LC-PDA-MS/MS-Based Characterization of Key Phytochemicals in Eurycoma Longifolia Roots

Area of Science:

• Phytochemistry
• Natural Product Chemistry
• Pharmacognosy

Background:

• Eurycoma longifolia (E. longifolia) is a plant traditionally used for medicinal purposes.
• Understanding its chemical constituents is crucial for validating its therapeutic potential.
• Previous studies have identified various quassinoid glycosides in E. longifolia.

Purpose of the Study:

• To fractionate the crude extract of Eurycoma longifolia roots.
• To identify intense peaks and chemical compounds within these fractions.
• To investigate the presence of chromenones and chromones in E. longifolia for the first time.

Main Methods:

• Crude extract fractionation using column chromatography with six solvent systems of varying polarity.
• High-Performance Liquid Chromatography with Photodiode Array and Mass Spectrometry (HPLC-PDA-MS/MS).
• Ultra-Performance Liquid Chromatography coupled with Mass Spectrometry (UPLC-MS/MS) and Proton Nuclear Magnetic Resonance (¹H-NMR) for compound identification.

Main Results:

• Two fractions yielded nearly pure and intense peaks suitable for analysis.
• Putative identification of chromenone (coumarin) and chromone derivatives.
• Identification of known quassinoid glycosides (eurycomanone derived glycoside, 2,3-dehydro-4α-hydroxylongilactone glucoside, eurycomanol glycoside, eurycomanol trimer) in 100% methanol fraction.
• Proposed identification of a novel compound, hydroxyl glyyunanprosapogenin D (838 g/mol), in a 50% ethyl acetate/50% methanol fraction.

Conclusions:

• This study successfully fractionated E. longifolia root extract and identified key compounds.
• It is the first study to report the presence of chromenones and chromones in E. longifolia.
• The findings expand the known chemical profile of E. longifolia, including a potentially novel compound.