基于UPLC-MS/MS建立同时测定自发性高血压大鼠主动脉血管内皮细胞中杜仲7个活性成分含量的方法  

Content determination of seven active components of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats by UPLC-MS/MS

作　　者：李梦婷 胡贺佳 金阳 陈艺 陈思颖 李月婷 黄勇 郑林 黄静 巩仔鹏 LI Meng-ting;HU He-jia;JIN Yang;CHEN Yi;CHEN Si-ying;LI Yue-ting;HUANG Yong;ZHENG Lin;HUANG Jing;GONG Zi-peng(Provincial Key Laboratory of Pharmaceutics in Guizhou Province,State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Provincial Engineering Research Center for Development and Application of Ethnic Medicine and Traditional Chinese Medicine,Ministry of Education,School of Pharmaceutical Sciences,Guizhou Medical University,Guiyang 550004,China)

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

出　　处：《中国中药杂志》2023年第13期3623-3632,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81703661,82160789);贵州医科大学优秀青年人才计划项目(2022-104);国家大学生创新创业计划项目(202010660023);2020年国家大学生创新创业计划项目(451)。

摘　　要：该文旨在采用UPLC-MS/MS技术建立同时测定自发性高血压大鼠(spontaneously hypertensive rats,SHR)主动脉血管内皮细胞中杜仲7个活性成分京尼平苷酸(genipinic acid,GA)、原儿茶酸(protocatechuic acid,PCA)、新绿原酸(neochlorogenic acid,NCA)、绿原酸(chlorogenic acid,CA)、隐绿原酸(cryptochlorogenic acid,CCA)、松脂醇二葡萄糖苷[(+)-pinoresinol di-O-β-D-glucopyranosid,PDG]、松脂醇单葡萄糖苷[(+)-pinoresinol 4′-O-β-D-glucopyranoside,PG]含量的方法。首先选取合格的SHR模型,采用结扎贴壁法分离、培养原代大鼠主动脉血管内皮细胞(VEC),并进行传代培养,鉴定成功后,以VEC为研究对象,采用超高效液相色谱-三重四极杆质谱联用仪(UPLC-MS/MS)建立同时测定VEC中GA、PCA、NCA、CA、CCA、PDG、PG等7个成分含量的方法,包括专属性、线性、基质效应、回收率、准确度、精密度和稳定性等。该文所建方法的最低定量限为0.97~4.95μg·L^(-1),准确度为87.26%~109.6%,提取回收率为89.23%~105.3%,基质效应为85.86%~106.2%,稳定性为86.00%~112.5%,能够满足测量的要求。表明该文建立了快速、准确地同时测定自发性高血压大鼠主动脉血管内皮细胞中杜仲7个活性成分含量的UPLC-MS/MS测定方法,为开展杜仲提取物7个活性成分的细胞药代动力学研究奠定基础。In the present study,the contents of seven active components [genipinic acid(GA),protocatechuic acid(PCA),neochlorogenic acid(NCA),chlorogenic acid(CA),cryptochlorogenic acid(CCA),(+)-pinoresinol di-O-β-D-glucopyranosid(PDG),and(+)-pinoresinol 4′-O-β-D-glucopyranoside(PG)] of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats(SHR) were simultaneously determined by ultra-high liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS).The qualified SHR models were selected.The primary aortic endothelial cells(VECs) of rats were separated and cultured by ligation and adherence,followed by subculture.After successful identification,an UPLC-MS/MS method for simultaneously determining the contents of GA,PCA,NCA,CA,CCA,PDG,PG in seven components of Eucommiae Cortex in VECs was established,including specificity,linearity,matrix effect,recovery,accuracy,precision and stability.The established method had the lo-west limit of quantification of 0.97-4.95 μg·L^(-1),accuracy of 87.26%-109.6%,extraction recovery of 89.23%-105.3%,matrix effect of 85.86%-106.2%,and stability of 86.00%-112.5%.Therefore,the established accurate UPLC-MS/MS method could rapidly and simultaneously determine the contents of the seven active components of Eucommiae Cortex in VECs of SHRs,which provided a refe-rence for the study of cellular pharmacokinetics of active components of Eucommiae Cortex extract.

关 键 词：杜仲提取物 血管内皮细胞 京尼平苷酸 松脂醇二葡萄糖苷 细胞药代动力学 

分 类 号：R285.5[医药卫生—中药学]