High performance liquid chromatography in phytochemical analysis of Eurycoma longifolia

An analytical method using HPLC with UV detection was developed to investigate the quassinoid content of Eurycoma longifolio Jack (Simaroubaceae) collected from various sources. Eurycomanone (1), longilactone (2), 14,15 beta-dihydroxyklaineanone (3), 15 beta-acetyl-14-hydroxyklaineanone (4), 6 alpha-hydroxyeurycomalactone (5), and eurycomalactone (7) were isolated as reference standards and together with the synthesized 1 beta,12 alpha,15 beta-triacetyleurycomanone (6, internal standard), were identified by NMR, MS, UV and IR spectroscopies. Their coefficient of variation values for 0.50-35 mu g ml(-1) concentrations of quassinoids and their retention times measured within- and between-day were small. The recoveries of the spiked quassinoids in E. longifolio samples and their detection limits at 8.5 times signal to noise ratio were 99.75-109.13 % and 0.01 mu g ml(-1), respectively. From the root samples analysed, 1 had the highest concentration, being about 16.8-39.6 fold higher than the other quassinoids 2, 3, 5, 7 but 145.3 fold higher than 4 which showed the lowest concentration.