Quantitative Analysis of Eurycomanone in Eurycoma longjifolia Jack (Root) by High Performance Liquid Chromatography (HPLC)

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Jiranuch Mingmuang
Vilailak Chuennang-chee
Apirak Sakpetch
Somchit Niumsakul
Sakwichai Ontong
Nuchattra Chansuvanich


Eurycomanone is a quassinoid isolated from the roots of tongkat ali or pla-lai phueak (Eurycoma longjifolia Jack, Family Simaroubaceae). It has shown antipyretic, antimalarial, and aphrodisiac activities. Since there is no quality control of eurycomanone in tongkat ali root in Thailand, a HPLC method for determination of eurycomanone content has been developed and validated. Sample preparation was undertaken by refluxing 2 g of powdered drug in 50 ml of methanol for 1 hr, filtering and evaporating the extract until it was dry. The residue was redissolved and adjusted with 50% methanol to 10 ml. A 20-µl portion of sample solution was injected to HPLC system. The analysis was performed using Atlantis® T3, C18, 4.6 x 150 mm, 3-µm column; and solvent A, distilled water, and solvent B, a mixture of acetonitrile and methanol (1:1) were used as a mobile phase at a flow rate of 1.5 ml/min, and detected with PDA at wavelength 243 nm. Linearity was established for the eurycomanone concentration range of 0.06–0.36 mg/ml with a coefficient of determination (R2) 0.9995. The recovery rate was in the range of 96%–105%, while HORRAT was in the range of 1.66–1.96, and LOD and LOQ were 0.0106 µg/ml and 0.0354 µg/ml, respectively. It was shown that this developed method was suitable and could be used to analyze the content of eurycomanone in tongkat ali root and an appropriate specification of eurycomanone content in tongkat ali root could be established.


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