Pressurized liquid extraction followed by high-performance liquid chromatography for determination of beta-ecdysone...

Pfaffia glomerata (Spreng.) Pedersen has among its main bioactive compounds saponins, with the phytoestroid β-ecdysone as its chemical marker. In this study, pressurized liquid extraction (PLE), a green extraction technique used to obtain bioactive compounds from plants, was employed to extract beta-ecdysone from P. glomerata leaves, stems, and roots. The 2² factorial design was used with the variables temperature (333 K and 353 K) and flow rate (1.5 and 2 mL min^-1), pressure (300 Bar), time (60 min), and solvent [ethanol and distilled water (70:30 (v/v)] were kept constant for all parts of the plant. The results of experimental responses demonstrated that the factors temperature and flow rate significantly interfere with the yields of leaf (0.499%), root (0.65%) and stem (0.764%) extracts. The latter presented presents the highest yield compared to the other parts of the plant. HPLC results showed the presence of beta-ecdysone in all parts of the plant with concentrations of β-ecdysone 86.82, 76.53 and 195.86 mg L^-1 to leaf, root and stem, respectively. FT Raman results exhibited typical peaks of beta-ecdysone, such as 3310 cm^-1, 1654 cm^-1, and 1073 cm^-1 for all plant parts. Another interesting result was the presence of the peak at 1460 cm^-1 in the PLE root extract can be associated with selenium. This foundational knowledge confirms that the PLE extraction process was efficient in obtaining the chemical marker of Pfaffia glomerata in all plant parts.