Dipotassium 4,4'-bis[6-anilino-4-[bis(2-hydroxyethyl)amino-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from publication.

Data source

Reference

Reference Type:

publication

Title:

Screening Of Food Dyes For Genotoxic Activity

Author:

R. B. Haveland-Smith And R. D. Combes

Year:

1979

Bibliographic source:

Fd. Cosmetic. Toxicol. Vol. 18, pp. 215 to 221

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

The induction of mutations was studied in modified liquid fluctuation tests using a tryptophan-requiring E. coli strain (sensitive to base substitutions) and a histidine auxotroph of Salmonella typhimurium (specific for frameshifts)for the test compound Patent Blue V.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen [4-[4-(diethylamino)-2',4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt

EC Number:

204-934-1

EC Name:

Hydrogen [4-[4-(diethylamino)-2',4'-disulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene]diethylammonium, sodium salt

Cas Number:

129-17-9

Molecular formula:

C27H32N2O6S2.Na

IUPAC Name:

sodium 4-{[4-(diethylamino)phenyl][4-(diethyliminio)cyclohexa-2,5-dien-1-ylidene]methyl}benzene-1,3-disulfonate

Details on test material:

- Name of test material: Patent Blue V
- Molecular formula: C27H31N2O6S2.Na
- Molecular weight: 566.672 g/mol
- Substance type: Organic
- Physical state: Solid
Purity: No data available
- Impurities: No data available

Specific details on test material used for the study:

- Name of test material: Patent Blue V
- Molecular formula: C27H31N2O6S2.Na
- Molecular weight: 566.672 g/mol
- Substance type: Organic
- Physical state: Solid

Method

Target gene:

Tryptophan- E. coli strain
Histidine auxotroph of Salmonella typhimurium

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:

not specified

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):

not specified

Metabolic activation:

with and without

Metabolic activation system:

rat-liver microsomal activation.

Test concentrations with justification for top dose:

0.5 mg/ml

Vehicle / solvent:

Deionised water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Deionised water

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: 72-96 hrs
- Expression time (cells in growth medium): 72-96 hrs
- 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: Triplicate

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: The food colour was made up in deionized water and membrane-sterilized prior to use. The test material was tested at its maximum sublethal concentration.

Evaluation criteria:

The test material was considered positive only if it resulted in significant more turbid tubes in a treated series when compared with an untreated set of tubes

Statistics:

Chi square test

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:

not specified

Positive controls validity:

not specified

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

Remarks on result:

other: No mutagenic effect were observed.

Applicant's summary and conclusion

Conclusions:

Patent blue V (129-17-9) was evaluated for its mutagenic potential in bacteria.The test substance was failed to induce gene mutation in the bacterial tester strains Salmonella typhimurium TA 1538 and Escherichia coli WP2 uvrA.

Executive summary:

Patent blue V was assessed for its possible mutagenic potential. For this purpose fluctuation test was performed used at a concentration of 0.5 mg/ml in liquid medium for 72-96 hrs. The tester strains used were Salmonella typhimurium TA 1538 and Escherichia coli WP2 uvrA. The assay was performed for each bacterium in three separate experiments.

The given test material failed to induce genotoxicity in the bacteria Salmonella typhimurium TA 1538 and Escherichia coli WP2 uvrA. Therefore Patent blue V was considered to non mutagenic .Hence does not classify for gene mutation in vitro.