超高效液相色谱-四极杆静电场轨道阱-高分辨质谱技术鉴定干鲜黄芪中的差异代谢物与生物碱类化合物

Identification of differential metabolites and alkaloids compounds in dried and fresh Astragalus membranaceus by ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high-resolution mass spectrometry

中文关键词:  黄芪  生物碱  代谢物组学

英文关键词:Astragalus membranaceus  alkaloids  metabolomics  ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high-resolution mass spectrometry

中文摘要:

      目的 采用超高效液相色谱-四极杆静电场轨道阱-高分辨质谱技术(ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high-resolution mass spectrometry, UPLC-QE-HRMS)对不同干鲜状态下黄芪代谢物进行系统分析, 并探究生物碱类物质的差异。方法 通过主成分分析和正交偏最小二乘法判别分析筛选差异代谢物, 并结合京都基因和基因组数据库通路进行富集分析。结果 干鲜黄芪中共鉴定出4771种代谢产物, 其中包括49种生物碱类物质, 分属吲哚类、吡啶类、异喹啉类、吡咯烷类、咪唑类、哌啶类、嘌呤类、二萜类、莨菪烷类、吲哚里西啶类、有机胺类、甾体类12类。以P<0.05、倍数变化>2或<0.5以及变量重要性投影大于1为标准, 筛选出含量具有显著差异的生物碱类化合物。其中α-卡茄碱在鲜黄芪中含量高, 而蝙蝠葛碱、2-羟基腺嘌呤、3-羟基-2-甲基吡啶、烟酰胺、L-缬氨酸以及促黑激素N-氧化物在干黄芪中含量高。结论 本研究揭示了干鲜黄芪中生物碱类化合物的差异, 为黄芪生物碱类活性成分的进一步开发利用提供了理论依据。

英文摘要:

      Objective To employ ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high-resolution mass spectrometry (UPLC-QE-HRMS) to systematically analyze the metabolites in Astragalus membranaceus under different dried and fresh states, explore the differences of alkaloid substances. Methods Differential metabolites were screened using principal component analysis and orthogonal partial least squares discriminant analysis, and enrichment analysis was conducted based on pathways from the Kyoto Encyclopedia of Genes and Genomes. Results Total of 4771 kinds of metabolites were identified in both dried and fresh Astragalus membranaceus, including 49 kinds of alkaloids belonging to 12 categories: Indoles, pyridines, isoquinolines, pyrrolidines, imidazoles, piperidines, purines, diterpenoids, tropanes, indolizidines, organic amines and steroids. Alkaloid compounds with significant different content were screened using the criteria of P<0.05, fold change>2 or <0.5, and variable importance in projection>1. Among these, α-calcaine was found in highercontent in fresh Astragalus membranaceus, whereas dauricine, 2-hydroxyadenine, 3-hydroxy-2-methylpyridine, nicotinamide, L-valine and N-oxide (melanocyte-stimulating hormone) were more abundant in the dried samples. Conclusion This study reveals the differences in alkaloid compounds between fresh and dried Astragalus membranaceus, providing a theoretical basis for the further development and utilization of its bioactive alkaloid components in this medicinal herb.