中药药理与临床

目的:通过超高效液相色谱-电喷雾-四极杆-飞行时间-串联质谱(UPLC-QTOF-MS)技术研究降防保心胶囊入血成分，再建立降防保心胶囊的指纹图谱和含量测定方法。方法:通过UPLC-QTOF-MS技术考察降防保心胶囊入血成分。采用HPLC建立降防保心胶囊的指纹图谱，并利用《中药色谱指纹图谱相似度评价系统》(2012版),结合聚类分析(CA)、主成分分析(PCA)和偏最小二乘法判别分析(PLS-DA)对指纹图谱进行评价，并结合对照品图谱对色谱峰进行指认。根据指纹图谱建立的色谱条件，建立指标性成分含量同时测定的分析方法。结果:对大鼠血清分析得到87个化学成分，主要包括生物碱类、有机酸类、黄酮类、萜类、挥发油类等。通过HPLC建立降防保心胶囊的指纹图谱，结果显示，相似度>0.97。通过对照品指认了其中没食子酸、绿原酸、葛根素、芍药苷、粉防己碱、防己诺林碱、洋川芎内酯I、阿魏酸、苯甲酰芍药苷等9个共有峰。通过PCA、PLS-DA分析，8批样品分布比较集中，2批样品在默认核心区域外，并筛选出芍药苷、阿魏酸、绿原酸是降防保心胶囊成品的差异标志物。经方法学考察，建立芍药苷、阿魏酸、粉防己碱、防己诺林碱、葛根素、苯甲酰芍药苷、没食子酸的含量同时测定方法。结论:初步分析得出降防保心胶囊主要入血成分，并根据入血化学成分分析结果建立降防保心胶囊的指纹图谱和含量测定方法，所建方法准确、可靠，可用于降防保心胶囊的质量控制。

Objective:To analyze the plasma components of Jiangfang Baoxin(降防保心) Capsules(JFBX) by the ultra-high performance liquid chromatography-quadrupole time of flight tandem-mass spectrometry(UPLC-QTOF-MS),and to establish the fingerprint and content determination of JFBX. Methods:The plasma components of JFBX were investigated by UPLC-QTOF-MS. The fingerprint of JFBX was established by HPLC. Additionally, the fingerprint was evaluated by the Similarity Evaluation System of TCM Chromatographic Fingerprint(2012 edition) combined with cluster analysis(CA),principal component analysis(PCA),and partial least square discriminant analysis(PLS-DA). The chromatographic peaks were identified by combining the fingerprints of the reference substance. According to the chromatographic conditions established by fingerprint, an analytical method was established for the simultaneous determination of index components. Results:A total of 87 plasma components were obtained by UPLC-QTOF-MS,mainly including alkaloids, organic acids, flavonoids, terpenoids, volatile oils, and so on. The fingerprint of JFBX was established by HPLC,and the similarity was shown to be greater than 0.97. Additionally, 9 common peaks were identified by reference substance, including gallic acid, chlorogenic acid, puerarin, paeoniflorin, tetrandrine, fangchinoline, senkyunolide I,ferulic acid, and benzoyl paeoniflorin. Based on PCA and PLS-DA,8 batches of samples were closely distributed, the other 2 batches of samples were outside the default core area, with paeonidin, ferulic acid, and chlorogenic acid being screened as difference markers of JFBX. After methodological investigation, a method was established for the simultaneous determination of paeoniflorin, ferulic acid, tetrandrine, fangchinoline, puerarin, benzoyl paeoniflorin, and gallic acid. Conclusion:The plasma composition of JFBX is obtained by preliminary analysis. Then, the fingerprint and content determination method of JFBX are developed according to the analysis results. The established method is accurate and reliable and can be used for the JFBX quality control.