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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)

Data source

Reference Type:
A Compilation of Two Decades of Mutagenicity Test Results with the Ames Salmonella typhimurium and L5178Y Mouse Lymphoma Cell Mutation Assays.
H. E. Seifried, R. M. Seifried, J. J. Clarke, T. B. Junghans, and R. H. C. San
Bibliographic source:
Chem. Res. Toxicol., Vol. 19 (5), Pg. no. 627–644, 2006

Materials and methods

Test guideline
according to guideline
other: as mentioned below
Principles of method if other than guideline:
Bacterial gene mutation assay was carried out using S. typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Constituent 1
Reference substance name:
Brilliant Black 1
Brilliant Black 1
Constituent 2
Chemical structure
Reference substance name:
Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate
EC Number:
EC Name:
Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate
Cas Number:
Molecular formula:
tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate
Details on test material:
- Name of test material (as cited in study report): C.I Briliant black BN- Molecular formula (if other than submission substance): C28-H21-N5-O14-S4.4Na- Molecular weight (if other than submission substance): 867.6873- Substance type: Organic- Physical state: Solid- Purity: No data available- Impurities (identity and concentrations): No data available


Target gene:
Plate incorporation method
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA98, TA100, TA1535, TA1537, and TA1538.
Details on mammalian cell type (if applicable):
No data available
Additional strain / cell type characteristics:
other: Salmonella typhimurium is a histidine auxotroph strain.
Metabolic activation:
with and without
Test concentrations with justification for top dose:
Test concentration: Dose (µg /plate): conc. in the range of 10-10000 µg /plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Appropriate solvent was used for preparing the stock solution of the test chemical. Name of the solvent is not specified in the study.
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
not specified
True negative controls:
Positive controls:
Positive control substance:
not specified
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporationDURATION- Preincubation period: 48-h- Exposure duration: 37 ± 2ᵒC- Expression time (cells in growth medium): 48 hr- 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 availableSELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: TriplicatesNUMBER OF CELLS EVALUATED: 1-2 × 109 cells/mlDETERMINATION OF CYTOTOXICITY- Method: No data available- Other: Cytotoxicity was observed in a preliminary dose range-finding study using strain TA100.OTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication: No data available- Other: : No data availableOTHER: The test materials were stored at -20ᵒC, 4ᵒC, or room temperature as required to maintain compound stability. The test chemical was analyzed for purity by Midwest Research Institute (MRI).Five doses of test chemical, together with the appropriate concurrent solvent and positive controls, were tested on each tester strain without metabolic activation and also with activation by induced rat and hamster liver S9 preparations. The test bacterial cultures were grown overnight in Oxoid nutrient broth no. 2 and were removed from incubation when they reached a density of (1-2) × 109 cells/ml.On the day of use, all tester strain cultures were checked for genetic integrity.
Evaluation criteria:
The result was considered positive if at least a doubling in the mean number of revertants/plate of at least one tester strain was found.
No data available

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA98, TA100, TA1535, TA1537, and TA1538.
Metabolic activation:
with and without
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data available- Effects of osmolality: No data available- Evaporation from medium: No data available- Water solubility: No data available- Precipitation: No data available- Other confounding effects: No data availableRANGE-FINDING/SCREENING STUDIES: Preliminary dose range-finding study was performed using Salmonella typhimurium strain TA100.COMPARISON WITH HISTORICAL CONTROL DATA: No data availableADDITIONAL INFORMATION ON CYTOTOXICITY: No data available
Remarks on result:
other: other: Bacterial tester strain
Migrated from field 'Test system'.

Applicant's summary and conclusion

Interpretation of results (migrated information):negative Negative (with and without)The test substance C.I Brilliant black BN ( food black 1) had shown non-mutagenic response in the test bacterial strain TA98, TA100, TA1535, TA1537, and TA1538. (with and without metabolic activation system).
Executive summary:

The mutagenic potency ofC.I Brilliant black BN( food black 1) was tested by the plate incorporation method usingSalmonella typhimuriumstrainTA98, TA100, TA1535, TA1537, and TA1538. When the test bacterial strain is exposed with the test chemical for 48hrs, no mutagenic response was seen in any of the strains of Salmonella typhimurium(with and without metabolic activation system).