Highly pure alpha-hederin from Hedera helix leaves isolated by centrifugal partition chromatography

Abstract

Introduction: Hedera helix is a species of the Araliaceae family, which contains flavonoids, coumarins, phenolic acids, sterols, alkaloids, anthocyanins, and triterpene saponins. a -Hederin is a saponin compound of the H. helix that possesses many potential biological activities. In previous studies, several methods were used to isolate and purify a -hederin from ivy leaves. However, the loss of a -hederin obtained during solid chromatography purification was relatively high, and it was difficult to obtain a -hederin with high purity. In this research, a centrifugal partition chromatogra-phy(CPC)methodwasestablishedfortheisolationof a -hederindirectlyfromethanolcrudeextract of Hedera helix leaves from Da Lat, Lam Dong province, Vietnam. Methods : The separation of a - hederin was carried out by applying centrifugal partition chromatography with a PLC 2250 system (Gilson Inc., France) coupled with an SCPC-250 column (Armen Instrument, France). In a CPC run, a two-phase solvent system consisting of n-hexane–ethyl acetate–methanol–water (2:3:2:3, v/v) in ascending mode (the lower phase was the stationary phase), with a rotation speed of 1600 rpm and a flow rate of 8.0 mL/min, was used. Results : The purity of the isolated a -hederin was 95.7%, as confirmed by HPLC-DAD at 210 nm. The structure of purified a -hederin was determined by 1 H-NMR and 13 C-NMR. The yield of a -hederin was 12.7%. Conclusion : This investigation might be used as a reference for the large-scale separation of high-purity a -hederin from H. helix leaves .