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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIV)-in vitro Chromosomal Aberration Assay with 11 Chemicals in Chinese Hamster Lung Cells.
Author:
Youn-Jung Kim & Jae-Chun Ryu
Year:
2006
Bibliographic source:
MOLECULAR & CELLULAR TOXICOLOGY, Vol. 2, No. 2, 89-96, June 2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Principles of method if other than guideline:
Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIV)-in vitro Chromosomal Aberration Assay with 11 Chemicals in Chinese Hamster Lung Cells. One of the chemicals studied include is n-Hexylamine
GLP compliance:
not specified
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Constituent 1
Chemical structure
Reference substance name:
Hexylamine
EC Number:
203-851-8
EC Name:
Hexylamine
Cas Number:
111-26-2
Molecular formula:
C6H15N
IUPAC Name:
hexan-1-amine
Test material form:
other: liquid
Details on test material:
- Name of test material: Hexylamine
- Molecular formula: C6H15N
- Molecular weight:101.19 g/mol
- Smiles notation (if other than submission substance): C(CCC)CCN
- InChl: 1S/C6H15N/c1-2-3-4-5-6-7/h2-7H2,1H3
- Substance type: Organic
- Physical state: liquid

Method

Target gene:
Chinese Hamster Lung Cells
Species / strain
Species / strain / cell type:
other: Chinese Hamster Lung Cells
Details on mammalian cell type (if applicable):
- Type and identity of media: Monolayer media with EMEM supplemented with 10% FBS, 50 units/mL penicillin and 50 μg/mL streptomycin.
- Properly maintained: Yes, maintained at 370 C in humidified 5% CO2 atmosphere.

Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 activation
Test concentrations with justification for top dose:
36.3, 72.5, 144.9, 160.3, 320.5 and 640.9 μg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Dimethylsulfoxide (DMSO) were used.
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: Cyclophosphamide and mitomycin C (MMC)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation; in suspension; as impregnation on paper disk: Dose were prepared and separately added to 3-day-old Eagles minimum essential medium (EMEM) cultures (approximately 105 cells/60 mm dish).
DURATION
- Preincubation period: No data available
- Exposure duration: 6 hr and 24 hr
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent): 22 hr followed by addition of medium containing colcemid
- Fixation time (start of exposure up to fixation or harvest of cells): 2hr

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: few drop of cell were evaluated for chromosomal aberrations.

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Yes,
For dose range-finding study, study performed in the absence of a rat liver S9 activation system. For the growth inhibition assay, CHL cells were seeded at the density of 5×104 cells/mL into 96 well plates. 24 hr after seeding, several different doses of sample were separately added and incubated for 6 hr in the presence of S9 activation system and 24 hr in the absence of S9 system. And then the 50% inhibition concentration (IC50) values were calculated by MTT assay.

OTHER EXAMINATIONS:
- Determination of polyploidy: Yes
- Determination of endoreplication: Yes
- Other: No data available

OTHER: No data available
Evaluation criteria:
Absence and presence of chromosome aberrations
Statistics:
Data were analyzed by using Fishers exact test48 with Dunnetts adjustment and compared with results from the solvent controls.
Data from count up well-spread 200 metaphase cells were expressed as percentages, and then dose-dependent responses and the statistical significance in p-value will be considered as positive results.

Results and discussion

Test results
Species / strain:
other: Chinese Hamster Lung Cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: Dose finding study, result not mention
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES: Yes, performed in the absence of a rat liver S9 activation system. For the growth inhibition assay, CHL cells were seeded at the density of 5×104 cells/mL into 96 well plates. 24 hr after seeding, several different doses of sample were separately added and incubated for 6 hr in the presence of S9 activation system and 24 hr in the absence of S9 system. And then the 50% inhibition concentration (IC50) values were calculated by MTT assay.


Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and without

n-Hexylamine in the concentration of 36.3, 72.5, 144.9, 160.3, 320.5 and 640.9 μg/mL did not show any evidence of gene toxicity when CHL cells were exposed to the test chemical and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

In a gene toxicity test, Chinese Hamster Lung Cells (CHL) cells were exposed to n-Hexylamine at a concentration of 36.3, 72.5, 144.9, 160.3, 320.5 and 640.9 μg/mL with and without metabolic activation for 6 and 24 hours. The results showed that there was no significant evidence of chromosome aberrations after treatment. Independently of tested n-Hexylamine concentration, the results showed no evidence of gene toxicity. Therefore, it is considered that n-Hexylamine in the concentration of 144.9 μg/mL without S9 and 640.9 μg/mL with S9 does not cause genetic chromosome aberrations when CHL cells are exposed to the test chemical in the absence and inpresence of S9 activation system of S9 system for 6 and 24 hr. Hence the test chemical is not liekly to classify as a gene mutant in vitro.