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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The substance n-Hexylamine was found to be non mutagenic in in-vitro study.

Link to relevant study records
Reference
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
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
Target gene:
Chinese Hamster Lung Cells
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.
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.

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'.
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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Gene toxicity in vitro:

Data from reliable publication for the target chemical and its read aross have been summarized below to determine the mutagenic nature of the test compound Hexylamine:

In a gene toxicity test, Chinese Hamster Lung Cells (CHL) cells were exposed ton-Hexylamine at aconcentration 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 testedn-Hexylamineconcentration, the results showed no evidence of gene toxicity. Therefore, it is considered thatn-Hexylaminein 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.

Another gene mutation toxicity study was performed for the test chemical n- hexylamine to evaluate its mutagenic nature (U. S. Department of Health and Human Services, 2016). The AMES assay was conducted on Salmonella typhimurium strains TA 97, TA 98, TA 1535 and TA100. 10% and 30% RLI: induced male Sprague Dawley rat liver S9; HLI : induced male Syrian hamster liver S9 were used for metabolic activation. The assays were carried out according to the Pre Incubation method. Individual strain data is presented as mean ± standard error. n hexylamine exhibited no mutagenic activity with and without metabolic activation at all test concentrations.

In a study on read across (RA CAS no 110 -58 -7), the test chemical n-amylamine was tested (Zeiger et al, 1986) using a pre-incubation modification of the Salmonella microsome test in the absence of exogenous metabolic activation and in the presence of liver S-9 from Aroclor-induced male Sprague-Dawley rats and Syrian hamsters to evaluate its mutagenic nature. The chemical was tested in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and TA97 both in the presence and absence of S9 metabolic activation system. Concurrent solvent and positive controls were also used. The test chemical n-amylamine failed to induce mutation in the Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and TA97 in the presence and absence of S9 metabolic activation system and hence is not likely to classify for gene mutation in vitro.

Based on the data summarized, the test chemical hexylamine is not likely to classify as a gene mutant in vitro.

Justification for selection of genetic toxicity endpoint

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.

Justification for classification or non-classification

Based on the data summarized, the test chemical hexylamine is not likely to classify as a gene mutant in vitro.