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Indexed by:期刊论文

Date of Publication:2014-07-18

Journal:JOURNAL OF CHROMATOGRAPHY A

Included Journals:SCIE、EI、PubMed、Scopus

Volume:1351

Page Number:90-95

ISSN No.:0021-9673

Key Words:Recycle preparative HPLC; Repetitive enrichment; Low abundance compound; Column capture; Mobile phase adjusting

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. (C) 2014 Elsevier B.V. All rights reserved.