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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
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:
Methodology for the testing of food dyes for genotoxic activity: experiments with RED 2G (C.I. 18050)
Author:
R.B. Haveland-Smith, R.D. Combes, B.A. Bridges
Year:
1979
Bibliographic source:
Mutation Research, 64 , 241—248

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: Refer below principle
Principles of method if other than guideline:
Evaluation of mutagenicity of Red 2G in Salmonella typhimurium strain TA1538 in a Fluctuation test.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Constituent 1
Reference substance name:
Red 2G
IUPAC Name:
Red 2G
Constituent 2
Chemical structure
Reference substance name:
Disodium 5-acetylamino-4-hydroxy-3-(phenylazo)naphthalene-2,7-disulphonate
EC Number:
223-098-9
EC Name:
Disodium 5-acetylamino-4-hydroxy-3-(phenylazo)naphthalene-2,7-disulphonate
Cas Number:
3734-67-6
Molecular formula:
C18H13N3Na2O8S2
IUPAC Name:
disodium 5-acetamido-4-hydroxy-3-(phenyldiazenyl)naphthalene-2,7-disulfonate
Test material form:
not specified
Details on test material:
- Name of test material: Red 2G
- Molecular formula: C18H15N3O8S2.2Na
- Molecular weight: 509.426 g/mol
- Substance type: Organic
- Physical state: Aqueous solutions or suspensions
- Purity: 80%
- Impurities (identity and concentrations): As, Pb ,Sb, BaSO4, Cr, Cu, Zn,Free aromatic amines, Synthetic intermediates (excluding free aromatic amines),Subsidiary colours were present.
Specific details on test material used for the study:
- Name of test material: Red 2G
- Molecular formula: C18H15N3O8S2.2Na
- Molecular weight: 509.426 g/mol
- Substance type: Organic
- Physical state: Aqueous solutions or suspensions
- Purity: 80%
- Impurities (identity and concentrations): As, Pb ,Sb, BaSO4, Cr, Cu, Zn,Free aromatic amines, Synthetic intermediates (excluding free aromatic amines),Subsidiary colours were present.

Method

Target gene:
Histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1538
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
other: his rfa uvrB
Species / strain / cell type:
E. coli WP2 uvr A
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S9 mix consisted of calcium-precipitated microsome from liver of male Sprague-Dawley rats.
Test concentrations with justification for top dose:
Salmonella typhimurium Strain TA1538: 10 mg/ml
E. coli WP2 uvr A: 10 mg/L (Without S9) and 1, 5 or 10 mg/L (with S9)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Deionized water
- Justification for choice of solvent/vehicle: The test chemical was soluble in deionized water
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Deionized water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: Furacin (WP2uvrA, -S9), Rimmel Mahogany Silk (TA1538, -S9), 2-acetylaminofluorene (TA1538, +S9) and Tris-(2,3-dibromopropyl) phosphate (WP2uvrA, +S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: In medium (Fluctuation assay)

DURATION
- Preincubation period: No data
- Exposure duration: 72 hrs (without S9) or 96 hrs (with S9)
- Expression time (cells in growth medium): 72 hrs (without S9) or 96 hrs (with S9)
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available
NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available
Rationale for test conditions:
No data available
Evaluation criteria:
The tubes were scored for turbidity. When dyes such as Red 2G was used in this system, it may be impossible to detect the turbidity in the tubes by eye or to use a growth indicator such as bromothymol blue, due to masking by the color. In this case, the presence of viable prototrophic revertants was verified by streaking loopfuls from each tube onto non-supplemented agar.
Statistics:
No data available

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
No data available
Remarks on result:
other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:
Red 2G did not induce gene mutation in Salmonella typhimurium strain TA1538 and E. coli WP2 uvrA and hence is considered to be negative for gene mutation in vitro.
Executive summary:

In a mutagenicity test, the mutagenic effect of Red 2G was evaluated using Salmonella typhimurium strain TA1538 and E. coli WP2 uvrA by the Fluctuation test. The bacteria were exposed to the test compound at the concentration of 10 mg /ml in presence or absence of metabolic activation for strain TA1538 and 10 mg/L (Without S9) and 1, 5 or 10 mg/L (with S9) for strain WP2 uvrA. At the end of the study, the tubes were scored for turbidity. When dyes such as Red 2G was used in this system, it may be impossible to detect the turbidity in the tubes by eye or to use a growth indicator such as bromothymol blue, due to masking by the color. In this case, the presence of viable prototrophic revertants was verified by streaking loopfuls from each tube onto non-supplemented agar. Red 2G did not induce gene mutation in Salmonella typhimurium strain TA1538 and E. coli WP2 uvrA and hence is considered to be negative for gene mutation in vitro.