# Phenolic Compound Composition and Biological Activities of Fractionated Soybean Pod Extract
## 3 Results and Discussion
The fractions with variable lipophilicity isolated from the ethanol extract were tested for their cytotoxic activity against human colorectal carcinoma (HCT116) and prostate adenocarcinoma (PC-3) cells. The L929 cell line, i.e., mouse fibroblasts from subcutaneous adipose tissue, was used as a normal cell model, as recommended for research associated with the cytotoxicity of chemical compounds and the comparison of other results. The cytotoxic effects of the investigated samples depended on the type of cancer cells, and did not exceed 160 \(\mathrm{\SIUnitSymbolMicro g}\)/mL (Table 1). It was shown that the PC-3 cell line was the most sensitive to the effect of the soybean pod extracts and fractions with variable lipophilicity. \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline \multicolumn{2}{c}{**Total Phenolic Content 1**} & \multicolumn{2}{c}{**Antioxidant Activity 2**} & \multicolumn{2}{c}{**Cytotoxic Activity 3**} & \\ \hline  & DPPH & ABTS & HCT116 & PC-3 & I929 \\ \hline E & 24.649  \(\pm\) 0.618 & 427.3  \(\pm\) 4.1 & 73.1  \(\pm\) 1.1 & 116  \(\pm\) 3.7 & 54  \(\pm\) 3.1 & 70  \(\pm\) 2.6 \\ F1 & 0.016  \(\pm\) 0.019 & 2668.7  \(\pm\) 10.3 & 956.5  \(\pm\) 8.3 & 102  \(\pm\) 3.4 & 56  \(\pm\) 2.1 & 56  \(\pm\) 2.1 & 56  \(\pm\) 2.1 & 56  \(\pm\) 2.1 & 56  \(\pm\) 2.1 & 56  \(\pm\) 2.1 & 14.510  \(\pm\) 0.097 & 320.2  \(\pm\) 3.5 & 154.7  \(\pm\) 2.4 & 156  \(\pm\) 3.3 & 66  \(\pm\) 2.8 & 112  \(\pm\) 2.8 & 112  \(\pm\) 2.8 \\ F3 & 6.028  \(\pm\) 0.214 & 510.6  \(\pm\) 5.2 & 320.2  \(\pm\) 4.7 & 120  \(\pm\) 3.1 & 70  \(\pm\) 2.9 & 80  \(\pm\) 3.3 \\
5-fluorouracil 4 & n.a  & n.a  & 33 [FOOT:3]Footnote 3: footnotemark: [ENDFOOTNOTE] \(\pm\) 3.2 & 23  \(\pm\) 2.58 & 8  \(\pm\) 1.1 \\ \hline \end{tabular}
\end{table} 
Table 2: Total phenolic content and antioxidant and cytotoxic activity of 80% ethanolic extract (E), water (F1), 40% methanol–water (F2), and 70% methanol (F3) fractions obtained from soybean pods. \begin{table}
\begin{tabular}{c c c c} \hline \hline  & **DPPH** & **ABTS** & **TPC** \\ \hline TPC & \(-\)0.99188 & \(-\)0.99941 & - \\ Cytoxic activity (PC-3) & 0.97829 & 0.949829 & \(-\)0.94401 \\ \hline \hline \end{tabular}
\end{table} 
Table 3: [PERSON]’s correlation coefficient. A significantly lower degree of cytotoxicity was noted in the experimental series with the use of normal cells from cell line L929. The F2 fraction with medium lipophilicity produced the most pronounced cytotoxic effect in the PC-3 cells (IC\({}_{50}\) = 66 \(\upmu\)g/mL), which was two times higher than in the case of the L929 cells (IC\({}_{50}\) = 112 \(\upmu\)g/mL); the 80% ethanol extract was very effective as well (IC\({}_{50}\) = 54 \(\upmu\)g/mL). The aqueous fraction (F1) did not exhibit PC3-specific cytotoxicity (IC\({}_{50}\) = 56 \(\upmu\)g/mL for PC-3 and L929). The analysis of [PERSON]'s correlation (Table 3) demonstrated a high relationship between the content of phenolic compounds and the values of IC\({}_{50}\) which reflects the antioxidant activity of the extracts and fractions and the cytotoxic activity towards the PC-3 cells. The colorectal carcinoma cells showed slight sensitivity to the action of the pod extract and the fraction with variable lipophilicity. This is illustrated by the IC\({}_{50}\) values from 102 to 156 \(\upmu\)g/mL, which markedly exceed the values obtained for L929 cells. A higher degree of cytotoxicity was noted in the experimental series using normal cells (IC\({}_{50}\) from 56 to 112 \(\upmu\)g/mL). One of the preliminary tests necessary to determine the proper action and safety of a potential drug are cytotoxicity studies using in vitro cell cultures. Although the conventional in vitro methods used to determine cytotoxicity have lower specificity or ability to evaluate the mechanism of action than animal models, they are a widely used and useful tool for screening new compounds. In the case of anti-tumor activity, limited information is available in the literature on soybean pod extract. [PERSON] et al.