基于UHPLC-Q-Exactive Orbitrap HRMS和HPLC-UV的石吊兰化学成分定性与定量研究  

Qualitative and quantitative study of constituents in Lysionoti Herba based on UHPLC-Q-Exactive Orbitrap HRMS and HPLC-UV

作　　者：刘春花[1] 谢家丽 付昌丽 陆苑 潘洁[1] 刘亭 李勇军[1] 王永林 黄勇 孙佳 LIU Chun-hua;XIE Jia-li;FU Chang-li;LU Yuan;PAN Jie;LIU Ting;LI Yong-jun;WANG Yong-lin;HUANG Yong;SUN Jia(State Key Laboratory of Functions and Applications of Medicinal Plants,Engineering Research Center for the Development and Application of Ethnic Medicine and TCM(Ministry of Education),Guizhou Medical University,Guiyang 550004,China;Guizhou Provincial Key Laboratory of Pharmaceutics,Guizhou Medical University,Guiyang 550004,China;School of Pharmacy,Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学,民族药与中药开发应用教育部工程研究中心,省部共建药用植物功效与利用国家重点实验室,贵州贵阳550004 [2]贵州医科大学药学院,贵州贵阳550004 [3]贵州医科大学贵州省药物制剂重点实验室,贵州贵阳550004

出　　处：《中国中药杂志》2023年第13期3516-3534,共19页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(U1812403);中央引导地方科技发展专项(黔科中引地[2018]4006);贵州省高层次创新型人才项目(黔科合平台人才-GCC[2022]031-1)。

摘　　要：应用超高效液相色谱-四极杆静电场轨道阱串联质谱(UHPLC-Q-Exactive Orbitrap HRMS)联用技术对石吊兰化学成分进行系统分析,并针对其主要化学成分进行HPLC-UV含量测定。采用Synergi^(TM) Hydro-RP 100?色谱柱(2 mm×100 mm,2.5μm),以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱,流速0.2 mL·min^(-1),柱温40℃,质谱分析采用电喷雾电离源(ESI),正、负离子模式下分别采集数据;结合对照品保留时间、质谱离子信息以及数据库和文献中相关质谱数据信息进行比对,识别石吊兰中的化学成分;进一步采用HPLC-UV同时测定不同批次石吊兰中5个成分(新绿原酸、绿原酸、连翘酯苷B、毛蕊花糖苷、石吊兰素)的含量,检测波长为330 nm。在石吊兰中共鉴定出了84个化合物,包括27个黄酮类、20个苯乙醇苷类、5个氨基酸类、18个有机酸类、1个生物碱类、6个核苷类和7个其他类化合物。新绿原酸、绿原酸、连翘酯苷B、毛蕊花糖苷、石吊兰素5个成分在一定浓度范围内与峰面积的线性关系良好(r>0.999),各成分的线性范围分别为3.22~102.90、12.84~410.82、31.63~1 012.01、25.00~800.11、4.08~130.51μg·mL^(-1),仪器精密度、方法重复性和溶液稳定性均良好,平均加样回收率为97.31%~100.2%,RSD为0.95%~2.4%。贵州不同产地、不同批次石吊兰药材中5个成分的含量存在一定的差异,其中连翘酯苷B的平均含量最高。该研究建立的定性分析方法能快速、高效地鉴定石吊兰的化学成分,所建立的定量分析方法简便、快捷、准确,能同时测定石吊兰中5个成分含量,为石吊兰药材质量评价提供了科学依据。Ultra-high performance liquid chromatography-quadrupole-Exactive Orbitrap high resolution mass spectrometry(UHPLC-Q-Exactive Orbitrap HRMS) was employed to systematically analyze the chemical constituents in Lysionoti Herba,and high perfor-mance liquid chromatography-ultraviolet(HPLC-UV) to determine the content of main compounds.A Synergi^(TM) Hydro-RP 100 ? colu-mn(2 mm×100 mm,2.5 μm) was used for gradient elution with acetonitrile-0.1% aqueous formic acid as the mobile phase at a flow rate of 0.2 mL·min^(-1) and a column temperature of 40 ℃.MS and MS/MS were conducted with electrospray ionization(ESI) in both positive and negative modes.The chemical components in Lysionoti Herba were identified by comparison with the retention time and mass spectra of reference compounds and the relevant mass spectral data reported in MS databases and relevant literature.Furthermore,the content of five constituents(neochlorogenic acid,chlorogenic acid,forsythoside B,acteoside,and nevadensin) in different Lysiono-ti Herba samples was simultaneously determined by HPLC-UV at the wavelength of 330 nm.A total of 84 compounds were identified in Lysionoti Herba,including 27 flavonoids,20 phenylethanoid glycosides,5 amino acids,18 organic acids,1 alkaloid,6 nucleosides,and 7 others.The content of neochlorogenic acid,chlorogenic acid,forsythoside B,acteoside,and nevadensin showed good linear relationship(r0.999) with the peak area within certain concentration ranges,which were 3.22-102.90,12.84-410.82,31.63-1 012.01,25.00-800.11,and 4.08-130.51 μg·mL^(-1),respectively.The instrument precision,method repeatability,and solution stability all met requirement,and the average recovery rate was 97.31%-100.2%,with RSD ranging from 0.95% to 2.4%.The content of the five components varied among different Lysionoti Herba samples collected from different regions of Guizhou,and the average content of forsythoside B was the highest.The established qualitative method can rapidly and efficiently identify the chemical components of Lysionoti Her

关 键 词：石吊兰 UHPLC-Q-Exactive Orbitrap HRMS HPLC-UV 成分鉴定 含量测定 

分 类 号：R284.1[医药卫生—中药学]