sex:
- male
- Genotoxicity:
- positive
- Toxicity:
- not examined
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- not examined
- Additional information on results:
- Induction of sex-linked recessive lethals occurred at all concentrations tested. Total sex-chromosome loss was only detectable at significant numbers at the highest concentration (400 ppm). NiSO4 is mutagenic in the SLRL assay.
- Conclusions:
- NiSO4 is mutagenic in the SLRL assay and was shown to have a potential to cause total chromosome loss.
Referenceopen allclose all
Table 1: Induction of sex-linked recessive lethals in Drosophila males injected with the test substance
| A |
| B |
| C |
| D |
| A+B+C+D a |
|
Conc. [ppm] | 0-2 days | % | 2-4 days | % | 4-7 days | % | 7-10 days | % | 0-10 days | % |
Control | 4/1204 | 0.33 | 4/1436 | 0.28 | 2/926 | 0.22 | 2/1230 | 0.16 | 12/4796 | 0.25 |
200 | 8/1102 | 0.73b | 2/1024 | 0.19 | 2/1100 | 0.18 | 12/1582 | 0.76 b | 24/4818 | 0.50 b |
300 | 6/708 | 0.85b | 2/1078 | 0.19 | 2/1028 | 0.19 | 20/1376 | 1.45 b | 30/4190 | 0.72 bb |
400 | 10/1226 | 0.82b | 3/1006 | 0.30 | 2/1140 | 0.17 | 28/1700 | 1.65 b | 43/5072 | 0.85 b |
a: Sum of 2 Experiments
b: Significant at the 5% level (Kastenbaum-Bowman test).
Table 2: Frequencies of offspring resulting from sex-chromosome loss in Drosophila males injected with the test substance
|
|
| Exceptional | |||
Conc [ppm] | brood | Total offspring | Males X0 | % | Females XXY | % |
Control | A | 3560 | 3 | 0.08 | 2 | 0.06 |
| B | 3137 | 1 | 0.03 | 1 | 0.03 |
| C | 3230 | 3 | 0.09 | 1 | 0.03 |
Total | A+B+C | 9927 | 7 | 0.07 | 4 | 0.04 |
200 | A | 3269 | 8 | 0.24 | 2 | 0.06 |
| B | 3557 | 4 | 0.11 | 4 | 0.11 |
| C | 3421 | 7 | 0.20 |
|
|
Total | A+B+C | 10247 | 19 | 0.19 | 6 | 0.06 |
300 | A | 3415 | 8 | 0.23 | 2 | 0.06 |
| B | 3156 | 3 | 0.10 | 6 | 0.19 |
| C | 3260 | 8 | 0.24 | 2 | 0.06 |
Total | A+B+C | 9831 | 19 | 0.21 | 10 | 0.10 |
400 | A | 3337 | 8 | 0.24 | 2 | 0.06 |
| B | 3138 | 10 | 0.32 | 7 | 0.22 |
| C | 3212 | 8 | 0.25 | 2 | 0.11 |
Total | A+B+C | 9687 | 26 | 0.27b | 11 | 0.11 |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (positive)
Additional information
In vitro studies
In vitro gene mutation study in bacteria
No data on the substance itself is available. Therefore, read across to nickel acetate (CAS 373-02-4) was applied.
De Flora et al. (1984) evaluated the genotoxicity of multiple chemicals and chemical classes of compounds, including nickel acetate. The compounds were assayed comparatively using 1) the Ames reversion test (with his- S. typhimurium strains TAl538, TA1537, TA1535, TA98, TA100 and, in part, TA97), and a DNA-repair test. The test substance was tested with and without metabolic activation system (S9-mix). Nickel acetate was negative in the S. typhimurium reversion test for all six strains.
In vitro gene mutation study in mammalian cells
No data on the substance itself is available. Therefore, read across to nickel acetate (CAS 373-02-4) was applied (Morita et al., 1991).
FM3A cells were exposed to hypoxanthine, aminopterin, and thymidine for 1 week prior to use in the mutation assay, then cultured in Eagle's minimum essential medium supplemented with 3% fetal bovine serum, 1% non-essential amino acids, hypoxanthine, and thymidine. Cultured FM3A cells were then exposed to various concentrations of NiCl2 for 3-48 hours, washed in Hank's balanced salt solution, and then cultured for an additional 7 days. Then, 2 x 10e^6 cells were inoculated into 40 mL of selective agar medium containing 10 µg/mL 6-thioguanine (6-TG) and cultured for 14 days. The number of 6TG colonies were counted and mutation frequency (per 10e^6 surviving cells) was determined. Statistical analysis was performed using the Welch test. The positive control substance was N-methyl-N'-nitro-N-nitrosoguanidine. Cells were also tested in the presence of a metabolic activation system (2.5% S15). There was a concentration-dependent decrease in percent cell survival following treatment. At 0 M treatment, survival was 76%, with 100% relative plating efficiencies. This decreased to 7% survival with relative plating efficiencies of 9% after treatment at 4 x 10e^-4 M. The number of mutants per 10e^6 surviving cells increased in a concentration-dependent manner, with a significant increase seen at 4 x 10e^-4 M. The test substance was not mutagenic in this in vitro mammalian gene mutation test.
In vitro cytogenicity/ chromosome aberration study in mammalian cells
No data on the substance itself is available. Two studies with nickel compounds are available. Therefore, read across to nickel acetate (CAS 373-02-4) and to nickel sulphate hexahydrate (CAS 10101-97-0) was applied.
In the first study (Umeda et al., 1979), C3H mouse mammary carcinoma cells (FM3A) (1 x 10e^5 cells/mL) were exposed to various concentrations of the test substance nickel acetate for 24 or 48 hours. The study was conducted without metabolic activation. Two hours prior to the end of the treatment period, colcemid was added to the culture (final concentration = 10e^-7 M). Slides were prepared using the flame-drying method. 100 metaphases were scored. The experiments were repeated. As only few chromosomal aberrations were induced by the test substance, it was considered not to cause chromosomal aberrations.
In the second study (Larramendy et al., 1981) sister chromatid exchange (SCE) and chromosomal aberration tests were conducted with Syrian hamster embryo cells and human lymphocyte cultures. Hamster embryo cells were collected after trypsinization of the embryos minus the liver and plated (density of 10e^7 cells/100 mm dish) in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. Hamster embryo cells were incubated at 37 deg. C in an 11% CO2 air incubator. 24 hours later, monolayer cultures (10e^6 cells/100 mm dish) were treated with nickel sulphate (prepared in acetone) and 10 µg BrdUrd/mL medium and incubated for 24 hours. Four hours prior to harvest, cells were treated with Colcemide (0.13 µg/mL), then collected, centrifuged, suspended in KCl, and fixed in methanol:glacial acetic acid. For SCE tests, slides were prepared and stained with 4°% Giemsa in Gurr's buffer solution. A minimum of 30 metaphases were scored. For chromosome aberration tests, preparations were air dried and stained with Gurr's buffer solution and 5% Giemsa. At least 125 metaphases were examined. Aberrations were scored per the International system for human cytogeneic nomenclature (1978).
Human blood cultures were collected from healthy donors and diluted with Hank's balanced salt solution (without Ca and Mg). 0.5 mL of heparinized blood was mixed with 9.5 mL of RPMI 1640 medium supplemented with 20 % fetal bovine serum and 0.25 mL phytohemagglutinin M. Lymphocytes were collected by centrifugation (400 rpm for 40 minutes), resuspended in complete medium, and then washed three times with Hank's balanced salt solution. Lymphocytes were cultured in flasks at a density of 1.5 x 10e^6 cells in 5 mL RPMI 1640 medium supplemented with 20% fetal bovine serum and 0.1 mL phytohemagglutinin. 24 hours later, lymphocytes were treated with nickel sulphate (prepared in acetone) and 30 µg BrdUrd/mL medium and incubated for 48 hours. Two hours prior to harvest, cells were treated with Colcemide (0.13 µg/mL), then collected, centrifµged, suspended in KCl, and fixed in methanol:glacial acetic acid. For SCE tests, slides were prepared and stained with 4% Giemsa in Gurr's buffer solution. A minimum of 30 metaphases were scored. For chromosome aberration tests, preparations were air dried and stained with Gurr's buffer solution and 5% Giemsa. At least 125 metaphases were examined. Aberrations were scored per the International system for human cytogeneic nomenclature (1978).
NiSO4 increased the frequency of SCEs in human lymphocytes and hamster embryo cells in a dose dependent manner:
Human lymphocytes [frequency of SCEs/metaphase (standard error)]: BrdUrd control: 11.30 (0.60); 2.5 µg/mL: 17.20 (0.90); 5.0 µg/mL: 18.95 (1.52)
Hamster embryo cells [frequency of SCEs/metaphase (standard error)]: BrdUrd control: 11.55 (0.84); 1.0 µg/mL: 15.95 (0.92); 2.5 µg/mL: 17.25 (1.44); 5.0 µg/mL: 21.25 (1.13)
NiSO4 also increased the number of chromosomal aberrations relative to the control: Human lymphocytes [percent cells with aberrations and mean aberrations/metaphase (standard error)]: Control: 1.50%, 0.01 (0.01); 5.0 µg/mL: 11.20%, 0.07 (0.02), (Chromosomal aberrations noted in human lymphocytes included gaps, breaks, rings, and minutes)
Hamster embryo cells [percent cells with aberrations and mean aberrations/metaphase (standard error)]: Control: 1.50%, 0.01 (0.01); 5.0 µg/mL: 16.50%, 0.16 (0.03), (Chromosomal aberrations noted in embryo cells included gaps, breaks, exchanges, and minutes).
This summary was obtained from the ECHA disseminated dossier of nickel sulphate hexahydrate (CAS 10101-97-0).
In vivo studies
In vivo insect germ cell cytogenetic assay
No data on the substance itself is available. Therefore, read across to nickel sulphate hexahydrate (CAS 10101-97-0) was applied.
In this assay (Rodriguez-Arnaiz et Ramos, 1986), the test substance nickel sulphate hexahydrate was injected into Drosophila melanogaster males at different concentrations in order to test the chemical for the induction of SLRL (sex linked recessive lethals) and SCL (sex chromosome loss) in germ cells. The test substance was dissolved in 5% sucrose solution at 200, 300 and 400 ppm. In a preliminary test the LD50 was found to be at 400 ppm. Adult males (0-48 h old) were injected intraperitoneally with the salt. Control males were injected with 5% sucrose solution. For the SLRL test, white males (0-2 days old, treated and control) were crossed with BASC virgin females (1-3 days old), for two days at a ratio of one male × two females. The progeny of each treated male was kept separately to identify possible clusters of lethals. To test the sensitivity of different germ cell stages, a brood pattern analysis was performed (Broods A, B, C, D) by transferring the males to fresh virgin females at intervals of 2-2-3-3 days. F1 females autofertilized by F1 males were intercrossed in vials and the F2 scored for sex-linked recessive lethals. F3 was retested. For the SCL test, 0-2 day old ring X males (genotype Xc2yB/sc8Y), treated or kept untreated as controls, were mass-mated in bottles with 1-3 day old virgin y2wa females for 2 days (A) followed by 1 or 2 successive broods (B, C), at a ratio of 3 females per male. The F1 offspring were scored for exceptional XO males and XXY females. As a result, the test substance induced SLRL at all concentrations, with the peak of activity at premeiotic and postmeiotic stages. It failed to produce SCL except at the highest concentration tested, where induction of XO males was significant for the pooled data. The test substance was considered genotoxic in this assay.
In vivo mammalian germ cell study: cytogenicity/chromosome aberration
No data on the substance itself is available. Therefore, read across to nickel sulphate hexahydrate (CAS10101-97-0) was applied. The study data were obtained from the ECHA disseminated Dossier of Nickel sulphate (CAS 7786-81-4).
The study according to OECD Guideline 474 was conducted with nickel sulphate hexahydrate to evaluate its potential to induce chromosome aberrations in vivo. The study was conducted with male Sprague-Dawley rats. Each day the test article was prepared freshly by adding water. Formulations were held at room temperature. The animals were dosed by oral gavage once daily for three consecutive days. The dose levels were 125, 250, or 500 mg/kg bw/day. Cyclophosphamide was used as the positive control administered by oral gavage dissolved in deionized water at a dose of 60 mg/kg bw. The animals were euthanized approximately 24 hours after the third dose for extraction of bone marrow. Blood was also collected prior to euthanization. Nickel in bone marrow and blood was analyzed by AAS. Following centrifugation to pellet the marrow, the supernatant was spread on slides, fixed with methanol and stained in acridine orange, dried, and analyzed under fluorescent microscopy. Slides were scored for micronuclei and to determine the PCE to NCE cell ratio. The percent micronucleated cells was determined by analyzing micronuclei from at least 2000 PCEs per animal. The criteria were those of Schmid 1976. As a result, no genotoxicity was observed, and no cytotoxicity effects on bone marrow even at maximum tolerated dose established based on mortality and clinical observations.
Overall conclusion on genotoxicity
The source substance nickel sulphate hexahydrate (CAS 10101-97-0) was positive in one in vitro chromosomal aberration study. It was furthermore positive in the in vivo insect germ cell cytogenetic assay. In line with the classification of the source substances nickel acetate (CAS 373-02-4) and nickel sulphate hexahydrate (CAS 10101-97-0), the target nickel substance is classified as mutagenic into category 2. For further details, please refer to the read across justification.
Justification for classification or non-classification
Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The studies were considered reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on the results, the target and source substances are to be classified for mutagenicity into category 2 and labelled with H341 (suspected of causing genetic defects).
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