SEPARATION AND PURIFICATION OF TWO MINOR COMPOUNDS FROMRADIX ISATIDISBY INTEGRATIVE MPLC AND HSCCC WITH PREPARATIVE HPLC.
Radix isatidis has been widely used as a Chinese traditional medicine for its anti-virus and anticancer activities where the minor components may contribute to these beneficial pharmaceutical effects. In order to enrich the target minor compounds effectively and rapidly, extraction, medium-pressure liquid chromatography (MPLC), high-speed countercurrent chromatography (HSCCC) and preparative high-performance liquid chromatography (pre-HPLC) were integratively used for separation and purification of two target minor compounds indole-3-acetonitrile-6-O-β-D-glucopyranoside (target 1) and clemastanin B (target 2) in the present study.Radix isatidiswas dried, pulverized and extracted with 50% methanol at room temperature, then concentrated and subjected to pretreatment with D-101 macroporous resin chromatography and extraction by MPLC. The first target compound was separated by MPLC at the purity raised to 70-80%, but without the second minor compounds which were irreversibly adsorbed by C18 solid support. Therefore, the second target compound in the crude extract was directly separated by HSCCC at purity of 80-90%. Finally these refined samples were further separated by pre-HPLC to obtain a high purity at 98-99%. The chemical structure identification of each target compound was carried out by IR, ESI-MS and1H NMR.
Institute of Analytical Chemistry, School of Chemistry and Environment, South China Normal University, Guangzhou, China.
Zhenjie
Z
Liang
Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518067, China.
Bin
B
Li
Institute of Analytical Chemistry, School of Chemistry and Environment, South China Normal University, Guangzhou, China.
Yong
Y
Liang
Institute of Analytical Chemistry, School of Chemistry and Environment, South China Normal University, Guangzhou, China.
Yaping
Y
Su
Laboratory of Bioseparation Technology, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA.
Yoichiro
Y
Ito
HPLC; HSCCC; MPLC; Macroporous resin; Radix isatidis