Nickel di(acetate)

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No information on replication. Purity of test substance not reported.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity of various chemicals including nickel and cobalt compounds in cultured mouse FM3A cells.

Author:

Morita, H., M. Umeda, and H.I. Ogawa.

Year:

1991

Bibliographic source:

Mutation Research. 261:131-137.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Nakamura et al.

Principles of method if other than guideline:

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

Test material

Test material information

Constituent 1

Reference substance name:

Nickel di(acetate)

EC Number:

206-761-7

EC Name:

Nickel di(acetate)

Cas Number:

373-02-4

Molecular formula:

Ni(CH3CO2)2

IUPAC Name:

nickel di(acetate)

Specific details on test material used for the study:

- Reported as Ni(CH3COO)2-4H2O
- Source: Wako Pure Chemical Industries.

Method

Species / strain

Species / strain / cell type:

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

Controls

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 ug/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. 

Results and discussion

Test results

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).

Applicant's summary and conclusion

Executive summary:

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 ug/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. 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 4.0 x 10e-4 M.

The number of mutants per 10e6 surviving cells increased in a concentration-dependent manner, with a significant (P0.05) increase seen at 4 x 10e-4 M.

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