Dodecanenitrile

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Evaluation of genotoxicity of nitrile fragrance ingredients using in vitro and in vivo assays

Author:

S.P. Bhatia, V.T. Politano, A.M. Api

Year:

2013

Bibliographic source:

Food and Chemical Toxicology 59 (2013) 784–792

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

not specified

Type of assay:

other: In vitro chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Dodecanenitrile

EC Number:

219-440-1

EC Name:

Dodecanenitrile

Cas Number:

2437-25-4

Molecular formula:

C12H23N

IUPAC Name:

dodecanenitrile

Details on test material:

SMILES:CCCCCCCCCCCC#N

Specific details on test material used for the study:

- Name of test material (IUPAC name): Dodecanenitrile
- Molecular formula: C12H23N
- Molecular weight: 181.321 g/mole
- Smiles notation: C(CCCCCC)CCCCC#N
- InChl: 1S/C12H23N/c1-2-3-4-5-6-7-8-9-10-11-12-13/h2-11H2,1H3
- Substance type: Organic
- Physical state: colourless liquid

Method

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9 fractions

Test concentrations with justification for top dose:

without S9: 7.2, 14.5 and 28.9 µg/ml
with S9: 426.5, 925, 1850 µg/ml
A preliminary toxicity assay was performed to determine dose selection for the cytogenetic experiments.

Vehicle / solvent:

DMSO 0.5% (v/v)

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

cyclophosphamide

ethylmethanesulphonate

Details on test system and experimental conditions:

A preliminary toxicity assay was performed to determine dose selection for the cytogenetic experiments. In each experimental
group two parallel cultures were set up with and without metabolic activation using rat liver S9 fractions. 100 Metaphase plates per culture were scored for structural chromosome aberrations. The exposure period was 4 h.

Rationale for test conditions:

not specified

Evaluation criteria:

100 Metaphase plates per culture were scored for structural chromosome aberrations

Statistics:

Fisher’s exact test (p < 0.05) was used to determine statistical significance.

Results and discussion

Test results

Key result

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No biologically relevant increase in the frequencies of polyploid metaphases was observed after treatment with the test material as compared to the frequencies of the controls.

Any other information on results incl. tables

Summary of in vitro chromosome aberration test with dodecanenitrile:

Preparation interval	Test item concentration in lg/mL	Polyploid cells in %	Cell no in % of control	Mitotic indices in % of control	Including gaps^a	Aberrant cells in % excluding gaps^a	With exchanges
Exposure period 4-h without S9
18-h	DMSO 0.5% (v/v)	2.4	100.0	100.0	0.5	0.5	0.5
	EMS 900µg/ml	3.5	n.t.	102.7	11.5	11.5^s	35
	7.2	3.6	102.2	91.2	0.5	0.0	0.0
	14.5	3.1	80.1	89.7	0.5	0.0	0.0
	28.9	2.6	62.9	108.0	1.5	1.0	0.5
Exposure period 4-h with S9
18-h	DMSO 0.5% (v/v)	1.6	100.0	100.0	1.5	1.0	0.5
	CPA 1.4µg/ml	2.9	n.t.	95.8	14.5	13.5^s	6.5
	426.5	3.7	93.4	82.7	3.0	3.0	0.5
	925.0	3.7	100.0	94.3	3.0	2.0	0.5
	1850	3.2	83.9	96.1	1.0	1.0	0.5

n.t. not tested.
^aInclusive cells carrying exchanges.
^b100 Metaphase plates per culture were evaluated.
^c50 Metaphase plates per culture were evaluated.
^sSignificantly higher.

Applicant's summary and conclusion

Conclusions:

Dodecanenitrile was determined to be non-clastogenic in an in vitro chromosome aberration assay in V79 cells with and without metabolic activation using rat liver S9 fractions.

Executive summary:

In vitro chromosome aberration assays were performed using Chinese hamster V79 cells according to OECD Testing Guideline No. 473 and were conducted on dodecanenitrile (CAS 2437-25-4). A preliminary toxicity assay was performed to determine dose selection for the cytogenetic experiments The chromosomes were prepared 18 h after start of treatment with the test item. V79 cells were exposed to test material dissolved in DMSO, as well as positive and solvent control ±S9 activation. The exposure period was 4 h (±S9). In experimental group two parallel cultures were set up. Per culture at least 100 metaphase plates were scored for structural chromosome aberrations were scored. The positive control without metabolic activation was ethyl methane sulfonate (EMS) and with metabolic activation was cyclophosphamide (CPA). The positive controls induced statistically significant increases (p < 0.05) in cells with structural chromosome aberrations. Dodecanenitrile was determined to be non-clastogenic in an in vitro chromosome aberration assay in V79 cells with and without metabolic activation using rat liver S9 fractions.

Based on the above result, it can be concluded that the substance dodecanenitrile does not present a concern for genotoxic potential and can be considered as not classified as per CLP classification criteria.