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Toxicological information

Genetic toxicity: in vivo

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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:
1990
Bibliographic source:
Teratogenesis, Carcinogenesis, and Mutagenesis 10:449-462 (1990)

Materials and methods

Test guideline
Qualifier:
according to
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
Name:
Unnamed
Type:
Constituent
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
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 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.