成分分析

基于MALDI-MSI技术的纹党根代谢物的空间分布特征研究*

  • 刘旭霞 ,
  • 刘晓玲 ,
  • 马海棠 ,
  • 王欣 ,
  • 陈正君 ,
  • 罗文蓉 ,
  • 杨扶德
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  • 1.甘肃中医药大学药学院,兰州 730000;
    2.甘肃省中医院药学部,兰州 730050
第一作者 Tel:18893796823;E-mail:gszylxx6823@163.com
**杨扶德 Tel:18152064502;E-mail:gszyyfd@163.com
罗文蓉 Tel:13893303264;E-mail:gszylwr@163.com

收稿日期: 2024-06-04

  网络出版日期: 2025-05-29

基金资助

*科技部国家重点研发计划项目(2018YFC1706305); 甘肃省科技计划项目(21JR1RA271); 2021年度甘肃高等学校产业支撑计划项目(2021CYZC-40); 甘肃省重点研发计划项目(23YFFA0069); 兰州市科技局校企合作项目(2023-RC-9)

Spatial distribution characteristics of metabolities in root of Wendang: based on MALDI-MSI*

  • LIU Xu-xia ,
  • LIU Xiao-ling ,
  • MA Hai-tang ,
  • WANG Xin ,
  • CHEN Zheng-jun ,
  • LUO Wen-rong ,
  • YANG Fu-de
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  • 1. College of Pharmacy, Gansu University of Chinese Medicine, Lanzhou 730000, China;
    2. Department of Pharmacy, Gansu Provincial Hospital of Traditional Chinese Medicine, Lanzhou 730050, China

Received date: 2024-06-04

  Online published: 2025-05-29

摘要

目的: 建立一种原位可视化分析纹党[素花党参Codonopsis pilosula Nannf. var. modesta(Nannf.)L. T. Shen]根各类次级代谢物空间分布特征的新方法,实现对次级代谢物的组织定位,为深入挖掘纹党提供参考。方法: 利用基质辅助激光解吸电离质谱成像(MALDI-MSI)技术对纹党根代谢物进行质谱成像分析。基质为DHB+/DHB-(30 mg · mL-1),基质流速20 μL · min-1,氮气流速5 L · min-1,喷嘴移速3 mm · s-1,喷嘴温度60 ℃,每个切片喷涂时长50 min,用正、负离子2种模式,正离子检测模式下的激光能量强度为60%,负离子检测模式下的激光能量强度为40%,检测质量范围为m/z 70~1 050,质量分辨率70 000,空间分辨率为50 μm,像素大小420 px×200 px。同时对代谢物进行通路富集分析。结果: 共检测到214个代谢物,同时可视化了40个代表性代谢物的空间分布特征,不同种类的次级代谢物在木栓层-韧皮部-木质部的分布模式差异较大,黄酮主要分布于木质部,生物碱、酚类和羧酸主要分布在木栓层和韧皮部,苯丙素和醌类主要分布于木栓层,氨基酸大量存在于韧皮部,核苷酸分布于整个根组织,指标性成分苍术内酯Ⅲ和紫丁香苷分布于木栓层,党参炔苷在木栓层和木质部中均有分布。通路富集分析显示代谢物显著富集于代谢途径、次生代谢物的生物合成、黄酮生物合成、氨基酸生物合成以及碳代谢等通路。结论: 本研究可视化了纹党根代谢物的空间分布特征,研究结果对纹党品质控制,药材的鉴定,有效成分的提取分离,代谢物的代谢规律提供了一定的理论支撑。

本文引用格式

刘旭霞 , 刘晓玲 , 马海棠 , 王欣 , 陈正君 , 罗文蓉 , 杨扶德 . 基于MALDI-MSI技术的纹党根代谢物的空间分布特征研究*[J]. 药物分析杂志, 2025 , 45(1) : 59 -71 . DOI: 10.16155/j.0254-1793.2024-0372

Abstract

Objective: To establish a new method for in situ visualization of spatial distribution characteristics of various secondary metabolites in the root of Wendang (Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen), so as to realize the tissular localization of secondary metabolites, and to provide a reference for the in-depth excavation of the Wendang. Methods: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used for the mass spectrometry imaging analysis of the root metabolites in Wendang. The matrix was DHB+/DHB- (30 mg · mL-1), the substrate flow rate was 20 μL · min-1, the nitrogen flow rate was 5 L · min-1, the nozzle movement speed was 3 mm · s-1, the nozzle temperature was 60 ℃, and the spraying time of each slice was 50 min. The laser energy intensity in positive ion detection mode was 60%, and that in negative ion detection mode was 40%. The detection mass range was m/z 70-1 050, the mass resolution was 70 000, the spatial resolution was 50 μm, and the pixel size was 420 px×200 px. At the same time, the pathway enrichment analysis was also carried out for the metabolites. Results: A total of 214 metabolites were detected, and the spatial distribution characteristics of 40 representative metabolites were visualized. The distribution patterns of different kinds of secondary metabolites in the cork - phloem - xylem varied considerably, with flavonoids mainly distributed in xylem, alkaloids, phenolics, and carboxylic acids mainly in the cork and phloem, phenylpropanoids and quinones mainly in the cork, amino acids were abundant in the phloem, and nucleotides were distributed throughout the root tissues, the indicator components atractylenolide Ⅲ and syringoside were distributed in the cork, and lobetyolin was distributed in both the cork and the xylem. Pathway enrichment analysis showed that metabolites were significantly enriched in metabolic pathways, biosynthesis of secondary metabolites, flavonoid biosynthesis, biosynthesis of amino acids, and carbon metabolism pathways. Conclusion: This study visualizes the spatial distribution characteristics of metabolites in the roots of Wendang. The results of this study can provide certain theoretical support for the quality control, the identification, the extraction and separation of active ingredients, and the metabolic pattern of Wendang.

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