title: A novel preparative liquid chromatograph for repetitive enrichment and purification of low-abundance compounds.
abstract: A novel recycle preparative HPLC system was developed to repeatedly enrich and purify low-abundance compounds from complex samples. The recycle separation involves injecting a large volume of the sample through a trap column, releasing the absorbed sample for preparative HPLC separation, and then capturing the target component in the same trap column for the next separation. After several cycles, the final purified target compound absorbed in the trap column is eluted with a small volume of solvent. The capture and recycle procedure was realized by adjusting the strength of the mobile phase using an auxiliary pump and switching the two valves connecting the trap and preparative columns. Compared with standard HPLC or existing recycle chromatography, this system not only reduces the solvent side-effects and sample volume overloading during injection but also avoids peak-broadening and sensitivity loss during the recycle runs. Moreover, the final purified products can be easily concentrated. With the aid of this system, a 10% polyphenol in extracts from Saussurea involucrata cultured cells reached >95% purity with more than 95% recovery after three recycle purifications, and a 1% unstable epimer impurity in the raw material of the drug silybin reached >95% purity with 75% recovery after a six-cycle separation.
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China; Department of Bioscience and Biotechnology, School of Life science and Biotechnology, Dalian University of Technology, 2 Linggong Road, Dalian 116024,China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zou
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China.
authors.forename: Mingzhe
authors.initials: M
authors.lastname: Gao
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China.
authors.forename: Xiaozhe
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Bioscience and Biotechnology, School of Life science and Biotechnology, Dalian University of Technology, 2 Linggong Road, Dalian 116024,China.
authors.forename: Zhilong
authors.initials: Z
authors.lastname: Xiu
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, China; Institute of Chinese Materia Medica, Chinese Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: hbxiao@dicp.ac.cn.
authors.forename: Hongbin
authors.initials: H
authors.lastname: Xiao
mesh_terms: D002621:Chemistry / Q000295:instrumentation* / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D059808:Polyphenols / Q000302:isolation & purification; D030441:Saussurea / Q000737:chemistry; D012997:Solvents / Q000737:chemistry
chemical_list: D059808:Polyphenols; D012997:Solvents
keywords: Column capture; Low abundance compound; Mobile phase adjusting; Recycle preparative HPLC; Repetitive enrichment