4-[(3-aminobenzoyl)amino]-5-hydroxynaphthalene-1,7-disulphonic acid

Genetic toxicity: in vitro

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

Test material information

Constituent 1

Reference substance name:

Red 2G

IUPAC Name:

Red 2G

Constituent 2

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 / strain

Species / strain 1

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 2

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 results

Test results 1

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

Test results 2

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.