基于UHPLC-Q-Orbitrap HRMS鉴定泽泻汤化学成分及小鼠体内成分组织分布特征  

Identifying chemical constituents of Zexie Decoction and its tissue distribution characteristics in mice by UHPLC-Q-Orbitrap HRMS

作　　者：陈森杰 高思琦 王星星 刘伟[2,3] 秦路平 吴建军 李荣胜 CHEN Senjie;GAO Siqi;WANG Xingxing;LIU Wei;QIN Luping;WU Jianjun;LI Rongsheng(College of Pharmaceutical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China;Grade Three Laboratory of Traditional Chinese Medicine Preparations of National Administration of Traditional Chinese Medicine,Department of pharmacy,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州310053 [2]上海中医药大学附属曙光医院药学部,国家中医药管理局中药制剂三级实验室,上海201203 [3]上海中医药大学附属曙光医院肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203

出　　处：《中草药》2024年第24期8336-8352,共17页Chinese Traditional and Herbal Drugs

基　　金：浙江省自然科学基金项目(ZCLY24H2801);浙江中医药大学“5151远志人才工程”(112123A12201/001/004/019);浙江中医药大学科研项目(2023JKZKTS20,2023RCZXZK20)。

摘　　要：目的借助超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q-Exactive Orbitrap HRMS)技术分析鉴定泽泻汤的化学成分,以及小鼠ig后主要入血、组织移行成分分布特征。方法取12只健康雄性C57/BL6J小鼠,分为3只空白组ig纯化水,9只给药组按21 g/kg ig泽泻汤提取液,分别在给药后0.5、1、3 h各处理3只,收集血清和组织样本,前处理采用蛋白沉淀法。采用ACQUITY UPLC BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以0.1%甲酸水溶液(A)-甲醇(B)为流动相体系,梯度洗脱分离。质谱采用正、负离子一级全扫描/数据依赖二级扫描模式(Full MS/dd-MS2)采集数据,综合离子峰相对保留时间、一级碎片离子信息,经Xcalibur 4.2软件拟合组成元素。通过对照品、文献以及数据库提供的二级碎片离子信息比对,实现对泽泻汤化学成分及入血、组织原型成分的鉴定。结果从泽泻汤中共鉴定出162种化学成分,泽泻127种、白术16种,共有成分19种;包括萜类128种(倍半萜类43种、三萜类82种、二萜类3种)、有机酸类12种、生物碱类7种、苯丙素类4种、黄酮类及苷类各2种、香豆素类1种以及其他类6种。小鼠ig泽泻汤后取样0.5、1、3 h检测到血清中60、56、45种;肺脏中46种、心脏中30种、脾脏中52种、肝脏中39种、肾脏中35种、回肠中61种、脂肪中46种原型成分。其中泽泻醇R、20-羟基泽泻醇C、泽泻醇O、泽泻醇C、白术内酯Ⅰ等14个化合物在血清和各组织中均有暴露。结论利用高分辨质谱技术完成了泽泻汤化学物质组及入血、组织移行成分的快速鉴定,可为泽泻汤效应物质的解析及药效成分在不同组织中的药代动力学研究提供参考。Objective UHPLC-Q-Exactive Orbitrap HRMS was used to analyze and identify the chemical constituents of Zexie Decoction(ZXD),as well as to characterize the distribution of the major blood-entry and tissue-migrating components in mice after gavage.Methods A total of 12 healthy male C57/BL6 J mice were divided into blank group(n=3)and treatment group(n=9).The blank group was given purified water by gavage,and the treatment group was given ZXD extract at 21 g/kg by gavage.Serum and tissue samples were collectedat 0.5,1 and 3 h after administration from three mice,respectively.The pretreatment was performed by protein precipitation method.The separation was performed on an ACQUITY UPLC BEH C_(18) column(100 mm×2.1 mm,1.7μm)with a mobile phase system of 0.1%formic acid aqueous solution(A)-methanol(B)by gradient elution.Mass spectrometry used positive and negative ion first-order full scan/data-dependent second-order scan mode(Full MS/dd-MS2)to collect data.The relative retention time of the ion peak and the first-order ion fragment information were integrated,and the constituent elements were fitted by Xcalibur 4.2 software.The identification of the chemical constituents and blood and tissue prototype components of ZXD was achieved by comparison of the control,literature and secondary fragment ion information provided by the database.Results A total of 162 chemical constituents were identified from ZXD,127 from Alisma plantago-aquatica,16 from Atractylodes macrocephala,and 19 common components.There were 128 kinds of terpenoids(43 kinds of sesquiterpenes,82 kinds of triterpenoids and three kinds of diterpenes),12 kinds of organic acids,seven kinds of alkaloids,four kinds of phenylpropanoids,two kinds of flavonoids and glycosides,one kind of coumarins and six kinds of others.Mice sampled 0.5,1,and 3 h after gavage of ZXD detected 60,56,and 45 components in serum,there were 46 prototypical components in lungs,30 in hearts,52 in spleens,39 in livers,35 in kidneys,61 in ileums,and 46 in fats.Among them,14 compounds such

关 键 词：UHPLC-Q-Exactive Orbitrap HRMS 经典名方 泽泻汤 入血成分 泽泻醇R 20-羟基泽泻醇C 泽泻醇O 泽泻醇C 白术内酯Ⅰ 组织分布 

分 类 号：R284.1[医药卫生—中药学]