Countercurrent Chromatographic Separation of Lipophilic Ascorbic Acid Derivatives and Extract from Kadsura coccinea Using Hydrophobic Organic-Aqueous Two-Phase Solvent Systems

Abstract

Countercurrent chromatographic (CCC) separation of lipophilic ascorbic acid derivatives and the crude extract from Kadsura coccinea was performed using the type-J multilayer coil planet centrifuge with a hydrophobic organic-aqueous two-phase solvent system composed of n-hexane/ethyl acetate/ethanol/aqueous 0.1% trifluoroacetic acid at the volume ratio of 5:5:6:2. The lipophilic ascorbic acid derivatives were separated in the order of L-ascorbyl 2,6-dibutyrate, L-ascorbyl 6-palmitate and L-ascorbyl 6-stearate by eluting the lower phase as the mobile phase, and L-ascorbyl 2,6-dipalmitate was separated by eluting the upper phase at the opposite direction. The above solvent system was then applied to the CCC separation of the extract prepared from K. coccinea. With lower phase mobile, the extract was mainly separated into two peaks corresponding to lignans and triterpenoids accordingly. The HPLC analysis of the fractions showed that the former peak contained Kadsulignan N, Schizandrin H and Neokadsuranin as lignans, and the latter peak, Micranoic acid A, Neokadsuranic acid B and beta-Sitosterol as triterpenoids. The overall results indicate that the hydrophobic organic-aqueous two-phase solvent system used in the present studies was useful for the CCC separation of lignans and triterpenoids present in the natural products.

Keywords:

• Countercurrent chromatography
• Lignan
• Lipophilic ascorbic acid derivative
• Natural product
• Triterpenoid
• Two-phase solvent system

ACKNOWLEDGMENTS

This work was supported in part by grants from both of the Ministry of Education, Culture, Sports, Science and Technology of Japan and College of Pharmacy, Nihon University, Chiba, Japan.