超高效液相色谱-轨道阱高分辨质谱法对人参寡糖提取物与蓖麻凝集素120相互作用的研究

引用本文: 吴怡, 邵思梦, 李孟媛, 万茜淋, 张哲, 杨洪梅. 超高效液相色谱-轨道阱高分辨质谱法对人参寡糖提取物与蓖麻凝集素120相互作用的研究[J]. 分析化学, 2023, 51(1): 112-119. doi: 10.19756/j.issn.0253-3820.221420 shu

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

超高效液相色谱-轨道阱高分辨质谱法对人参寡糖提取物与蓖麻凝集素120相互作用的研究

吴怡 ^1, ,
邵思梦 ^1, ,
李孟媛 ^1, ,
万茜淋 ^2, ,
张哲 ^2, ,
杨洪梅 ^{1,
,}

1.
长春中医药大学东北亚中医药研究院, 长春 130117;
2.
长春中医药大学人参科学研究院, 长春 130117

通讯作者: 杨洪梅,E-mail:yanghm0327@sina.cn

基金项目:
国家自然科学基金项目(No.82074548)、吉林省科技厅中青年科技创新创业卓越人才(团队)项目(No.20220623026TC)和吉林省卫生技术创新项目(No.2021JC075)资助。

摘要: 利用超高效液相色谱-轨道阱高分辨质谱(UHPLC-Orbitrap-MS)结合链霉亲和素包被的96孔板的方法筛选人参复杂体系中蓖麻凝集素120(RCA 120)的寡糖结合剂。将RCA 120修饰的96孔板与溶菌酶修饰的96孔板分别与寡糖标准品孵育,对未结合的寡糖利用UHPLC-Orbitrap-MS进行分析,利用峰面积的变化表征寡糖是否与RCA 120发生相互作用。对影响亲和力的因素包括孵育时间、孵育温度和缓冲液等进行了优化。在优化条件下,对生晒参、红参和西洋参的寡糖提取物进行RCA 120结合剂的筛选,从生晒参中筛选出3种二糖化合物和2种三糖化合物,从西洋参中筛选出2种二糖化合物和2种三糖化合物,发现生晒参和西洋参中具有相同单糖残基的寡糖与RCA 120的结合强度均具有显著性差异。但是,从红参中未筛选到RCA 120结合剂,推测这是由红参炮制工艺导致的差异。本方法简单、快速、准确,可高通量地识别和筛选复杂体系中多个糖类组分与蛋白质的相互作用,为糖和蛋白质相互作用的研究提供了新方法。

关键词:

English

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).

Abstract: 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.

Key words:

Ultra-high performance liquid chromatography-mass spectrometry
/ Ginseng
/ Oligosaccharides
/ Ricinus communis agglutinin 120
/ Carbohydrate-protein interaction

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沈阳化工大学材料科学与工程学院沈阳 110142

超高效液相色谱-轨道阱高分辨质谱法对人参寡糖提取物与蓖麻凝集素120相互作用的研究

通讯作者: 杨洪梅,E-mail:yanghm0327@sina.cn

English

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

Corresponding author: YANG Hong-Mei, yanghm0327@sina.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

超高效液相色谱-轨道阱高分辨质谱法对人参寡糖提取物与蓖麻凝集素120相互作用的研究

通讯作者: 杨洪梅,E-mail:yanghm0327@sina.cn

English

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

Corresponding author: YANG Hong-Mei, yanghm0327@sina.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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