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 邵思梦 李孟媛 万茜淋 张哲 杨洪梅

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

    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。

English


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  • 收稿日期:  2022-08-15
  • 修回日期:  2022-09-27
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