4-[(5-amino-3-methyl-1-phenyl-1H-pyrazol-4-yl)azo]-2,5-dichlorobenzenesulphonic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

supporting study

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:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Principles of method if other than guideline:

In vitro mammalian chromosome aberration test was performed to evaluate the mutagenic response of tartrazine

GLP compliance:

not specified

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

EC Number:

217-699-5

EC Name:

Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Cas Number:

1934-21-0

Molecular formula:

C16-H12-N4-O9-S2.3Na

IUPAC Name:

trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate

Constituent 2

Reference substance name:

trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo) pyrazole-3-carboxylate

IUPAC Name:

trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo) pyrazole-3-carboxylate

Details on test material:

- Name of test material (as cited in study report): Tartrazine
- Molecular formula (if other than submission substance): C16H12N4O9S2.3Na.
C16-H9-N4-O9-S2.3Na
- Molecular weight (if other than submission substance): 534.3681 g/mol
- Substance type: Organic
- Physical state: No data available
Purity No data available
- Impurities (identity and concentrations): No data available

Specific details on test material used for the study:

- Name of test material: Tartrazine
- IUPAC name: trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo) pyrazole-3-carboxylate
- Molecular formula: C16H12N4O9S2.3Na.
C16H9N4O9S2.3Na
- Molecular weight: 534.3681 g/mol
- Substance type: Organic
- Physical state: No data available
- Purity No data available
- Impurities: No data available

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:

without

Metabolic activation system:

Metabolic activation system

Test concentrations with justification for top dose:

At three different doses with 2500 µg/plate being the maximum dose concentration

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Physiological saline
- Justification for choice of solvent/vehicle: The test chemical was soluble in physiological saline

Controls

Untreated negative controls:

yes

Remarks:

Untreated cells

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 available
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): 48 hrs
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hr

SELECTION AGENT (mutation assays): No data available

SPINDLE INHIBITOR (cytogenetic assays): No data available

STAIN (for cytogenetic assays): 1.5% Geimsa stain at pH 6.8

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: 100 well spread metaphases were observed

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: Yes

- Determination of endoreplication: No data available

- Other: The incidence 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

OTHER: No data available

Rationale for test conditions:

No data

Evaluation criteria:

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 available

Results and discussion

Test results

Species / strain:

other: Chinese hamster fibroblast cell line

Metabolic activation:

without

Genotoxicity:

positive

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:

No data

Remarks on result:

other:

Applicant's summary and conclusion

Conclusions:

The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical tartrazine was considered to be more than 10% in the absence of metabolic activation system during the 48 hrs study duration and hence tartrazine is mutagenic in vitro.

Executive summary:

Chromosomal aberration test was performed for the test chemical tartrazine using Chinese hamster fibroblast cell line CHL. The cells were exposed to the test material at three different doses with 2.5 mg/plate being the highest dose for 24 and 48 hr. 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%. The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical tartrazine was considered to be more than 10% in the absence of metabolic activation system during the 48 hrs study duration and hence tartrazine is mutagenic in vitro.