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Characterization of ginsenoside structural isomers from mixtures using in situ methylation with direct analysis in real-time ionization tandem mass spectrometry

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Abstract

Characterization of structural isomers of bioactive molecules is important for recognizing their functions, but it has been challenging due to their highly similar structures. As the main bioactive constituents of Panax ginseng, ginsenosides have different structural isomers attributed to the aglycone structure and glycosylation sites as well as stereochemistry of sugar groups attached. This work demonstrated a simple and robust in situ methylation reaction with tetramethylammonium hydroxide (TMAH) using ambient ionization source of direct analysis in real time (DART) to characterize saponin structural isomers. The DART ion source provides favorable conditions to methylate hydroxyl groups of ginsenoside instantaneously with TMAH, and it can ionize the methylated products at the same time. Methylated ginsenoside stereoisomers even with subtle structure differences generated very different mass signals from full-scan MS and tandem MS. High-resolution mass spectrometry aided the assignment of molecular structures of the various precursor and fragment ions from different ginsenosides, which provided structural information for both the aglycone skeleton and the sugar moieties in ginsenosides. The presented method was successfully used for the identification of ginsenosides in Panax ginseng, and saponin isomers were characterized without the need for chromatographic separation and/or tedious offline sample pretreatment.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China with Grant No. 21904067 and the Jiangsu Agricultural Science and Technology Innovation Fund, China (Grant No. SCX (20) 3083).

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Authors and Affiliations

  1. Jiangsu Collaborative Innovation Center of Biomedical Functional Materials, Jiangsu Key Laboratory of Biomedical Materials, School of Chemistry and Materials Science, Nanjing Normal University, 1 Wenyuan Road, Nanjing, 210023, China

    Rongfan Ren, Hongli Li, Qing Jiang & Xing Wang

  2. Department of Chemistry, University of British Columbia, Vancouver, BC, V6T 1Z1, Canada

    David Da Yong Chen

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Ren, R., Li, H., Jiang, Q. et al. Characterization of ginsenoside structural isomers from mixtures using in situ methylation with direct analysis in real-time ionization tandem mass spectrometry. Anal Bioanal Chem 415, 887–897 (2023). https://doi.org/10.1007/s00216-022-04482-w

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