Following a request from the European Commission, the EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids (CEF) was asked to deliver a scientific opinion on the implications for human health of chemically defined flavouring substances used in or on foodstuffs in the Member States. In particular, the Panel was asked to evaluate flavouring substances using the Procedure as referred to in the Commission Regulation (EC) No 1565/2000 0 (hereafter ‘the Procedure’).
The Union List of flavourings and source materials was established by Commission Implementing Regulation (EC) No 872/2012. The list contains flavouring substances for which the scientific evaluation should be completed in accordance with Commission Regulation (EC) No 1565/2000.
The Flavouring Group Evaluation 208 (FGE.208), corresponding to subgroup 2.2 of FGE.19, concerns three alicyclic aldehydes with the α,β-unsaturation in ring / side-chain and seven precursors for such. The α,β-unsaturated aldehyde structure, which is a structural alert for genotoxicity and the data on genotoxicity previously available for these 10 substances, did not rule out the concern for genotoxicity.
Among the 10 flavouring substances in FGE.19 subgroup 2.2, the Panel identified p-mentha-1,8-dien-7-al [FL-no: 05.117], for which appropriate genotoxicity data could be used for reading across to the other substances in the subgroup; therefore genotoxicity data have been requested for p-mentha-1,8-dien-7-al [FL-no: 05.117] according to the testing strategy worked out by the Panel.
In 2012, the Flavour Industry submitted new genotoxicity data: a bacterial reverse mutation assay, a hypoxanthine-guanine phosphoribosyl transferase (HPRT) assay and an in vitro micronucleus assay. These new data were evaluated in FGE.208 where the Panel concluded that the available data still gave rise to concern for the genotoxic potential of p-mentha-1,8-dien-7-al [FL-no: 05.117]. Therefore, the Panel asked to provide an in vivo Comet assay performed on the first site of contact (e.g. stomach or duodenum) and on liver.
The Flavour Industry has submitted a combined study in rats: a bone marrow micronucleus test and Comet assay in liver and duodenum. This study is evaluated in the present Revision 1 of FGE.208 (FGE.208Rev1). p-Mentha-1,8-dien-7-al [FL-no: 05.117] was administered to rats by oral gavage at three dose levels: 175, 350 and 700 mg/kg bw/day (which is an estimate of the maximum tolerated dose). The results of the micronucleus assay suggest that p-mentha-1,8-dien-7-al [FL-no: 05.117] did not induce any increase in micronucleated polychromatic erythrocytes.
In the same animals, results of the Comet assay suggest that p-mentha-1,8-dien-7-al [FL-no: 05.117] did not induce any DNA damage in duodenum, but a statistically significant increase in DNA strand breaks was observed in the liver at the highest dose tested (700 mg/kg bw/day).
The Panel noted that the results observed at the highest dose were more than 3-fold higher than the concurrent negative control value and statistically significant different from the negative control value, and a statistically significant positive linear trend was observed. The Panel considered that since there was a wide range of historical control data with an overlap of the positive and negative historical control values, the historical control data could not be used as a criterion to interpret the data.
Overall, the Panel concluded that p-mentha-1,8-dien-7-al [FL-no: 05.117] is genotoxic in vivo and that, accordingly, there is a safety concern for its use as flavouring substance.
Since p-mentha-1,8-dien-7-al [FL-no: 05.117] is representative for the nine remaining substances of this subgroup 2.2 (p-mentha-1,8-dien-7-ol [FL-no: 02.060], myrtenol [FL-no: 02.091], myrtenal [FL-no: 05.106], 2,6,6-trimethyl-1-cyclohexen-1-carboxaldehyde [FL-no: 05.121], myrtenyl formate [FL-no: 09.272], p-mentha-1,8-dien-7-yl acetate [FL-no: 09.278], myrtenyl acetate [FL-no: 09.302], myrtenyl-2-methylbutyrate [FL-no: 09.899] and myrtenyl-3-methylbutyrate [FL-no: 09.900]), there is a potential safety concern for these substances.