补气通络胶囊调控孕甾烷X受体活性成分的筛选及液相色谱-串联质谱同时测定研究  

Screening of Active Components of Pregnane X Receptor Regulated by Buqi Tongluo Capsules and Simultaneous Determination by Liquid Chromatography-Tandem Mass Spectrum

作　　者：黄楚燕 杨棣华 陈彭龙[1,2] 梁宏宇 李庆国 任辉[1,4] HUANG Chu-Yan;YANG Di-Hua;CHEN Peng-Long;LIANG Hong-Yu;LI Qing-Guo;REN Hui(Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Dept.of Pharmacy,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Dept.of Cardiovasology,Guangdong Second Traditional Chinese Medicine Hospital,Guangzhou 510095 Guangdong,China;Dept.of Orthopedics,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学,广东广州510405 [2]广州中医药大学第一附属医院药学部,广东广州510405 [3]广东省第二中医院心血管科,广东广州510095 [4]广州中医药大学第一附属医院骨科,广东广州510405

出　　处：《广州中医药大学学报》2022年第3期638-646,共9页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：广东省中医药管理局科研项目(编号:20211123)。

摘　　要：【目的】寻找补气通络胶囊中具有调控孕甾烷X受体(PXR)活性的成分,建立含量同时测定方法以评价该制剂的功效物质基础。【方法】根据补气通络胶囊内容物超高效液相色谱-飞行时间质谱(UPLC-TOF-MS)图谱及数据库分析结果,确定候选成分。将这些化合物的单体溶液加入PXR mRNA质粒转染的骨骼肌细胞中,孵育后采用聚合酶链式反应(PCR)法测定PXR mRNA表达,筛选出其中对比空白组产生明显变化且具有量效关系的化合物作为活性成分。以上述活性成分为指标,建立超高效液相色谱-串联质谱(UPLC-MS)含量同时测定的方法,根据结果选择活性、含量均适宜发挥功效的成分作为药效物质并评价其作用程度。【结果】补气通络胶囊中,有丹参酮ⅡA、欧当归内酯A、藁本内酯、川芎嗪、黄芪甲苷、毛蕊异黄酮葡萄糖苷、毛蕊花糖苷、人参皂苷Rb1、人参皂苷Rb3、人参皂苷Rd、人参皂苷Re、姜黄素等成分在UPLC-TOF-MS测定中信号较强且具有明显的PXR调控活性。UPHLC-MS含量测定结果显示,上述各成分除人参皂苷Rb3和人参皂苷Rd外,在全方制剂中的含量均符合发挥功效主治所需限度,可视为补气通络胶囊的药效物质。【结论】建立UPLC-TOF-MS含量同时测定的方法。补气通络胶囊中含有多种成分,主要涉及黄酮、皂苷、香豆素、萜类等。Objective To find the components with progesteroegnane X receptor(PXR)regulatory activity in Buqi Tongluo Capsules,and establish a simultaneous determination method to evaluate the efficacy of the preparationfunctional material basis.Methods The candidate constituents of Buqi Tongluo Capsules were determined by ultra-performance liquid chromatography-time-of-flight mass spectrum(UPLC-TOF-MS)and database analysis.The monomer solution of these compounds was added into the skeletal muscle cells transfected with PXR m RNA plasmids,and the PXR m RNA expression was determined by polymerase chain reaction(PCR)after incubation.The compounds with significant changes compared with the blank group and dose-effect relationship were selected as the active ingredients.Using the above active ingredients as indicators,a method for simultaneous determination of the content was established by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS).According to the results,the ingredients suitable for both activity and content were selected as pharmacodynamic substances and their effects were evaluated.Results In Buqi Tongluo Capsules,tanshinoneⅡA,levistilide A,ligustilide,ligustrazine,astragaloside,Calycosin-7-glucoside,verbascoside,ginsenoside Rb1,ginsenoside Rb3 ginsenosides RD,ginsenosides Re,curcumin and other components in the determination of UPLC-TOF-MS signal was stronger and had obvious activity of PXR regulation.UPHLC-MS content determination results showed that the contents of all the above ingredients except ginsenoside Rb3 and ginsenoside Rd in the whole preparation met the required limit of efficacy and indication,and could be regarded as medicinal substances of Buqi Tongluo Capsules.Conclusion A method for simultaneous determination of UPLC-TOF-MS is established.There are many components in Buqi Tongluo Capsules,involved in flavonoids,saponins,coumarins and terpenoids.

关 键 词：补气通络胶囊 孕甾烷X受体 超高效液相色谱-飞行时间质谱 质量控制 

分 类 号：R282.5[医药卫生—中药学]