p-[4,5-dihydro-4-[[2-methoxy-5-methyl-4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]-3-methyl-5-oxo-1H-pyrazol-1-yl]benzenesulphonic acid

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 peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Primary Mutagenicity Screening Of Food Additives Currently Used In Japan

Author:

M. Ishidate, Jr, T. Sofuni, K. Yoshikawa, M. Hayashi, T. Nohmi, M. Sawada and A. Matsuoka

Year:

1984

Bibliographic source:

Fd Chem. Toxic. Vol. 22, No. 8, pp. 623-636, 1984

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

Chromosome aberration study was performed to determine the mutagenic nature of Food red 102

GLP compliance:

not specified

Type of assay:

other: In vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 1-(1-naphthylazo)-2-hydroxynaphthalene-4',6,8-trisulphonate

EC Number:

220-036-2

EC Name:

Trisodium 1-(1-naphthylazo)-2-hydroxynaphthalene-4',6,8-trisulphonate

Cas Number:

2611-82-7

Molecular formula:

C20H14N2O10S3.3Na

IUPAC Name:

trisodium 7-hydroxy-8-[(4-sulfonato-1-naphthyl)diazenyl]naphthalene-1,3-disulfonate

Details on test material:

- Name of test material: Food red 102
- Molecular formula: C20H14N2O10S3.3Na
- Molecular weight: 604.479 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Specific details on test material used for the study:

- Name of test material: Food red 102
- Molecular formula: C20H14N2O10S3.3Na
- Molecular weight: 604.479 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Method

Target gene:

No data

Species / strain

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:

not specified

Metabolic activation system:

No data

Test concentrations with justification for top dose:

At three different doses with 1 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

Controls

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: No data
- 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

Results and discussion

Test results

Species / strain:

mammalian cell line, other: Chinese hamster fibroblast cell line CHL

Metabolic activation:

not specified

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

Applicant's summary and conclusion

Conclusions:

Food red 102 did not induce chromosomal aberration in chinese hamster fibroblast cell line CHL and hence is not likely to classify as a gene mutant in vitro.

Executive summary:

Chromosomal aberration study was performed to determine the mutagenic nature of Food red 102. The cells were exposed to the test material at three different doses with 1 mg/mL being the maximum concentration for 24 and 48 hr. 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%. Food red 102 did not induce chromosomal aberration in chinese hamster fibroblast cell line CHL and hence is not likely to classify as a gene mutant in vitro.