ISOLATION AND QUANTITY DETERMINATION OF EURYCOMANONE FROM Eurycoma longifolia BY LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY (LC-MS/MS)

Abstract

Using chromatography methods, we isolated the principal compound eurycomanone from Eurycoma longifolia J. Its chemical structure was determined on the basis of spectroscopic data (¹H-NMR, ¹³C-NMR, and ESI-MS). This compound was purified (purity >99.5%) using the Agilent 218 purification system. The compound was used as a standard for analyzing eurycomanone in six samples. The liquid chromatography-mass-spectroscopy/mass spectroscopy system was used (LC-MS/MS). The conditions are as follows: chromatography column EC-C₁₈ (100 × 2.1 mm, 2.7 μm); the mobile phase consisted of acetonitrile (A) and 0.1% formic acid aqueous solution (B); the gradient elution of 15–60% A at 0–5 min; the flow rate at 0.3 mL·min⁻¹; the sample injection volume at 1 μL; the column temperature at 35 °C; solvent: Me:H₂O = 70:30. The MS conditions are as follows: ESI ion source; positive ions; ionization source tempeature: 300 °C; precursor-product ion pairs for multiple-reaction monitoring were m/z 409.1 → 391.0. The results showed that Bac Giang Eurycoma longifolia J. has the highest eurycomanone content (3.1336 ± 0.0005 mg/g); meanwhile, Dack Nong Eurycoma longifolia J. has the lowest (0.1716 ± 0.0001 mg/g).