Purification of hydroxystilbenes from vine stalks cv. Chardonnay by centrifugal partition chromatography

Stilbenes naturally occur in several plant families, such as the Cyperaceae, Dipterocarpaceae, Gnetaceae, and Vitaceae [1,2]. Grapes (Vitaceae) and products manufactured from grapes are viewed as the most important dietary sources of these substances. Among stilbenes, hydroxystilbenes are of the most interest for their biological properties, such as potential cancer-chemopreventive [3] and neuroprotective activities [4].

Studies of their bioactivities (in vitro and in vivo tests), however, require large amounts of pure stilbenoids, which may be found in grapevine and particularly in stalk and stem extracts.

For this purpose, we investigated the purification of these polar compounds in a preparative-scale using centrifugal partition chromatography (CPC). This process differs from reversed HPLC in several ways. Indeed, higher selectivities can be attained by a skilled design of a solvent system, solvent usage can be significantly reduced, scale-up is easily achieved and there is no irreversible absorption on a solid support. In this study, we used successfully the quaternary Arizona solvent systems (n-heptane-EtOAc-MeOH-water) [5] for the purification of E-resveratrol, E-piceatannol, E-(ε)-viniferin, ampelopsin A, and E-vitisin C from stem of Vitis vinifera. Their anti-aggregating activity was evaluated against β-amyloid fibrils in vitro [6]. E-resveratrol and ampelopsin A were the most active compounds.

References: 1 Hart, J.H. (1981), Annu. Rev. Phytopathol., 19: 437–458. 2. Sotheeswaran, S., Pasupathy, V. (1993), Phytochemistry, 32:1083–1092. 3. Jang, M. et al. (1997), Science 275: 218–220. 4. Saiko, P., et al. (2008), Mut. Res. 658: 68–94. 5. Foucault, A.P., Chevolot, L. (1998)J. Chromatogr. A 808: 3–22