Reaction products of diazotised sodium 2-amino-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate coupled with sodium 2-(4-aminobenzene-1-sulfonyl)ethyl sulfate, subsequently coupled with diazotised sodium 2-(4-aminobenzene-1-sulfonyl)ethyl sulfate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Remarks:

read across data

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:

Mutagenicity of azo dyes in the Salmonella/microsome assay using in vitro and in vivo activation

Author:

Fred Joachim , Ann Burrell , June Andersen

Year:

1985

Bibliographic source:

Mutation Research, 156 (1985) 131-138

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Salmonella/microsome assay was performed to evaluate the mutagenic response for the test chemical Food black 2 (FB2)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Tetrasodium 6-amino-4-hydroxy-3-[[7-sulphonato-4-[(4-sulphonatophenyl)azo]-1-naphthyl]azo]naphthalene-2,7-disulphonate

EC Number:

218-326-9

EC Name:

Tetrasodium 6-amino-4-hydroxy-3-[[7-sulphonato-4-[(4-sulphonatophenyl)azo]-1-naphthyl]azo]naphthalene-2,7-disulphonate

Cas Number:

2118-39-0

Molecular formula:

C26H15N5Na4O12S4

IUPAC Name:

Tetrasodium 6-amino-4-hydroxy-3-[[7-sulphonato-4-[(4-sulphonatophenyl)azo]-1-naphthyl]azo]naphthalene-2,7-disulphonate

Details on test material:

- Name of test material: Food black 2 (FB2)
- IUPAC name: tetrasodium (3E)-6-amino-4-oxo-3-(2-{7-sulfinato-4-[(E)-2-(4-sulfonatophenyl)diazen-1-yl] naphthalen-1-yl}hydrazin-1-ylidene)-3,4- dihydronaphthalene-2,7-disulfonate
- Molecular formula: C26H19N5O13S4.4Na
- Molecular weight: 809.651 g/mol
- Substance type: Organic

Specific details on test material used for the study:

- Name of test material: Food black 2 (FB2)
- IUPAC name: tetrasodium (3E)-6-amino-4-oxo-3-(2-{7-sulfinato-4-[(E)-2-(4-sulfonatophenyl)diazen-1-yl] naphthalen-1-yl}hydrazin-1-ylidene)-3,4- dihydronaphthalene-2,7-disulfonate
- Molecular formula: C26H19N5O13S4.4Na
- Molecular weight: 809.651 g/mol
- Substance type: Organic
- Physical state: No data available
- Purity: 85%
- Impurities (identity and concentrations): 15%

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1538 and TA98

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:

The S9 liver homogenate was prepared from rats treated with Aroclor 1254 5 days prior to S9 preparation.

Test concentrations with justification for top dose:

Doses upto 5 mg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

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 agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: No data
- Expression time (cells in growth medium): No data
- 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

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: 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

Evaluation criteria:

A dose-related increase (at least 2-fold) in revertant colonies was used to define a significant mutagenic response.

Statistics:

No data available

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1538 and TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

No data

Remarks on result:

other: No mutagenic potential

Applicant's summary and conclusion

Conclusions:

Food black 2 (FB2) did not induce reversion of mutation when applied to Salmonella typhimurium strains TA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

Salmonella/microsome assay was performed to evaluate the mutagenic response for the read across substance tetrasodium (3E)-6 -amino-4 -oxo-3 -(2 -{7 -sulfinato-4 -[(E)-2 -(4 -sulfonatophenyl)diazen-1 -yl]naphthalen-1 -yl}hydrazin-1 -ylidene)-3,4 -dihydronaphthalene-2,7 -disulfonate (CAS no. 2118 -39 -0, E.C. no. 218 -326 -9). The test was performed using Salmonella typhimurium strainsTA1538 and TA98 in the presence and absence of S9 metabolic activation system upto a dose level of 5 mg/plate. The test chemical was dissolved in DMSO and a dose-related increase (at least 2 -fold) in revertant colonies was used to define a significant mutagenic response. Food black 2 (FB2)did not induce reversion of mutation when applied to Salmonella typhimurium strainsTA1538 and TA98 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.