Registration Dossier
Registration Dossier
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EC number: 208-953-6 | CAS number: 548-62-9
Genetic toxicity: in vivo
Administrative data
- Endpoint:
- in vivo mammalian cell study: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer reviewed publication
Data source
Reference
- Reference Type:
- publication
- Title:
- Evaluation of the Genotoxicity of Gentian Violet in Bacterial and Mammalian Cell Systems
- Author:
- Anane Aidoo, Ning Gao, Robin E. Neft, Henry M. Schol, Bruce S. Hass, Toni Y. Minor, and Robert H. Heflich
- Year:
- 1 990
- Bibliographic source:
- Teratogenesis, Carcinogenesis, and Mutagenesis 10:449-462 (1990)
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- Nucleoid Sedimentation Analysis was performed in vivo to determine the DNA damaging effect of Gentian violet
- GLP compliance:
- not specified
- Type of assay:
- other: Nucleoid Sedimentation Analysis Assay
Test material
- Reference substance name:
- [4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride
- EC Number:
- 208-953-6
- EC Name:
- [4-[4,4'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium chloride
- Cas Number:
- 548-62-9
- Molecular formula:
- C25H30ClN3
- IUPAC Name:
- 4-{bis[4-(dimethylamino)phenyl]methylidene}-N,N-dimethylcyclohexa-2,5-dien-1-iminium chloride
- Test material form:
- solid
- Details on test material:
- - Name of test material (as cited in study report):Gentian violet
- IUPAC name: N-(4-{bis[4-(dimethylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium chloride
- Molecular formula : C25H30N3.Cl
Molecular weight : 407.986 g/mol
- Smiles notation: C(\c1ccc(N(C)C)cc1)(c1ccc(N(C)C)cc1)=C1\C=C\C(=[N+](/C)C)C=C1.[ClH-]
- InChl:1S/C25H30N3.ClH/c1-26(2)22-13-7-19(8-14-22)25(20-9-15-23(16-10-20)27(3)4)21-11-17-24(18-12-21)28(5)6;/h7-18H,1-6H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
Constituent 1
- Specific details on test material used for the study:
- - Name of test material: Gentian violet
- IUPAC name: 4-{bis[4-(dimethylamino)phenyl]methylidene}-N,N-dimethylcyclohexa-2,5-dien-1-iminium chloride
- Molecular formula: C25H30ClN3
- Molecular weight: 407.986 g/mol
- Substance type: Organic
- Physical state:
- Purity: 97% dye
- Impurities (identity and concentrations): 3 %
Test animals
- Species:
- mouse
- Strain:
- B6C3F1
- Details on species / strain selection:
- No data
- Sex:
- not specified
- Details on test animals or test system and environmental conditions:
- No data
Administration / exposure
- Route of administration:
- intravenous
- Vehicle:
- Vehicles
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO
- Concentration of test material in vehicle: 0- 10 µg/mL
- Amount of vehicle (if gavage or dermal): No data
- Type and concentration of dispersant aid (if powder): No data
- Lot/batch no. (if required): No data
- Purity: No data - Details on exposure:
- Not applicable
- Duration of treatment / exposure:
- Duration of exposure: 1 hour
- Frequency of treatment:
- No data
- Post exposure period:
- No data
Doses / concentrations
- Remarks:
- 10 µg/mL
- No. of animals per sex per dose:
- No data
- Control animals:
- yes
- Positive control(s):
- Methylmethanesulphonate
- Route of administration: No data
- Doses / concentrations: No data
Examinations
- Tissues and cell types examined:
- Spleen lymphocytes
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: No data
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): No data
DETAILS OF SLIDE PREPARATION: No data
METHOD OF ANALYSIS: The position of nucleoids generated was analyzed
OTHER: Cells were washed with PBS and resuspended in PBS at a concentration of 2 to 10 X 106 cells/ml. A 50 µL sample was then added to 150 µL of lysing solution (2.5 M NaCl, 0.133 M EDTA, 2.67 mM Tris, and 0.67% Triton X-100, pH 8.0) layered on top of a 15-30% neutral (pH 8.0) sucrose gradient containing 1.95 M NaCl, 0.01 M Tris, 0.001 M EDTA, and 0.1 kg/ml Hoechst 33258 dye. Following a 30-minute lysis in the dark at room temperature, the gradients were centrifuged at 25,000 rpm for 75 minutes at 20°C in an SW41 rotor. Six gradients were run per rotor with one gradient serving as a reference. The position of the nucleoids in the gradients was visually determined with a UV light. - Evaluation criteria:
- The position of nucleoids generated was analyzed and results were expressed as the ratio of the distance traveled by nucleoids in the sample gradients to the reference gradient
- Statistics:
- No data
Results and discussion
Test results
- Sex:
- not specified
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- Concentration more than 10 µg/mL were fatal to the mice during the 1 hour treatment period
- Vehicle controls validity:
- not specified
- Negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- No data
Applicant's summary and conclusion
- Conclusions:
- No DNA damage was detected by nucleoid sedimentation analysis of lymphocytes isolated from B6C3F1 mice exposed to different concentrations of Gentian Violet.
- Executive summary:
Nucleoid Sedimentation Analysis was performed in vivo to determine the DNA damaging effect of Gentian violet. The study was performed using B6C3F1mice. The animals were pretreated for 1 hour with 0- 10µg/mL Gentian Violet administered by injection into the tail-vein. The spleen lymphocytes were isolated from the treated animals by lympho-plaque centrifugation. Animals treated with MMS were used as positive controls.The position of nucleoids generated was analyzed and results were expressed as the ratio of the distance traveled by nucleoids in the sample gradients to the reference gradient. No DNA damage was detected by nucleoid sedimentation analysis of lymphocytes isolated from B6C3F1 mice exposed to different concentrations of Gentian Violet and hence the test chemical does not exhibit gene mutation in vivo.
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