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Toxicological information

Genetic toxicity: in vitro

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in vitro gene mutation study in bacteria
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal

Data source

Reference Type:
Final Report on the Safety Assessment of Basic Blue 99
American College of Toxicology
Bibliographic source:
International Journal of Toxicology, 26(Suppl. 2):51–63, 2007

Materials and methods

Test guideline
according to
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

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.


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
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
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
No data available

Results and discussion

Test results
Species / strain:
E. coli, other: E. coli strain 343/113
Metabolic activation:
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
Migrated from field 'Test system'. Remarks: Bacterial tester strain used

Applicant's summary and conclusion

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.