Citation: WU Yi, SHAO Si-Meng, LI Meng-Yuan, WAN Xi-Lin, ZHANG Zhe, YANG Hong-Mei. Investigation of Interaction Between RCA120 and Ginseng Oligosaccharides by Ultra-High Performance Liquid Chromatography Coupled with Orbitrap Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2023, 51(1): 112-119. doi: 10.19756/j.issn.0253-3820.221420 shu

Investigation of Interaction Between RCA120 and Ginseng Oligosaccharides by Ultra-High Performance Liquid Chromatography Coupled with Orbitrap Mass Spectrometry

1.
Northeast Asia Institute of Chinese Medicine, Changchun University of Traditional Chinese Medicine, Changchun 130117, China;
2.
Jilin Ginseng Academy, Changchun University of Traditional Chinese Medicine, Changchun 130117, China

Corresponding author: YANG Hong-Mei, yanghm0327@sina.cn
Received Date: 15 August 2022
Revised Date: 27 September 2022

Fund Project: Supported by the National Natural Science Foundation of China (No.82074548), the Science and Technology Development Planning Project of Jilin Province, China (No.20220623026TC) and the Health Technology Innovation Project of Jilin Province, China (No.2021JC075).

A method was developed for screening protein binders from complicated ginseng samples by a streptavidin-coated 96-well plate format coupled with ultra-high performance liquid chromatography-orbitrap mass spectrometry (UHPLC-Orbitrap-MS). The Ricinus communis agglutinin 120 (RCA 120) coated 96-well plate and lysozyme coated 96-well plate (as control) were incubated with oligosaccharide standards respectively, and the compounds with the decreased peak areas in experimental group compared to those in the control group were detected as binders by UHPLC-ESI-MS. The factors such as incubation time, incubation temperature, and buffer, which might affect the binding affinity and reproducibility, were optimized. The potential of the approach was examined using the extracts of white ginseng, American ginseng and red ginseng. Three disaccharides and two trisaccharides were screened out from the extracts of white ginseng. As for the extracts of American ginseng, two disaccharides and two trisaccharides were observed to bind to RCA120. While no binders were detected in the extracts of red ginseng. Significant differences were observed in the relative binding degrees (RBDs) of the detected oligosaccharides in white ginseng and American ginseng binding to RCA 120. To our knowledge, it's the first time to reveal the differences and analogies in RCA 120-binding capabilities of oligosaccharides between the extracts of white ginseng and American ginseng, indicating the efficiency of the developed method for analysis of complicated samples.
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