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On-line stop-flow two-dimensional liquid chromatography–mass spectrometry method for the separation and identification of triterpenoid saponins from ginseng extract

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Abstract

A method based on stop-flow two-dimensional liquid chromatography coupled with electrospray ionization mass spectrometry (2D LC-ESI MS) was established and applied to analyze triterpenoid saponins from the main root of ginseng. Due to the special structure of triterpenoid saponins (they contain polar sugar side chains and nonpolar aglycones), hydrophilic interaction chromatography (HILIC) and reversed-phase liquid chromatography (RPLC) were used for the two dimensions, respectively. A trap column was used to connect the two dimensions. The dilution effect, which is one of the main shortcomings of traditional comprehensive 2D LC methods, was largely avoided. The peak capacity of this system was 747 and the orthogonality was 56.6 %. Compared with one-dimensional HILIC or RP LC MS analysis, 257 and 185 % more mass spectral peaks (ions with intensities that were higher than 1,000) were obtained from the ginseng main root extracts, and 94 triterpenoid saponins were identified based on MSn information and summarized aglycone structures. Given its good linearity and repeatability, the established method was successfully applied to classify ginsengs of different ages (i.e., years of growth), and 19 triterpenoid saponins were found through statistical analysis to vary in concentration depending on the age of the ginseng.

Process of saponin's analysis using stop-flow 2D LC method

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Acknowledgments

The study was supported by the National Basic Research Program (nos. 2012CB720801, 2012CB517506) from the State Ministry of Science and Technology of China, the Foundations (nos. 21175132 and 21275141) and the Creative Research Group Project (no. 21321064) from the National Natural Science Foundation of China.

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Authors and Affiliations

  1. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China

    Shuangyuan Wang, Lizhen Qiao, Xianzhe Shi, Chunxiu Hu, Hongwei Kong & Guowang Xu

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  1. Shuangyuan Wang
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  2. Lizhen Qiao
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  3. Xianzhe Shi
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  4. Chunxiu Hu
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  5. Hongwei Kong
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  6. Guowang Xu
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Correspondence to Xianzhe Shi or Guowang Xu.

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Published in the topical collection Multidimensional Chromatography with guest editors Torsten C. Schmidt, Oliver J. Schmitz, and Thorsten Teutenberg.

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Wang, S., Qiao, L., Shi, X. et al. On-line stop-flow two-dimensional liquid chromatography–mass spectrometry method for the separation and identification of triterpenoid saponins from ginseng extract. Anal Bioanal Chem 407, 331–341 (2015). https://doi.org/10.1007/s00216-014-8219-4

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  • DOI: https://doi.org/10.1007/s00216-014-8219-4

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