Cinnamonitrile

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / micronucleus study

Type of information:

experimental study

Adequacy of study:

weight of evidence

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 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 487 (In vitro Mammalian Cell Micronucleus Test)

Principles of method if other than guideline:

In vitro micronucleus test was performed to determine the mutagenic nature of cinnamyl nitrile

GLP compliance:

not specified

Type of assay:

other: In vitro micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

Cinnamonitrile

EC Number:

217-552-5

EC Name:

Cinnamonitrile

Cas Number:

1885-38-7

Molecular formula:

C9H7N

IUPAC Name:

cinnamonitrile

Details on test material:

- Name of test material (IUPAC name): (E)-3-phenylprop-2-enenitrile
- Common name: Cinnamyl nitrile
- Molecular formula: C9H7N
- Molecular weight: 129.161 g/mol
- Smiles notation: N#C\C=C\c1ccccc1
- InChl: 1S/C9H7N/c10-8-4-7-9-5-2-1-3-6-9/h1-7H/b7-4+
- Substance type: Organic

Specific details on test material used for the study:

- Name of test material: Cinnamyl nitrile
- IUPAC name: (E)-3-phenylprop-2-enenitrile
- Molecular formula: C9H7N
- Molecular weight: 129.161 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Method

Target gene:

No data

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

No data

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

Liver S9 mix was isolated from phenobarbital/-naphthoflavone treated rats

Test concentrations with justification for top dose:

2.5, 5.1, 10.2, 20.3, 40.6, 81.3, 162.5, 325, 650, 1300 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

other: Colcemid (7.5 and 10.0 µg/mL)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: No data
- Exposure duration: 4 hrs
- Expression time (cells in growth medium): 24 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

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

NUMBER OF REPLICATIONS: No data

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: cell count was measured

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

OTHER: All cultures were incubated at 37˚C in a humidified atmosphere with 1.5% CO2 (98.5% air).

Rationale for test conditions:

No data

Evaluation criteria:

The cells were observed for the presence of micronuclei

Statistics:

Statistical analysis were conducted using Chi square test, however biological relevant factors such as dose dependency and historical data were considered along with statistical significance

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

False positive / 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 data

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Summary of in vitro micronucleus test

Compound	Preparation interval (h)	Concentrations tested (µg/mL)	Concentrations scored (µg/mL)	Cell count (%)	Micronuclei (%)
Exposure period of 4 h with S9 mix Cinnamyl nitrile	24	2.5, 5.1, 10.2, 20.3, 40.6, 81.3, 162.5, 325, 650, 1300	162.5	86.4	0.90
			325	44.2	6.10^S
			650^P	3.8	3.60^P
Exposure period of 4 h withot S9 mix Cinnamyl nitrile	24	2.5, 5.1, 10.2, 20.3, 40.6, 81.3, 162.5, 325, 650, 1300	40.6	59.4	16.50^S
			81.3	44.4	17.85^S
			162.5	38.7	24.55^S

P: Precipitation occurred.

S: Frequency statistically significant higher than corresponding control values.

Applicant's summary and conclusion

Conclusions:

Cinnamyl nitrile induced micronuclei formation in Chinese Hamster V79 cells in the presence and absence of S9 metabolic activation system. However the data obtained in this study provides false positive results and hence the test chemical is considered to be negative for gene mutation in vitro.

Executive summary:

In vitro micronucleus test was performed to determine the mutagenic nature of cinnamyl nitrile. The study was performed as per OECD 487 using Chinese Hamster V79 cellsin the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose levels of 0,2.5, 5.1, 10.2, 20.3, 40.6, 81.3, 162.5, 325, 650, 1300µg/mL. The cells were exposed to the test chemical for 4 hours and were provided with an expression time of 24 hrs.Cinnamyl nitrile induced micronuclei formation in Chinese Hamster V79 cells in the presence and absence of S9 metabolic activation system.

 

However the data obtained in this study provides false positive results and hence the test chemical is considered to be negative for gene mutation in vitro.