Determination of Progesterone Compounds in the Crude Methanol Extract of Benalu Duku Leaves

Background And Aim: L. Miq (benalu duku) is a parasitic herb that commonly grows on the host plant . Researchers have found that the plant contains anticancer compounds and may contain phytoandrogens, including progesterone-like compounds, in its crude methanol extract. The objective of the current study was to investigate the compound of phyto progesterone in benalu duku leaves after extracted by methanol and prepared using an analytical column of high-performance liquid chromatography (HPLC).

Materials And Methods: About 400 g of benalu duku leaves were pulverized, and their compounds were isolated by the isocratic method using an RP-18 analytical column (5 µm) with a mobile phase of 70:30 (methanol: water) in a photodiode array detector adjusted to 254 nm. The phyto progesterone compound was identified at a retention time of approximately 6.01 min.

Results: By LC-electrospray ionization mass spectrometry focusing on molecular fractions, the fingerprint area of the Fourier transform-infrared spectroscopy (FT-IR, cm) and nuclear magnetic resonance (NMR) spectra indicated that the phyto progesterone product isolated was identical to the certified reference material of pure progesterone, particularly the specific functional groups in the FT-IR spectrum at wavenumbers of 1317.43 cm and 1386.86 cm and in the proton NMR spectrum at carbon 21 of progesterone (p<0.05).

Conclusion: Each 49.888 µg/mL of crude benalu duku leaf extract dissolved in the mobile phase contained 28.515±0.713 µg/mL phyto progesterone.