## 2 Results and Discussion
### Cytotoxic and Hemolytic Effects of C. longa Extracts and Curcumin  
After 48 h of treatment, a dose-dependent effect was observed on cell death for both RAW 264.7 and VERO cell lines (Supplementary Material, Figure S4). The 50% cytotoxic concentration (CC\({}_{50}\)) values ranged from 70.16 to 47.8 \(\upmu\)g/mL for RAW 264.7 cells and from 89.3 to 58 \(\upmu\)g/mL for VERO cells (Table 3). These values were significantly higher than the CC\({}_{50}\) values of the reference drug control, PAT (potassium antimonyl(III) tartrate hydrate), as a source of trivalent antimony (Sb(III)), which were 30.8 \(\pm\) 0.73 \(\upmu\)g/mL for RAW 264.7 cells and 23.9 \(\pm\) 1.5 \(\upmu\)g/mL for VERO cells, respectively.  
\begin{table}
\begin{tabular}{l c c c c c c} \hline \hline \multirow{2}{*}{**Pharmacokinetic and Toxicological Endpoints**} & \multicolumn{6}{c}{**Compounds**} \\ \cline{2-7}  & **1** & **2** & **3** & **4** & **5** & **6** \\ \hline [PERSON]’s rule of 5 & 0 & 0 & 0 & 0 & 0 & 0 \\ Unbound (\%) to plasma protein & 6.7 & 6.2 & 7.7 & 8.9 & 12.1 & 11.2 \\ P-glycoprotein interaction & Yes & Yes & Yes & No & Yes & Yes \\ Mutagenicity (Ames test) & No & No & No & No & No & No \\ Carcinogenicity (rats) & No & No & No & No & No & No \\ Acute toxicity (rats) & No & No & No & No & No & No \\ Cardiotoxicity (hERG) & No & No & No & No & No & No \\ Hepatotoxicity & No & No & No & No & Yes & Yes \\ \hline Curcumin (**1**); demethylocurcumin (**2**); bsidemethoxycurcumin (**3**); ar-turmerone (**4**); \(\alpha\)-turmerone (**5**), and \(\beta\)-turmerone (**6**). & & & & \\ \end{tabular}
\end{table}
Table 2: In silico evaluation of pharmacokinetic and toxicological endpoints of the curcuminoids and turmerones.  
The curcuminoid-rich fraction showed approximately ten times higher cytotoxicity toward VERO cells compared to compound **3**, while the ethanolic extract was the least cytotoxic, with a CC\({}_{50}\) of 525 \(\mathrm{\SIUnitSymbolMicro}\)g/mL [46]. Scientific evidence indicates that curcumin (**1**), at non-toxic doses, exerts hepatotoxicity effects [47]. Moreover, **1** has demonstrated the potential to prevent and treat acute kidney injury induced by drugs such as amphoteric in B through a modulation of the immune response, a reduction of inflammatory markers, decreased apoptosis, and improved mitochondrial dynamics [48]. Clinical studies have also confirmed that the oral administration of standardized turmeric and curcumin extracts is safe for human use [49].  
Cytotoxicity was also evaluated through the hemolytic potential of _C. longa_ extracts against sheep red blood cells (RBC). The results demonstrated that there is a tendency for hemolysis to occur as the concentration of the extracts increases (Supplementary Material, Figure S4). [PERSON] and [PERSON] (2006) proposed a notable guide for classifying the hemolytic potential of pharmaceutical excipients, marking those with values exceeding 25% hemolysis as potentially hemolytic [50]. For _C. longa_ extracts, hemolysis did not exceed 25% up to 100 \(\mathrm{\SIUnitSymbolMicro}\)g/mL. The 50% hemolytic concentrations (CH\({}_{50}\)) were 119.65 \(\pm\) 3.4 (HEXCURC), 101.85 \(\pm\) 9.6 (DCCURC), and 144.15 \(\pm\) 13.35 (ETOHCURC) \(\mathrm{\SIUnitSymbolMicro}\)g/mL (Table 3). PAT did not show hemolytic potential at the highest concentration tested (100 \(\mathrm{\SIUnitSymbolMicro}\)g/mL).