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Comprehensive two‐dimensional HPLC to study the interaction of multiple components in Rheum palmatum L. with HSA by coupling a silica‐bonded HSA column to a silica monolithic ODS column
Author(s) -
Hu Lianghai,
Li Xin,
Feng Shun,
Kong Liang,
Su Xingye,
Chen Xueguo,
Qin Feng,
Ye Mingliang,
Zou Hanfa
Publication year - 2006
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500442
Subject(s) - chromatography , monolithic hplc column , chemistry , high performance liquid chromatography , column (typography) , analytical chemistry (journal) , silica gel , two dimensional chromatography , chromatography detector , column chromatography , structural engineering , connection (principal bundle) , engineering
A mode of comprehensive 2‐D LC was developed by coupling a silica‐bonded HSA column to a silica monolithic ODS column. This system combined the affinity property of the HSA column and the high‐speed separation ability of the monolithic ODS column. The affinity chromatography with HSA‐immobilized stationary phase was applied to study the interaction of multiple components in traditional Chinese medicines (TCMs) with HSA according to their affinity to protein in the first dimension. Then the unresolved components retained on the HSA column were further separated on the silica monolithic ODS column in the second dimension. By hyphenating the 2‐D separation system to diode array detector and MS detectors, the UV and molecular weight information of the separated compounds can also be obtained. The developed separation system was applied to analysis of the extract of Rheum palmatum L., a number of low‐abundant components can be separated on a single peak from the HSA column after normalization of peak heights. Six compounds were preliminarily identified according to their UV and MS spectra. It showed that this system was very useful for biological fingerprinting analysis of the components in TCMs and natural products.