## 3 Results
### Food Samples, Choice, and Preparation  
Several samples of model food and drink preparations with SO were prepared. The effects of bioactive volatiles ITCs are thought to occur mainly locally in the upper airways and oropharyngeal cavity, i.e., in the nose, mouth, throat, larynx, and vocal cords [7]. Here, the pungency of ITCs, due to the interaction with TRPA1 ion channel, is perceived. For food preparations, we focussed on those expected to maximise the release of ITCs during/after the ingestion and possibly having a good taste at the same time.  
The first sample we prepared is honey added with fresh erysimum flowers or seeds and allowed to stand for 4 weeks.  
Honey is often used as an emollient for throat pain and mild affections of respiratory tract; honey with SO flowers (samples S2 and S3) or seeds (samples S4 and S5) can be stored for a long time at room temperature, preserving plant parts from spoiling; it can be consumed as such in a spoon or dissolved in hot water to make a sweet hot drink. The effect on the release of ITCS by leaving whole material or by grinding seeds or flowers during the heating, in order to simulate the mastication, was also studied.  
An infusion (S6) was prepared with commercial dry plant (whole aerial parts) in infusion in hot water, as suggested for standard herbal tea. As an alternative, a model voice drink (S7) was prepared with fresh seeds and flowers in water.  
Two samples were prepared to explore the effects of fermentation on SO active ITCs. Artisanal beer (S8) (\"the singer's beer\") was prepared by adding commercial dry SO in infusion during the malt fermentation process. A fermented tea with SO (S9) was prepared. Fermented drinks in the Kombucha style, also known as \"bubble teas\", are becoming popular among consumers for their pleasant sour taste, light carbonation, low alcohol content, and benefits on health [27].  
As a reference for commercial products using erysimum as an herbal remedy for voice, we analysed in the same experimental conditions a commercial tablet with SO extracts commonly used for aphonia (S10). All food and drink samples are listed in Table 2.  
\begin{table}
\begin{tabular}{c c c c c c} \hline \hline \multirow{2}{*}{**Entry**} & \multirow{2}{*}{**Samples 1**} & **2** & **4** & **Total ITCs** & **Total ITCs** \\  & & **(\(\upg/\)kg)** & (\(\upg/\)kg) & (\(\upg/\)kg) & **Portion** & **(\(\upg/\)**)** \\ \hline S2 & Honey FW & 3.10 \(\pm\) 0.63 & 0.25 \(\pm\) 0.01 & 3.51 & 7 g & 0.02 \\ S3 & Honey FG & 6.11 \(\pm\) 0.65 & 0.70 \(\pm\) 0.09 & 6.71 & 7 g & 0.05 \\ S4 & Honey SW & 3.98 \(\pm\) 1.13 & 0.30 \(\pm\) 0.18 & 4.01 & 7 g & 0.03 \\ S5 & Honey SG & 204.06 \(\pm\) 17.90 & 18.28 \(\pm\) 3.50 & 222.34 & 7 g & 1.55 \\ S6 & Infusion 2 & 38.56 \(\pm\) 8.51 & 8.29 \(\pm\) 1.00 & 46.84 & 125 mL & 5.85 \\ S7 & Voice drink & 4.55 \(\pm\) 1.13 & 3.06 \(\pm\) 1.60 & 7.61 & 125 mL & 0.95 \\ S8 & Beer & 19.64 \(\pm\) 0.44 & 3.46 \(\pm\) 0.76 & 23.11 & 330 mL & 7.62 \\ S9 & Kombucha & nd & nd & - & - & - \\ S10 & Tablet 3 & 1.71 \(\pm\) 0.11 & 0.20 \(\pm\) 0.02 & 1.91 & 1 (24 g) & 0.04 \\ \hline \hline \end{tabular} 1
[FOOTNOTE:1]Footnote 1: footnotetext: Legenda: F = flowers; S = seeds; W = whole; G = grinded. 2  
\end{table}
Table 2: Analysis of IPITC (2) and SBITC (4) released in food and drink preparations. Total ITCs (2 + 4) are reported as \(\upg/\)Kg of preparation.