## 4 Conclusions  
Throughout history, _Iberis_ species have been researched for the presence of GSLs using various approaches. These studies revealed that they primarily contain GSLs derived from \begin{table}
\begin{tabular}{c c c c c c} \hline \hline \multirow{2}{*}{**Cell Line**} & \multicolumn{4}{c}{**IC50 (\(\mu\)g/mL)**} & \multirow{2}{*}{**Control a**} \\ \cline{2-2} \cline{4-5}  & **HD** & **MAD** & **EXT** & **MHG** & \\ \hline A549 & 8.45 & 9.42 & 1.46 & 36.41 & 84.71\% \\ T24 & 0.61 & 0.62 & 1.83 & 23.30 & 52.32\% \\ \hline \hline \end{tabular} HD—hydrodistillate; MAD—microwave-assisted distillate; EXT—CH\({}_{2}\)Cl\({}_{2}\) extract after the 24 h autolysis with added myrosinase; MHG—extract obtained by microwave hydrodiffusion and gravity. \({}^{*}\) The percentages of metabolically active cells (as % total cells) for cisplatin as a positive control (50 ug/mL) after 72 h.  
\end{table} 
Table 4: Calculated IC50 values (ug/mL) for volatiles obtained by HD, MAD, EXT, and MHG from the whole plant material (seed, stem, leaf, and flower) against human lung cancer cell A549 and bladder cancer cell T24 lines after 72 h.  
Figure 3: Percentage of metabolically active cells in human lung cell line A549 and human bladder cell line T24 after 72 h of incubation with different concentrations of _I. sempervirens_ samples. HD—hydrodistillate; MAD—microwave-assisted distillate; EXT—CH\({}_{2}\)Cl\({}_{2}\) extract after the 24 h autolysis with added myrosinase; MHG—extract obtained by microwave hydrodiffusion and gravity. Calculated IC50 values (ug/mL) are given in Table 4. Each data point is presented as the mean \(\pm\) SD (n = 3). Lowercase letters represent the significance level in comparison to non-treated cell line samples (a, \(p\) < 0.001; b, \(p\) < 0.01; c, \(p\) < 0.05). methionine. This was supported by phylogenetic studies that link the evolution of both the Brassicaceae family and the related GSLs. However, some of the research on these plant species was carried out by utilizing indirect methods of GSL identification, i.e., GC-MS analyses of their breakdown products. This and other studies suggest that some former findings should be re-evaluated using suitable spectroscopic techniques for direct GSL identification. GSL breakdown products, usually ITCs, are mostly associated with the observed bioactive properties, as opposed to the other volatiles that can be formed, such as nitriles. This work underlines the significance of adopting appropriate conditions for GSL degradation (thermal or enzymatic) as well as the selection of the method used for isolation, such as distillation or extraction, due to the differences in polarity and molecular weight. It is also evident that the oxidation state of the sulfur in the side chain, next to the ITC functional group, influences the bioactivity potential as well. The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/app14010346/s1](https://www.mdpi.com/article/10.3390/app14010346/s1)]([https://www.mdpi.com/article/10.3390/app14010346/s1](https://www.mdpi.com/article/10.3390/app14010346/s1)), Figure S1: Chromatograms of desulfoglucosionolates (dGSLs) obtained from the different plant parts of _I. semperviens._**d73**--3-(methylsulfonyl)propyl dGSL (desulfoglucoiberini); **d95**--3-(methylsulfonyl)propyl dGSL (desulfoglucoiberini); **d84**--4-(methylsulfonyl)butyl dGSL (desulfoglucoerini); **d11**--benzyl dGSL (desulfoglucotropaeolin); **d48**--4-4-methoxyindol-3-ylmethyl dGSL (desulfo-4-methoxyglucobrassicin); Figure S2: MS\({}^{2}\) spectra of desulfoglucosionates from _I. semperviens._**d73**--3-(methylsulfonyl)propyl dGSL (desulfoglucoiberini); **d95**--3-(methylsulfonyl)propyl dGSL (desulfoglucoiberini); **d84**--4-(methylsulfonyl)butyl dGSL (desulfoglucoerucin); **d11**--benzyl dGSL (desulfoglucotropaeolin); **d48**--4-methoxyindol-3-ylmethyl dGSL (desulfo-4-methoxyglucobrassicin). Chemistry investigation: A.D., F.B. and I.B. ; antiproliferative assays: V.C.C. ; writing--original draft: A.D. and I.B.