[carbonato(2-)]tetrahydroxytrinickel

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 2008 to October 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

[carbonato(2-)]tetrahydroxytrinickel

EC Number:

235-715-9

EC Name:

[carbonato(2-)]tetrahydroxytrinickel

Cas Number:

12607-70-4

Molecular formula:

Ni3(OH)4CO3

IUPAC Name:

trinickel monocarbonate tetrahydroxide

Constituent 2

Reference substance name:

pentanickel dicarbonate hexahydroxide

Cas Number:

12122-15-5

Molecular formula:

C2H6Ni5O12

IUPAC Name:

pentanickel dicarbonate hexahydroxide

Details on test material:

- Name of test material (as cited in study report): Nickel hydroxycarbonate
- Substance type: pure product
- Physical state: green powder
- Analytical purity: 100%
- Stability under test conditions: Stable
- Storage condition of test material: room temperature, closed container

Method

Target gene:

Thymidine Kinase (TK) locus

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: RPMI + 3% horse serum, or + 7.5% horse serum for long-term exposure
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: not reported
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

Experiment I: with metabolic activation: 10, 40, 80, 110, 140, 170, 200, 230 ug/mL
Experiment I: without metabolic activation: 10, 20, 40, 60, 80, 110, 140, 170 ug/mL
Experiment II: with metabolic activation: 40, 80, 100, 120, 140, 160, 200, 220 ug/mL
Experiment II: with metabolic activation: 1, 2, 5, 10, 17.5, 25, 32.5, 40 ug/ml

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: [medium]

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: EMS or MMS

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 1-3 days
- Exposure duration: 4 or 24 hours
- Expression time (cells in growth medium): 48 or 72 hours

NUMBER OF CELLS EVALUATED: 200 cells per well in four 96 wells

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

Positive results based on clear dose related mutant frequency and biologically relevant response.

Statistics:

no statistics performed

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

ADDITIONAL INFORMATION ON CYTOTOXICITY:
-Growth inhibition was observed with and without metabolic activation.
-pH was within the physiological range

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Nickel hydroxycarbonate is considered to be non-mutagenic in the mouse lymphoma thymidine kinase locus assay.

Executive summary:

BSL (2008) performed a study using the mouse lymphoma cell line L5178Y to assess the potential for nickel hydroxycarbonate to cause gene mutations in vitro. Genetic damage was assessed with the thymidine kinase assay (carried out according to OECD Guideline 476 and using GLP standards). Two separate experiments were conducted, both with and without metabolic activation using a S9 mix. Based on results obtained from a pre-experiment, the exposure concentrations were as follows: Experiment I with metabolic activation (10, 40, 80, 110, 140, 170, 200, and 230 μg/mL), without metabolic activation (10, 20, 40, 60, 80, 110, 140,and 170 μg/mL); Experiment II with metabolic activation (40, 80, 100, 120, 140, 160, 200, and 220 μg/mL), without metabolic activation (1, 2, 5, 10, 17.5, 25, 32.5, and 40 μg/mL). In Experiment II without metabolic activation, cells were exposed for 24 hours as a long-term exposure assay; the other experiments used a 4-hour exposure period. Negative controls (vehicle only) and positive controls(ethylmethanesulfonate, methylmethanesulfonate [both without metabolic activation]; benzo[a]pyrene [with metabolic activation]) were used in the study.

For each experiment, mutagenicity, toxicity (measured as growth inhibition), and colony size (to determine clastogenic effects) data were collected. No biologically significant increase in mutation frequencies occurred following nickel hydroxycarbonate treatment, for either experiment and under conditions with or without metabolic activation. Growth inhibition occurred in Experiments I and II, under conditions with and without metabolic activation. The relative total growth (RTG) findings with the corresponding highest concentrations evaluated were as follows: Experiment I with metabolic activation (13.49% for 230 μg/mL), Experiment I without metabolic activation (13.43% for 170 μg/mL), Experiment II with metabolic activation (17.12% for 220 ug/mL), Experiment II without metabolic activation (12.67%. for 40 ug/mL). No clastogenic effects occurred following nickel hydroxycarbonate treatment, for either

experiment and under conditions with or without metabolic activation. The authors concluded that, using the cell line L5178Y, nickel hydroxycarbonate was non-mutagenic at the mouse lymphoma thymidine kinase locus.

STUDY RATED BY AN INDEPENDENT REVIEWER