3-[(4-amino-6-bromo-5,8-dihydro-1-hydroxy-8-imino-5-oxo-2-naphtyl)amino]-N,N,N-trimethylanilinium chloride

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:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Final Report on the Safety Assessment of Basic Blue 99

Author:

American College of Toxicology

Year:

2007

Bibliographic source:

International Journal of Toxicology, 26(Suppl. 2):51–63, 2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Bacterial mutagenicity assay was carried out on E. coli strain 343/113 to test the genotoxic potential of Basic Blue 99.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-[(4-amino-6-bromo-5,8-dihydro-1-hydroxy-8-imino-5-oxo-2-naphtyl)amino]-N,N,N-trimethylanilinium chloride

EC Number:

268-544-3

EC Name:

3-[(4-amino-6-bromo-5,8-dihydro-1-hydroxy-8-imino-5-oxo-2-naphtyl)amino]-N,N,N-trimethylanilinium chloride

Cas Number:

68123-13-7

Molecular formula:

C19H20BrN4O2.Cl

IUPAC Name:

3-[(4-amino-6-bromo-5,8-dihydro-1-hydroxy-8-imino-5-oxo-2-naphtyl)amino]-N,N,N-trimethylanilinium chloride

Constituent 2

Reference substance name:

Basic Blue 99

IUPAC Name:

Basic Blue 99

Details on test material:

Details on test material- Name of test material (as cited in study report): Basic Blue 99- Molecular formula (if other than submission substance): C19-H20-Br-N4-O2.Cl- Molecular weight (if other than submission substance): 451.75 g/mol- Substance type: Organic- Physical state: Solid- Purity According to the Keystone Aniline Corporation (1999) the chemical is atleast 63% pure Another supplier of Basic Blue 99 (Henkel 1992) has a specification of 60.7% dye content- Impurities (identity and concentrations): According to the Keystone Aniline Corporation (1999) have no more than 100 ppm of ironAnother supplier of Basic Blue 99 (Henkel 1992) suggests the presence of 25.7% sugar content, 11.8% inorganic salts, and 1.8 % volatile matter/water crystallization.

Method

Target gene:

E. coli strain 343/113

Species / strain

Species / strain / cell type:

E. coli, other: E. coli strain 343/113

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

No data

Test concentrations with justification for top dose:

1, 10, and 100 μg/ml

Vehicle / solvent:

No data

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Remarks:

no data

Details on test system and experimental conditions:

Details on test system and conditionsMETHOD OF APPLICATION: preincubation- in agar (plate incorporation)DURATION- Preincubation period: 2 hrs- Exposure duration: 2 hrs- Expression time (cells in growth medium): 20-72 hrs- Selection time (if incubation with a selection agent): Selective medium was used for bacterial expression - Fixation time (start of exposure up to fixation or harvest of cells):SELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: No data availableNUMBER OF CELLS EVALUATED: No data availableDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No data availableOTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication : No data available- Other: No data availableOTHER: No data available

Evaluation criteria:

Reversion of mutation was observed by the couting the mutant colonies

Statistics:

No data available

Results and discussion

Test results

Species / strain:

E. coli, other: E. coli strain 343/113

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: Bacterial tester strain used

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):negative without metabolic activationBasic blue 99 is non mutagenic in the E. coli strain 343/113.

Executive summary:

Bacterial mutagenicity assay was carried out onE. colistrain 343/113 to test the genotoxic potential of Basic Blue 99. The test compound was tested at the concentrations of 1, 10, and 100μg/ml and a volume of 0.1 ml in a dark place at 37⁰C for 2 h. Cells were spread over four selected media and incubated for 20 to 72 h. Cells were counted and analyzed for reverse mutations of arg− to arg+ and nad− to nad+,

forward mutations of 5-methyl-dl-trytophan-sensitivity to MT resistance, and forward and reverse mutations of gal Rs 18 to gal+. The mutant colonies were counted and compared with controls.

Basic blue 99 is non mutagenic in theE. colistrain 343/113.

According to the CLP classification, the test material Basic Blue 99 does not classify as a mutagen in vitro.