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Toxicological information

Genetic toxicity: in vitro

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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:
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

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
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:
other: Chinese hamster fibroblast cell line (CHL)
Details on mammalian cell lines (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 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:
- Vehicle(s)/solvent(s) used: Physiological saline
- Justification for choice of solvent/vehicle: The test chemical was soluble in physiological saline
Controls
Negative controls:
yes
Remarks:
Untreated cells
Solvent controls:
yes
Remarks:
Physiological saline
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Details on test system and 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:
not specified
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
not specified
Remarks on result:
other:
Additional information on results:
No data

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.