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EC number: 201-635-8 | CAS number: 85-83-6
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Remarks:
- Similar to OECD 471
- Justification for type of information:
- Data is from peer reviewed publication
Data source
Reference
- Reference Type:
- publication
- Title:
- Mutagenicity Testing of Certified Food Colors and Related Azo, Xanthene And Triphenylmethane Dyes with the Salmonella/Microsome System
- Author:
- Joseph P. Brown
- Year:
- 1 978
- Bibliographic source:
- Mutation Research
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: Similar to OECD 471
- Principles of method if other than guideline:
- Ames assay was performed to determine the mutagenic nature of Test chemical .
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 1-(2-methyl-4-(2-methylphenylazo)phenylazo)-2-naphthol
- EC Number:
- 201-635-8
- EC Name:
- 1-(2-methyl-4-(2-methylphenylazo)phenylazo)-2-naphthol
- Cas Number:
- 85-83-6
- Molecular formula:
- C24H20N4O
- IUPAC Name:
- 1-(2-methyl-4-(2-methylphenylazo)phenylazo)-2-naphthol
- Details on test material:
- - Name of test material: Sudan IV
- Molecular formula: C24H20N4O
- Molecular weight: 380.449 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data
- Smiles: c12c(\N=N\c3c(cc(\N=N\c4c(cccc4)C)cc3)C)c(ccc1cccc2)O
Constituent 1
Method
- Target gene:
- Histidine
Species / strain
- Species / strain / cell type:
- S. typhimurium, other: TA1535, TA100, TA1537, TA1538, 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 mammalian liver homogenate fraction "S-9" was prepared from the liver of female Sprague Dawley rats injected i.p. with about 0.5 ml of a corn oil solution of Aroclor 1254 (400 mg/ml) to give a dose of 500 mg/kg body weight.
- Test concentrations with justification for top dose:
- 100 or 500 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test materail was soluble in DMSO
Controls
- Untreated negative controls:
- yes
- Remarks:
- Sodium dithionite
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- ethylmethanesulphonate
- methylmethanesulfonate
- other: Anthragallol and 2-Anthramine
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: No data
- Exposure duration: 3 days
- Expression time (cells in growth medium): 3 days
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data
NUMBER OF REPLICATIONS: No data
METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: No data
NUMBER OF CELLS EVALUATED: No data
NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): No data
CRITERIA FOR MICRONUCLEUS IDENTIFICATION: No data
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:
- Any supplementary information relevant to cytotoxicity: No data
OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable): No data
- OTHER: No data - Rationale for test conditions:
- No data
- Evaluation criteria:
- The criteria adopted for scoring a mutagenic response in routine plate tests was the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on historical controls.
- Statistics:
- No data
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium, other: TA1535, TA100, TA1537, TA1538 and TA98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- Without reduction performed using Sodium dithionite
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- For TA1538 at 500 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium, other: TA1535, TA100, TA1537, TA1538 and TA98
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Remarks:
- With reduction by sodium dithionite at 100 µg/plate
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium, other: TA1535 and TA100
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Remarks:
- With reduction performed using sodium dithionite at 100 µg/plate
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium, other: TA1537, TA1538 and TA98
- Metabolic activation:
- with
- Genotoxicity:
- positive
- Remarks:
- With reduction performed using Sodium dithionite at 100 µg/plate
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- The chemical reduction of Sudan IV was performed by dissolving the chemical in DMSO with an equal volume of freshly prepared aqueous dithionite solution of the same weight concentration.
- Remarks on result:
- other: No mutagenic effect were observed
Any other information on results incl. tables
Table: Screening of Sudan IV
Test concentration |
Chemical reduction (dithionite) |
Microsome activation |
Number of His+revertants/plate |
||||
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
|||
500µg/plate |
- |
- |
46 |
80 |
8 |
12* |
22 |
|
|
+ |
35 |
115 |
14 |
48 |
50 |
100µg/plate |
+ |
- |
24 |
110 |
10 |
23 |
19 |
|
|
+ |
17 |
105 |
35 |
303 |
400 |
*: Microbial toxicity
Applicant's summary and conclusion
- Conclusions:
- Test chemical failed to induce mutation in Salmonella typhimurium strain TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system without reduction using sodium dithionite at 500 µg/plate and without S9 activation in the same strains with reduction being performed at 100 µg/plate and also in strains TA1535 and TA100 in the presence of S9 metabolic activation system with reduction at 100 µg/plate. It however induced mutation in the strains TA1537, TA1538 and TA98 in the presence of S9 metabolic activation system with reduction at 100 µg/plate. Based on the results of the study, Sudan IV does not induce mutation without reduction being performed using sodium dithionite in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
- Executive summary:
Ames assay was performed to determine the mutagenic nature of test chemical in the presence and absence of S9 metabolic activation system. The study was also performed to determine the mutagenic nature of component amines formed upon chemical reduction. Non water soluble Sudan IV was dissolved in DMSO ans mixed with an equal volume of freshly prepared aqueous dithionite solution of the same weight concentration for reduction. The plates were incubated at 37 °C for 3 days prior to determining the number of revertants/plate. The criteria adopted for scoring a mutagenic response in routine plate tests was that the observed number of revertants should exceed twice the background value for that given assay and exceed the 99.9% confidence limit based on historical controls. Test chemical failed to induce mutation in Salmonella typhimurium strain TA1535, TA100, TA1537, TA1538 and TA98 in the presence and absence of S9 metabolic activation system without reduction using sodium dithionite at 500 µg/plate and without S9 activation in the same strains with reduction being performed at 100 µg/plate and also in strains TA1535 and TA100 in the presence of S9 metabolic activation system with reduction at 100 µg/plate. It however induced mutation in the strains TA1537, TA1538 and TA98 in the presence of S9 metabolic activation system with reduction at 100 µg/plate. Based on the results of the study, test chemical does not induce mutation without reduction being performed using sodium dithionite in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
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