Determination of progesterone compounds in the crude methanol extract of benalu duku leaves

Abstract

Background and Aim: Dendrophthoe pentandra L. Miq (benalu duku) is a parasitic herb that commonly grows on the host plant Lansium domesticum. 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−1) and Hnuclear 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−1 and 1386.86 cm−1 and in the proton HNMR 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.

In Vivo Analysis of Extract of Leaves of Mistletoe as a Benalu Duku: Clinical Chemical Value Associated with Histopathology, Liver, Kidneys, and Lethal Dose Determinate

“…HPLC semipreparation UV-photodiode array (PDA) detection was performed using Shimadzu LC-6AD pump with a DGU-20A5 degasser, communication module-20 A, and PDA detector SPD-M20A with an FRC-10A fraction collector. The UV detector for HPLC was set at 254 nm with a reverse-phase ODS C 18 column for analysis or semipreparation column Zorbax Eclipse XDB-C 18 at length of 9.4 × 250 mm, 5 μ m. The mobile phase was methanol: water at a ratio of 70 : 30 [ 5 ].…”

“…Then, approximately 20 g of crude extract was vacuum-dried and evaporated with nitrogen gas at 40°C. The crude extract was prepared to obtain steroid compounds and then the following purity by semipreparation of HPLC for obtaining PLCs [ 5 ]. The PLCs were identified and characterized using thermogravimetric analysis, FT-IR spectroscopy, and NMR by comparing them to the CRM.…”

“…The sample size was computed using the ( 1 ) below with the following: assumed { Z 1 − ( α /2)} at 1.96, a significance of 0.05; Z β at 1.645 by error limit of 5%; d at 3.62; S a at 1.7, and S b at 1.4 [ 5 ]. The N value was rounded to 5 rats for each treatment dose; thus, the control group should consists of five sets of data.…”

“…Te UV detector for HPLC was set at 254 nm with a reverse-phase ODS C 18 column for analysis or semipreparation column Zorbax Eclipse XDB-C 18 at length of 9.4 × 250 mm, 5 μm. Te mobile phase was methanol: water at a ratio of 70 : 30 [5].…”

“…Te host plant is well distributed in many countries worldwide and is known to grow well in tropical and subtropical countries [3,4]. Crude methanol extract of BD leaves contained 7.622% (w/w) essential amino acids from 4.384 mg of pulverized leaves before extraction [5]. Several reports between 2009 and 2010 showed that BD plants contained hydroxyl, carbonyl, and amine functional groups, double-bond aliphatic chains, various alkaloids, favonoids, polyphenols, terpenoids, and steroids [6].…”