# Spelt Grass Juice: Phytochemicals and Antiproliferative Activity
## 3 Results and Discussion
### Antiproliferative Activity of Extracts
The apoptosis induced by the _Triticum aestivum_ extract was correlated with high free radical scavenging activity, supporting the role of antioxidants in reducing cancer cell viability [12]. Namely, as stated by these authors, an analysis of wheatgrass extract revealed the presence of biologically active compounds containing hydroxyl groups and double bonds, which help stabilize free radicals and, thus, exhibit antioxidant properties. Reactive oxygen species, or free radicals, are converted into hydrogen peroxide and an oxygen molecule by the antioxidant enzymes superoxide dismutase and cytochrome oxidase. This transformation leads to the destruction of cancer cells. Furthermore, [PERSON] et al. [2] demonstrated the antioxidant activity of spelt grass juice samples obtained from the same geographic region and produced by the same local manufacturer in Montenegro as the samples analyzed in our study. Their findings, based on FRAP, DPPH, and ABTS assays, revealed a positive correlation between antioxidant activity and the phenolic and flavonoid content of the juice. The antioxidant-dependent antiproliferative properties of _Triticum spelta_ could be significantly attributed to the presence of phenolic compounds and carotenoids, as well as their synergistic interactions [3; 28]. Numerous studies have highlighted the anticancer potential of ferulic acid, which is predominant in spelt grass juice extracts [29]. Phenolic acids, including \(p\)-coumaric acid, ferulic acid, and sinapic acid, have been shown to induce the dose-dependent inhibition of proliferation, concentration-dependent apoptosis, and HDAC inhibition in HeLa cells, with \(p\)-coumaric and ferulic acid specifically causing cell \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline \multicolumn{1}{c}{\multirow{2}{*}{**Spelt Grass Juice Extract**}} & \multicolumn{3}{c}{**IC50**} & \multicolumn{3}{c}{**NT/T**} \\ \cline{2-7}  & **MRC-5** & **A-549** & **HeLa** & **A-549** & **HeLa** \\ \hline Extract 1 & Aqueous (no foam) & 1319.30 & 1887.50 & 1162.10 & 0.70 & 1.14 \\ \hline Extract 2 & Aqueous (with foam) & 1405.90 & 2060.12 & 1184.97 & 0.68 & 1.19 \\ \hline Extract 3 & Ac:H\({}_{2}\)O (no foam) & 331.07 & 530.68 & 182.05 & 0.62 & 1.82 \\ \hline Extract 4 & Ac:H\({}_{2}\)O (with foam) & 416.84 & 529.20 & 203.07 & 0.79 & 2.05 \\ \hline Extract 5 & MeOH:H\({}_{2}\)O (no foam) & 271.96 & 453.22 & 221.82 & 0.60 & 1.23 \\ \hline Extract 6 & MeOH:H\({}_{2}\)O (with foam) & 293.13 & 411.13 & 228.89 & 0.71 & 1.28 \\ \hline \hline \end{tabular}
\end{table} 
Table 2: IC50 values for various spelt grass juice extract (\(\mathrm{\SIUnitSymbolMicro g}\)/mL) and their selectivity towards tumor cells (NT/T). cycle arrest at the S-phase [30]. Additionally, carotenoids, predominantly \(\alpha\)- and \(\beta\)-carotene, contribute to the strong antioxidant capacity of _Tritcium aestivum_ by scavenging free radicals and inhibiting reactive oxygen species (ROS), potentially enhancing its antiproliferative effects on cancer cells through synergistic interactions with other bioactive compounds [28]. Given the previously mentioned results, our research further confirms the relationship between the presence of antioxidant compounds and antiproliferative effects. The NT/T ratio indicates greater drug efficacy against tumor cells than toxicity to normal cells when exceeding 1.0 [31]. In our experiment, all six extracts showed selectivity toward HeLa cells, particularly acetone-water extracts, but not A-549 cells. The presence of photopigments such as chlorophyll and carotenoids (Figure 1) may contribute to this effect, as chlorophyll induces phase 2 detoxification enzymes and enhances oxygen delivery, which can be detrimental to oxygen-sensitive cancer cells [7]. Carotenoids, through their antioxidant properties, may further suppress tumor growth or trigger apoptosis [32].