2-benzylideneheptanal

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The mutagenicity effect of α-Amylcinnamaldehyde was studied by performing experiment in Salmonella typhimurium. Salmonella typhimurium stains TA1535, TA100, TA1537, TA1538, TA98 were used in study. Mutagenic assay was performed by plate incorporation method with and without metabolic activation i.e. S9 mix prepared from liver fraction of Aroclor pretreated rats (Aroclor 1254). DMSO used as solvent for test substance. Five doses of each substance were tested (up to 3.6 mg/plate) in all five tester strains Vogel Bonnet medium was used throughout; plates were incubated for 48 hr. Positive controls were run in each experiment with the reference mutagens sodium azide and benzo[a]pyrene. Test substance was tested at least twice.Test substance did not induce mutation in the tester strains, therefore α-Amylcinnamaldehyde was estimated to be not mutagenic in Salmonella typhimurium strains.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

No data

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:

Peer reviewed journal research article

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Deviations:

not specified

Principles of method if other than guideline:

The mutagenicity potential of α-Amylcinnamaldehyde by conducting experiment in Salmonella typhimurium TA1535, TA100, TA1537, TA1538 and TA 98.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

- Name of test material (as cited in study report): α-Amylcinnamaldehyde
- Molecular formula (if other than submission substance): C14H18O
- Molecular weight (if other than submission substance): 202.2952 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Target gene:

Histidine

Species / strain / cell type:

other: Species: Salmonella typhimurium Strain: TA1535, TA100, TA1537, TA1538, TA98

Details on mammalian cell type (if applicable):

Not applicable.

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mixture from liver fraction of rat pretreated with Aroclor 1254

Test concentrations with justification for top dose:

Up to 3.6 mg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Solubility of test substance in DMSO solvent.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: Plate incorporation

DURATION
- Preincubation period: No data
- Exposure duration: 48 hrs
- Expression time (cells in growth medium): No data
- 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: Two

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

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

Rationale for test conditions:

No data

Evaluation criteria:

Dose dependent increase in number of revertants.

Statistics:

Kastenbaum and Bowman Method

Species / strain:

other: Species: Salmonella typhimurium Strain: TA1535, TA100, TA1537, TA1538, TA98

Metabolic activation:

with and without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not valid

Positive controls validity:

valid

Additional information on results:

No data

Remarks on result:

other: No mutagenic effect were observed

Conclusions:

The mutagenicity potential of α-Amylcinnamaldehyde was evaluated by performing mutagenicity assay in Salmonella typhimurium strains TA1535, TA100, TA1537, TA1538, TA98, test substance did not induce mutation in all bacterial tester strains. Therefore α-Amylcinnamaldehyde was considered to be not mutagenic in Salmonella typhimurium strains.

Executive summary:

The mutagenicity effect of α-Amylcinnamaldehyde was studied by performing experiment in Salmonella typhimurium. Salmonella typhimurium stains TA1535, TA100, TA1537, TA1538, TA98 were used in study. Mutagenic assay was performed by plate incorporation method with and without metabolic activation i.e. S9 mix prepared from liver fraction of Aroclor pretreated rats (Aroclor 1254). DMSO used as solvent for test substance. Five doses of each substance were tested (up to 3.6 mg/plate) in all five tester strains Vogel Bonnet medium was used throughout; plates were incubated for 48 hr. Positive controls were run in each experiment with the reference mutagens sodium azide and benzo[a]pyrene. Test substance was tested at least twice.

Test substance did not induce mutation in the tester strains, therefore α-Amylcinnamaldehyde was considered to be not mutagenic in Salmonella typhimurium strains.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Based on the various experimental data for the target chemical have been reviewed to determine the mutagenic nature of alpha-Amyl cinnamaldehyde. The studies are as mentioned below:

  Based on the experimental key study conducted by Wild, et.al. (Food and chemical toxicology, 1983) .The mutagenicity effect of α-Amylcinnamaldehyde was studied by performing experiment in Salmonella typhimurium stains TA1535, TA100, TA1537, TA1538, TA98. Mutagenic assay was performed by plate incorporation method with and without metabolic activation i.e. S9 mix prepared from liver fraction of Aroclor pretreated rats (Aroclor 1254). DMSO used as solvent for test substance. Five doses of each substance were tested (up to 3.6 mg/plate) in all five tester strains Vogel Bonnet medium was used throughout; plates were incubated for 48 hr. Positive controls were run in each experiment with the reference mutagens sodium azide and benzo[a]pyrene. Test substance was tested at least twice. Test substance did not induce mutation in the tester strains, therefore α-Amylcinnamaldehyde was considered to be not mutagenic in Salmonella typhimurium strains.

  Similarly in the another study, where data is from authoritative database HSDB (2017) were observed . In vitro genetic toxicity study to Salmonella typhimurium species and strainsTA 1535, TA 100, TA 1537, TA 1538, TA 98 by using alpha-Amyl cinnamaldehyde in the presence of exogenous metabolic activation was prepared from Aroclor-pretreated rats (Aroclor 1254, 500 mg/kg).Toxicity checked at the maximum concentration. The test substance was exposed at the concentration of 3600 ug/plate .alpha-Amyl cinnamaldehyde did not induce a toxicity, dose-related increase in his+revertants over the corresponding solvent in theS. typhimurium tester strains TA 1535, TA 100, TA 1537, TA 1538, TA 98 in the presence of S9 metabolic activation system and hence is negative for mutation in vitro. Hence the test substance can not be classified as gene mutant in vitro. 

Similarly another experimental study was performed were data is from handbook and collection databases (HSDB, 2016 and HPVIS, 2017).  In vitro genetic toxicity study toSalmonella typhimurium species and strainsTA1535, TA100, TA1537, TA98 by using alpha-Amyl cinnamaldehyde in the absence and presence of exogenous metabolic activation was prepared from Aroclor-pretreated rats (Aroclor 1254, 500 mg/kg).Toxicity checked at the maximum concentration. Solvent control, DMSO was also run simultaneously.Based on the dose concentration 2.5 to 750 µg/plate without activation and 250 µg/plate with activation, alpha-Amyl cinnamaldehydedid not induce a toxicity, dose-related increase in his+revertants over the corresponding solventin theS. typhimurium tester strains TA1535, TA100, TA1537, TA98 in the presence of S9 metabolic activation system and in the absence .Hence is negative for mutation in vitro.

Similarly another studies was performed were data is from handbook and collection data (HSDB, 2016 and HPVIS, 2017). In vitro genetic toxicity study on Salmonella typhimurium species and strainsTA97 and TA102 by using alpha-Amyl cinnamaldehyde in the absence and presence of exogenous metabolic activation was prepared from Aroclor-pretreated rats (Aroclor 1254, 500 mg/kg).Toxicity checked at the maximum concentration. 9-aminoacridine was taken as a positive control for TA97 (with and without) and TA102 without activation, positive control for TA102 without S9 was mitomycin C.Based on the above concentration 1-1000 µg/plate alpha-Amyl cinnamaldehyde did not induce a toxicity, dose-related increase in his+revertants over the corresponding solvent in the S. typhimurium tester strains TA97 and TA102 in the presence of S9 metabolic activation system and absence hence is negative for mutation in vitro. 

Similarly study was conducted by Hiroshi Fujita and Mieko Sasaki(Annual Report of Tokyo Metropolitan Research Laboratory of Public Health, 1987). The Mutagenic potential of α-Amyl Cinnamaldehyde was evaluated by conducting experiment in Salmonella typhimurium StrainsTA97, TA102 with and without metabolic activation i.e. S9 mix. Mutagenic assay was performed by Preincubation method describe by Ames.DMSO was used as vehicle for test substance. Test chemical in doses0, 0.001, 0.005, 0.01, 0.5, 1 mg/plate were studied in mutagenic assay.Test chemical was preincubated with S9 mix for 20 min. Positive control and vehicle control also run parallel to test chemical.  The number of revertants in test chemical treated plates were less than solvent control as dose dependent increase in number of revertants not reported hence test chemical did not induce mutation. Therefore alpha -Amyl Cinnamaldehyde was not mutagenic in Salmonella typhimurium. 

Based on the data available for the target chemical α-Amyl Cinnamaldehyde does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the target chemical α-Amyl Cinnamaldehyde does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro as per the criteria mentioned in CLP regulation.