基于HPLC-Q-Exactive Orbitrap MS和特征分子网络技术快速鉴定滇白珠水提物化学成分

目的 采用高效液相色谱-四极杆-静电场轨道阱高分辨质谱（HPLC-Q-Exactive Orbitrap MS）结合特征分子网络（feature-based molecular networking，FBMN）技术，对滇白珠Gaultheria leucocarpa var. crenulata水提物化学成分进行快速分析。方法 采用Agilent Zorbax Eclipse XDB-C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈-0.1%甲酸水为流动相进行梯度洗脱，体积流量为0.8 mL/min，在正、负离子模式下，采集滇白珠水提物MS/MS数据，并在全球天然产物社会分子网络（Global Natural Products Social Molecular Networking，GNPS）网站（http://gnps.ucsd.edu）上创建FBMN。根据Compound Discoverer软件及GNPS数据库匹配结果，结合化合物的精确相对分子质量、特征碎片离子、色谱保留时间，参考对照品、文献及Pubchem、Massbank数据库，快速鉴定滇白珠水提物的化学成分。结果 共从滇白珠水提物中鉴定出130个成分，并区分了同分异构体，包括黄酮类40个、苯丙素类22个、糖苷类19个、有机酸类10个、生物碱类10个、酚类7个、萜类6个、酯类5个、氨基酸类4个、糖类3个、核苷酸类2个、醛类2个，其中5-O-咖啡酰奎宁酸、4-熊酰奎宁酸、邻苯二甲酸二丙酯这3个成分为根据FBMN结果推测解析。结论 FBMN弥补了传统分子网络在鉴定同分异构体方面的不足，HPLC-Q- Exactive Orbitrap MS结合FBMN能快速、准确、全面地鉴定滇白珠水提物化学成分，可为其进一步开发与应用提供依据。

Rapid identification of chemical components in water extract of Gaultheria leucocarpa var. crenulata based on HPLC-Q-Exactive Orbitrap MS and feature-based molecular networking technology

Objective To establish a method of high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry (HPLC-Q-Exactive Orbitrap MS) combined with feature-based molecular networking (FBMN) technology for rapidly analyzing chemical components in the water extract of Gaultheria leucocarpa var. crenulata. Methods Chromatographic separation was performed on an Agilent ZORBAX Eclipse XDB-C₁₈ column (250 mm×4.6 mm, 5 μm). Acetonitrile and 0.1% (volume fraction) formic acid solution were used as the mobile phase for gradient elution with the flow rate of 0.8 mL/min. FBMN was established on the Global Natural Products Social Molecular Networking (GNPS) website (http://gnps.ucsd.edu) based on MS/MS data of the water extract of G. leucocarpa var. crenulata, which was collected in positive and negative ion modes, respectively. According to the results of Compound Discoverer software and GNPS database, combined with accurate molecular weight, characteristic fragment ions, chromatographic retention time, standard substances, literatures, Pubchem and Massbank databases, chemical components in the water extract of G. leucocarpa var. crenulata were identified. Results A total of 130 components in the water extract of G. leucocarpa var. crenulata were identified and isomers were distinguished. These compounds included 40 flavonoids, 22 phenylpropanoids, 19 glycosides, 10 organic acids, 10 alkaloids, seven phenols, six terpenoids, five esters, four amino acids, three saccharides, two nucleotides and two aldehydes. Among them, three components, 5-O-caffeoylquinic acid, 4-p-coumaroylquinic acid, dipropyl phthalate, were inferred from the results of FBMN. Conclusion FBMN makes up for the deficiency of traditional molecular networking in the identification of isomers, HPLC-Q-Exactive Orbitrap MS combined with FBMN can quickly, accurately and comprehensively identify the components of the water extract of G. leucocarpa var. crenulata, and provide theoretical reference for its further development and utilization.