Quantitative Determination of Triperpene Saponins and Alkenated-Phenolics from Labisia pumila by LC-UV/ELSD Method and Confirmation by LC-ESI-TOF

Labisia pumila (Family, Myrsinaceae) is popularly known in Malaysia as kacip fatimah and is traditionally used as remedial for facilitation of labor, to delay fertility and regain body strength [1,2]. Until now, no thorough phytochemical investigation of this plant was available. Recently our group isolated triterpene saponins and alkenated phenolics from Labisia pumila which were utilized for method development.

This study describes the first analytical method for the determination of four triterpene saponins (Ardisicrenoside B (1), Ardisiacrispin A (2), 3-O-a-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-a-L-arabinopynanosyl) cyclamiretin A (3), Ardisimamilloside H (4)) and three alkenated-phenolics (Irisresorcinol (5) Belamcandol B (6), Demethylbelamcandaquinone B (7) in leaves of Labisia pumila. A separation by HPLC was achieved by using a reversed phase (RP-18) column, PDA with ELS detection, and a water/acetonitrile gradient as the mobile phase. The major triterpenoid (Ardisiacrispin A) and Irisresorcinol compounds were detected at a concentration as low as 10.0 and 1.0µg/mL respectively. The analysis of various samples showed considerable variation of 0.35–1.84% for the major terpenoid compound, ardisiacrispin A which is reported in Ardisia crispa for its utero contracting activities. LC-mass spectrometry coupled with electrospray ionization (ESI) method is described for the identification of seven compounds in plant samples. This method involved the use of the [M+Na]⁺ and [M+NH₄]⁺ ions for in the positive ion mode with extractive ion chromatogram (EIC).

Fig.1: HPLC-ELSD Chromatograms of a mixture of standard [Ardisicrenoside B (1), Ardisiacrispin A (2), 3-O-a-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-a-L-arabinopynanosyl) cyclamiretin A (3), Ardisimamilloside H (4), Irisresorcinol (5) Belamcandol B (6), Demethylbelamcandaquinone B (7)] (A), leaves of Labisia pumila (B, C), roots of Labisia pumila (D)

Acknowledgements: This research is supported in part by „Science Based Authentication of Dietary Supplements“ and „Botanical Dietary Supplement Research“ funded by the Food and Drug Administration grant numbers 5U01FD002071–09 and 1U01FD003871–01, and the United States Department of Agriculture, Agricultural Research Service, Specific Cooperative Agreement No. 58–6408–2-0009 and the authors would like to thank Annette Ford for the extractions of plant samples. References: [1] Ezumi MFW, et al. (2007) Indian J Pharmacol 39(1): 30–32. [2] Effendy AWM, et al. (2006)J Sustain Sci Manage 1(1): 40–46.