Iron(II) fumarate

Genetic toxicity: in vitro

Administrative data

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:

data from handbook or collection of data

Justification for type of information:

Data is from publication.

Data source

Reference

Reference Type:

publication

Title:

Primary mutagenicity screening of the given test chemical

Author:

Ishidate et al.

Year:

1984

Bibliographic source:

Food Chem Toxicol

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

in vitro mammalian chromosome aberration study was conducted for the given test chemical.

GLP compliance:

not specified

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Fumaric acid

EC Number:

203-743-0

EC Name:

Fumaric acid

Cas Number:

110-17-8

Molecular formula:

C4H4O4

IUPAC Name:

(2E)-but-2-enedioic acid

Test material form:

solid: crystalline

Details on test material:

Name of the test chemical: Fumaric acid
IUPAC name: (2E)-but-2-enedioic acid
Molecular Formula: C4H4O4
Molecular Weight: 116.072 g/mol
Substance Type: Organic
Physical State: Solid

Method

Target gene:

No data

Species / strain

Species / strain / cell type:

other: Chinese hamster lung-derived fibroblasts (CHL)

Details on mammalian cell type (if applicable):

CELLS USED
- Type and source of cells: The cell line was originally established from the lung of a newborn female at the Cancer Research Institute, Tokyo (Koyama, Utakoji & Ono, 1970),

For cell lines:
- Absence of Mycoplasma contamination: No data
- Number of passages if applicable: 4-day passages
- Methods for maintenance in cell culture: The cell line was maintained in Minimum Essential Medium (MEM; GIBCO) supplemented by 10% calf serum.
- Cell cycle length, doubling time or proliferation index : The doubling time was approximately 15 hr.
- Modal number of chromosomes: The modal chromosome number is 25

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

Colcemid (final conc 0.2 microgm/ml) was added to the culture 2 hr before cell harvesting.

Metabolic activation:

without

Metabolic activation system:

no metabolic activation systems were applied

Test concentrations with justification for top dose:

0, 0.5 µg/ml. Top dose was expected to produce a 50% inhibition of cell growth based on data from a pre-experiment.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used [none; no data; acetone; arachis oil; beeswax; carbowaxe; castor oil; cetosteryl alcohol; cetyl alcohol; CMC (carboxymethyl cellulose); coconut oil; corn oil; cotton seed oil; DMSO; ethanol; glycerol ester; glycolester; hydrogenated vegetable oil; lecithin; macrogel ester; maize oil; olive oil; paraffin oil; peanut oil; petrolatum; physiol. saline; poloxamer; polyethylene glycol; propylene glycol; silicone oil; sorbitan derivative; soya oil; theobroma oil; vegetable oil; aqueous solvents (water or saline or culture medium)] : Physiological saline

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

DURATION
- Exposure duration: 24 & 48 hours
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
NUMBER OF CELLS EVALUATED: 100
DETERMINATION OF CYTOTOXICITY
- Method: 50% cell growth inhibition
OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

Results were considered negative if incidence of aberrations was less than 4.9%, equivocal if it was between 5 and 9.9% and positive if it was more than 10%

Statistics:

No data

Results and discussion

Test results

Species / strain:

other: Chinese hamster lung-derived fibroblasts (CHL)

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

Chromosome aberration test (CA) in mammalian cells:

o Changes in ploidy (polyploidy cells and cells with endoreduplicated chromosomes) if seen: 0% polypliody was observed.

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Results of the chromosomal aberration study:

Polypliody(%)	Structural aberrations (%)	hours
0	1	24

Applicant's summary and conclusion

Conclusions:

The test chemical tested negative for mutachromosomal effects in Chinese hamster lung-derived fibroblasts (CHL).

Executive summary:

In vitro mammalian chromosome aberration study was conducted for the given test chemical in Chinese hamster lung-derived fibroblasts (CHL) in the absence of metabolic activation system. The cell line was originally established from the lung of a newborn female at the Cancer Research Institute, Tokyo (Koyama, Utakoji & Ono, 1970), and was maintained by 4-day passages in Minimum Essential Medium (MEM; GIBCO) supplemented by 10% calf serum. The modal chromosome number is 25 and the doubling time was approximately 15 hr. The cells were exposed to chemical at three concentrations up to 0.5 µg/ml for 24 and 48 hr. Physiological saline was used as solvent. The top dose was expected to produce a 50% inhibition on cell growth based on data from a pre-experiment. Colcemid (final conc 0.2 µg/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 (1.5%, at pH 6.8; E. Merck) for 12-15 min. A hundred well-spread metaphases were observed under the microscope (x 600 with a no cover objective lens). 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. Results were considered negative if incidence of aberrations was less than 4.9%, equivocal if it was between 5 and 9.9% and positive if it was more than 10%. The test chemical tested negative for mutachromosomal effects in Chinese hamster lung-derived fibroblasts (CHL).