FOOD SCIENCE ›› 2013, Vol. 34 ›› Issue (6): 27-32.doi: 10.7506/spkx1002-6630-201306006

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Large-Scale Isolation and Preparation of Soybean Saponin by High-Speed Countercurrent Chromatography Combined with Preparative HPLC

HUANG Yu-ai,YAN Ming-xia,ZHAO Da-yun   

  1. School of Agriculture and Biology,Shanghai Jiao Tong University, Shanghai 200240, China
  • Received:2011-10-07 Revised:2013-01-15 Online:2013-03-25 Published:2013-03-01

Abstract: The objective of this study is to isolate and purify soybean saponin from soybean germs by high-speed countercurrent chromatography (HSCCC) combined with preparative HPLC. The isoflavonoids were removed from crude extract by HSCCC first. The pure soybean saponin was then obtained by preparative HPLC with acetic acid or trifluoroacetic acid (TFA) as mobile phase in a gradient elution mode. The optimal conditions for preparative HPLC were dual mobile phases composed of acetonitrile containing 0.01% (V/V) TFA and millipore water at a flow rate of 10 mL/min for gradient elution. Five compounds in group A and two compounds in group B and 3 kinds of 2,3-dihydro-2,5-dinydroxy-6-methyl-4H-pyran-4-one (DDMP)- conjugated soybean saponins with high purity were prepared and their structures were elucidated by mass spectrum (MS) and nuclear magnetic resonance (NMR). Therefore, the developed method can be used to separate partially and fully acetylated soybean saponins as well as DDMP-conjugated and unconjugated soybean saponins in the original sample.

Key words: soybean saponin, high-speed countercurrent chromatography (HSCCC), preparative high performance liquid chromatography, mass spectroscopy (MS), nuclear magnetic resonance (NMR)

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