## 3 Results and Discussion
### Biological Activities of Sea Fennel Leaf, Flower, and Fruit EOs and Hydrolates
Figure 2: Percentage of metabolically active human breast adenocarcinoma MDA-MB-231, human breast metastatic adenocarcinoma MCF-7, and human ovarian carcinoma OVCAR-3 cell lines after 4, 24, 48, and 72 h of incubation with different concentrations of sea fennel essential oils and hydrolates from leaves, flowers, and fruits. Since essential oils and hydrolates contain a wide range of diverse components, and their cytotoxic activity was tested at different concentrations and exposure times, a linear regression analysis was performed to evaluate the impact of various factors on the cytotoxic activity of EOs and hydrolates from sea fennel leaves, flowers, and fruits (Tables 4 and 5). The concentration of the EOs had the greatest effect on the MCF-7 cell line (B = \(-\)0.226, \(p\) < 0.0001), with a lower IC\({}_{50}\) value observed at higher concentrations. The concentration was a significant factor for all the cell lines tested. For the MCF-7 and MDA-MB-232 cell lines, octanal showed a significant effect (B = 75.86, \(p\) < 0.0001 and B = 74.10, \(p\) < 0.0001, respectively). The only factor that had a significant effect on all the hydrolate-treated cell lines was time (Table 5). For all three cell lines (MCF-7, MDA-MB-232, and OVCAR-3), the coefficients were negative (B = \(-\)0.28, B = \(-\)1.51, and B = \(-\)0.254, respectively), indicating that the IC\({}_{50}\) values decreased with increasing exposure time. Regarding the chemistry of essential oils, it can be suggested that the high concentration of limonene plays a key role in the strong cytotoxic activity of sea fennel EOs. Limonene is well known for its chemopre properties against various types of cancer and possesses no mutagenic, carcinogenic, or nephrotoxic risks to humans [50]. However, the presence of \(\alpha\)-pinene [51] and several other biologically active compounds suggests that the cytotoxic effects observed for sea fennel EOs may be due to their synergistic effect. Since hydrolates are highly diluted, they generally exhibit less biological activity than their corresponding essential oils. However, due to the presence of a wide variety of compounds with anticancer properties, such as teripinen-4-ol, \(\alpha\)-terpineol, carvone, and carveol [38; 39; 40], as well as their probable synergistic effects, the sea fennel hydrolates also demonstrated cytotoxic activity. \begin{table}
\begin{tabular}{c c c c c c c} \hline
**Cell Line** & **Predictor** & **Coefficient** & **Standard Error** & **F-Value** & **R\({}^{2}\)** & _p-Value_ \\ \hline \multirow{4}{*}{**MCF-7**} & Concentration (\(\mu\)g/mL) & \(-\)0.226 & 0.03 & & & \(<\)0.0001 \\  & Octanal & 75.86 & 13.01 & & & \(<\)0.0001 \\  & Intercept & 91.10 & 5.153 & 48.00 & 0.627 & \(<\)0.0001 \\ \hline \multirow{4}{*}{**MDA-MB-232**} & Concentration (\(\mu\)g/mL) & \(-\)0.17 & 0.03 & & & \(<\)0.0001 \\  & Octanal & 74.10 & 11.81 & & & \(<\)0.0001 \\  & Time (h) & \(-\)0.43 & 0.09 & & & \(<\)0.0001 \\  & Intercept & 110.66 & 5.83 & 33.00 & 0.645 & \(<\)0.0001 \\ \hline \multirow{4}{*}{**OVCAR-3**} & Concentration (\(\mu\)g/mL) & \(-\)0.064 & 0.016 & & & \(<\)0.0001 \\  & Dodecane & 36.695 & 7.174 & & & \(<\)0.0001 \\ \cline{1-1}  & Intercept & 85.833 & 3.018 & 21.56 & 0.481 & \(<\)0.0001 \\ \hline \end{tabular}
\end{table} 
Table 4: Results of linear regression for the cytotoxicity of sea fennel leaf, flower, and fruit essential oils on cancer (MDA-MB-231, MCF-7, and OVCAR-3) cell lines. \begin{table}
\begin{tabular}{c c c c c c} \hline
**Cell Line** & **Predictor** & **Coefficient** & **Standard Error** & **F-Value** & **R\({}^{2}\)** & _p-Value_ \\ \hline \multirow{2}{*}{**MCF-7**} & Time (h) & \(-\)0.28 & 0.11 & & & 0.025 \\  & Intercept & 96.48 & 4.853 & 6.92 & 0.409 & \(<\)0.0001 \\ \hline \multirow{2}{*}{**MDA-MB-232**} & Time (h) & \(-\)1.51 & 0.17 & & & \(<\)0.0001 \\  & Intercept & 137.88 & 7.54 & 80.93 & 0.89 & \(<\)0.0001 \\ \hline \multirow{2}{*}{**OVCAR-3**} & Time (h) & \(-\)0.254 & 0.085 & & & 0.014 \\  & Intercept & 77.477 & 3.835 & 8.83 & 0.47 & \(<\)0.0001 \\ \hline \end{tabular}
\end{table}