# Spelt Grass Juice: Phytochemicals and Antiproliferative Activity
## 3 Results and Discussion
### Antiproliferative Activity of Extracts  
The antiproliferative effects of six whole extracts derived from spelt grass juice were evaluated on three distinct cell lines. This study aimed to assess the influence of the extract type and solvent choice on their antiproliferative properties as well as the selectivity of the extracts towards two malignant cell lines, and their potential to enhance the efficacy of doxorubicin in co-treatment. All tested spelt grass juice extracts, irrespective of the presence of foam, exhibited a concentration-dependent antiproliferative effect (Figure 2). Co-treatment with doxorubicin led to an enhanced inhibitory response across all examined cell lines. The presence of foam did not induce significant changes in the observed effects. Figure 2: Conf. Figure 2: Antiproliferative effect of spell grass juice extracts on cell growth after 48 h incubation at different concentrations, individually and in combination with 2 μm downchrobin. Cell lines tested: (a) MRC\(\,\leq\,\)–onto-transformed cells (b) A-569 lung cancer cells; and (d) HeLa–cruver cancer cells. Extracts Extract 1 – Aqueous extract of spell grass juice without foam. Extract 2 –Auqueous extract of spell grass juice with foam; Extract 3 –AcH\(\,\)O (14) extract of lyophilized spell grass juice without foam; Extract 4 –AcH\(\,\)O (41) extract of lyophilized spell grass juice without foam; Extract 6 –MoCHH\(\,\)O (41) extract of lyophilized spell grass juice with foam; Extract 5 –MoCHH\(\,\)O (41) extract of lyophilized spell grass juice without foam; and Extract 6 –MoCHH\(\,\)O (41) extract of lyophilized spell grass juice without foam, and their combination with Dox (downchrobin). Concentrations tested: Conc. 1–233 \(\mu\)g/mL (Extract 1), 6633 \(\mu\)g/mL (Extract 2), and 100 \(\mu\)g/mL (Extracts 3–6); Conc. 2–1290 \(\mu\)g/mL (Extract 2), and 300 \(\mu\)g/mL (Extract 3–6); Conc. 3–3200 \(\mu\)g/mL) (Extract 1, 3350 \(\mu\)g/mL (Extract 2), and 500 \(\mu\)g/mL (Extract 3–6); and Conc. 4–6400 \(\mu\)g/mL (Extract 1), 6700 \(\mu\)g/mL (Extract 2), and 1000 \(\mu\)g/mL (Extracts 3–6). All results are expressed by end of growth relative to the untreated control groups. Statistically significant differences between means at each concentration, indicated by different letters above bars, were determined using one-way ANOVA and Duncan’s test at \(p<0.05\). Solvent selection, determined by variations in polarity and solubility, significantly influenced the extraction efficiency and the phytochemical composition of the obtained extracts. Among the tested extracts, the methanol-water fraction demonstrated the most potent inhibitory effect on MRC-5 normal fibroblasts and A-549 lung cancer cells, whereas the acetone-water extract exhibited the highest antiproliferative activity against HeLa cervical carcinoma cells, likely due to its ability to extract both polar and non-polar bioactive compound. In contrast, the aqueous extract consistently showed the weakest inhibitory effects across all tested cell lines (Table 2). These findings underscore the importance of solvent selection in the extraction process, as it significantly influences the phytochemical composition and, consequently, the biological activity of the extracts. Regarding the IC50 values for all types of extracts, cervical cancer cells were more susceptible compared to lung cancer cells. The higher efficacy towards cervical cancer cells was observed for the acetone-water and methanol-water extracts compared to aqueous ones. Our results align with previous studies [5; 17] that demonstrated the antiproliferative properties of _Triticum aestivum_ extracts on the HeLa cell line. [PERSON] et al. [5] showed that the methanolic leaf extract of _Triticum aestivum_ exhibited moderate anticancer activity with an IC50 value of 156 \(\mathrm{\SIUnitSymbolMicro g}\)/mL against HeLa cervical carcinoma cells.