Registration Dossier

Administrative data

Key value for chemical safety assessment

Additional information

Ames test:

In a bacterial reverse mutation test (Wild et al., 1983), alpha-Hexylcinnamaldehyde (HCA) was tested for toxicity similarly to OECD Guideline 471 (Bacterial Reverse Mutation Assay).

The strains S. typhimurium TA1535, TA1537, TA1538, TA98 and TA100 were treated with HCA at 5 concentrations up to 3.6 mg/plate (if not toxic, toxicity not reported) in DMSO, with and without activation with liver preparations (S9 mix) from rats treated with Aroclor 1254.This study was carried out using the standard plate incorporation method.Vehicle and positive controls were performed in both tests.

No biologically significant increase in the number of revertants was noted in any strain, either with or without metabolic activation.

The study does not meet modern standards where a limit of 5 mg/plate is required. The difference between 3.6 and 5.0 mg/plate is small and I it would be highly unusual to see something positive at 5.0 and negative at 3.6. Therefore the study is considered as acceptable.

Under the experimental conditions used and according to the positive criteria, it is concluded that the test substance was not mutagenic.

Mouse lymphoma assay:

Hexyl Cinnamic Aldehyde was tested for mutagenic potential in an in vitro mammalian cell mutation assay. This test system is based on detection and quantitation of forward mutation in the subline 3.7.2c of mouse lymphoma L5178Y cells, from the heterozygous condition at the thymidine kinase locus (TK+/-) to the thymidine kinase deficient genotype (TK-/-). The study consisted of a preliminary toxicity test and two main tests comprising three independent mutagenicity assays. The cells were exposed for either 3 hours or 24 hours in the absence of exogenous metabolic activation (S9 mix) or 3 hours in the presence of S9 mix.

No increase in mutant frequency exceeded the sum of the mean concurrent vehicle control mutant frequency. In all tests the concurrent vehicle and positive controls were within the acceptable historical control ranges.

It was concluded that Hexyl Cinnamic Aldehyde did not demonstrate mutagenic potential in this in vitro cell mutation assay, under the experimental conditions described.

Micro nucleus assay in vivo:

In an in vivo micronucleus assay (Wild et al., 1983), performed similarly to the OECD guideline No. 474 with minor deviations, male/female NMRI mice were exposed to alpha-Hexylcinnamaldehyde (HCA) diluted in olive oil by a single intraperitoneal dose. Different concentrations (324, 540, 756 mg/kg bw) were tested (8 animals per dose). 30 hours after the injection, the bone marrow cells were collected and one thousand polychromatic erythrocytes were observed in order to quantify the number of micronucleated polychromatic erythrocytes. Concurrent vehicle animals were used as negative control, no data was available on positive control.

Under the test conditions, there was no significant increase of micronucleated plychromatic erythrocytes, if compared with the concurrent control. No mortality was observed.

Therefore, under the study conditions it is concluded that HCA is neither clastogenic nor aneugenic.

In conclusion, the available data does not indicate potential genotoxicity. The endpoint on chromosomal aberration is covered by the in vivo micronucleus study. The data are considered to be adequate for risk assessment and classification and labelling.


Short description of key information:
In vitro bacterial reverse mutation assay (Ames test) was negative.
In vitro gene mutation assay using Mouse Lymphoma L5178Y cell based on OECD476 was negative.
In vivo micronucleus study using mice was negative.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Harmonized classification:

No harmonized classification is available according to the Regulation (EC) No 1272/2008 including ATP1.

Self-classification:

The available data are considered adequate for the purpose of classification and labelling. The evidence available does not justify the classification of Hexylcinnamaldehyde for genotoxicity. No self-classification is proposed according to the Directive 67/548/EEC and the Regulation (EC) No. 1272/2008 (CLP Regulation).