心脉安的UHPLC-Q-Exactive-Orbitrap-MS定性分析和UHPLC-QTRAP-MS/MS含量测定  

Identification of chemical constituents of Xinmai’an by UHPLC-Q-ExactiveOrbitrap-MS and determination of its content by UHPLC-QTRAP-MS/MS

作　　者：张薇 李楚源[1] 王德勤[1] 郭海彪[1] 孙晓波[2] 徐科一 殷玮 ZHANG Wei;LI Chu-yuan;WANG De-qin;GUO Hai-biao;SUN Xiao-bo;XU Ke-yi;YIN Wei(Hutchison Whampoa Guangzhou Bai Yunshan Chinese Medicine Co.,Ltd.,Guangzhou 510515,China;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Beijing 100193,China;Guangzhou General Pharmaceutical Research Institute Co.,Ltd.,Guangzhou 510240,China)

机构地区：[1]广州白云山和记黄埔中药有限公司,广东广州510515 [2]中国医学科学院药用植物研究所,北京100193 [3]广州医药研究总院有限公司,广东广州510240

出　　处：《中草药》2023年第6期1743-1756,共14页Chinese Traditional and Herbal Drugs

基　　金：名优中成药产学研深度融合发展服务平台(20212210006)。

摘　　要：目的应用UHPLC-Q-Exactive-Orbitrap-MS技术分析心脉安中的化学成分,同时建立一种快速同时测定心脉安中15种成分含量的UHPLC-QTRAP-MS/MS方法。方法采用ACQUITY UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm),0.1%甲酸水溶液(A)-乙腈(B)为流动相系统进行梯度洗脱,采用加热电喷雾离子源(HESI),正、负离子模式下采集数据,分析推定心脉安化学成分。并采用Waters ACQUITY UPLC CSH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),流动相为0.1%甲酸水(A)-乙腈(B),梯度洗脱,电喷雾离子源(ESI),采用多反应监测模式(MRM)正负离子同时对心脉安中15个成分进行定量分析。结果根据精确相对分子质量数据和多级质谱碎片离子,结合对照品比对或文献报道,共从心脉安中分析推定109个主要化学成分,包括31个三萜类(主要是三萜皂苷类)、25个黄酮类、15个苯丙素类、13个甾体、9个单萜类、7个醌类、9个其他类化合物。在优化的液相-质谱条件下,人参皂苷Re、人参皂苷Rg1、人参皂苷Rb1、人参皂苷Rd、丹参素、丹酚酸B、丹参酮I、隐丹参酮、丹参酮II_(A)、毛蕊异黄酮葡萄糖苷、黄芪甲苷、芍药苷、芍药内酯苷、麦冬皂苷D、鲁斯可皂苷元在测定浓度范围内线性关系良好(r≥0.9958),平均加样回收率(n=6)为91.8%~106.5%,RSD为1.9%~4.5%,精密度、稳定性、重复性试验的RSD均小于4.6%。结论该研究基本阐明了心脉安的化学物质基础,建立的UHPLC-QTRAP-MS/MS含量测定方法,重复性和专属性好,稳定可控,可用于心脉安的质量控制。Objective The UHPLC-Q-Exactive-Orbitrap-MS was developed for the identification of chemical constituents in Xinmai’an.and an UHPLC-QTRAP-MS/MS method was also established for rapid and simultaneous determination of the content of 15 components in Xinmai’an.Methods ACQUITY UHPLC HSS T3 chromatography column(100 mm×2.1 mm,1.8μm)was used with 0.1%formic acid(A)-acetonitrile(B)as the mobile phase in gradient elution.The data were collected by heated electrospray ion source(HESI)under positive and negative mode.Besides,Waters ACQUITY UPLC CSH C_(18)chromatography column(100 mm×2.1 mm,1.7μm)was used with 0.1%formic acid water(A)-acetonitrile(B)as mobile phase for gradient elution.Multiple reaction monitoring(MRM)in positive and negative ion mode.by using electrospray ion source(ESI)was performed for quantitative analysis of 15 compounds in Xinmai’an.Results Based on the accurate mass measurements,mass fragmentation patterns combined with comparison of reference or literature reports,a total of 109 major compounds including 31 triterpenoids(mainly triterpene saponins),25 flavonoids,15 phenylpropanoids,13 steroids,9 monoterpenes,7 quinones,and 9 other compounds were tentatively identified and speculated.Besides,under the optimized MS/MS condition,the linearity ranges of ginsenoside Re,ginsenoside Rg1,ginsenoside Rb1,ginsenoside Rb2,ginsenoside Rc,ginsenoside Rd,danshensu,salvianolic acid B,tanshinone I,cryptotanshinone,tanshinone II_(A),calycosin glucoside,astragaloside IV,paeoniflorin,albiflorin,ophiopogonin D,and ruscogenin were with good linearity(r≥0.9958).The average recovery rate of 15 compounds in the samples were in the range of 91.8%—106.5%,and the RSD range was from 1.9%to 4.5%.The RSD values of precision,stability and repeatability test were all less than 4.6%.Conclusion The results of this study clarified the chemical substance basis of Xinmai’an,and the method of UHPLC-QTRAP-MS/MS had good repeatability,high specificity,stability and controllability,which could be used for quality control of

关 键 词：心脉安 UHPLC-Q-Exactive-Orbitrap-MS UHPLC-QTRAP-MS/MS 三萜类 人参皂苷RE 人参皂苷Rg1 丹参酮Ⅰ 黄芪甲苷 芍药苷 

分 类 号：R284.1[医药卫生—中药学]