Hexylamine

Genetic toxicity: in vitro

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

Test material information

Constituent 1

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.