基于UPLC-QE-Orbitrap-MS技术的芪蛭降糖胶囊化学成分分析

Analysis of Chemical Constituents in Qizhi Jiangtang Capsules Based on UPLC-QE-Orbitrap-MS

目的:为系统分析芪蛭降糖胶囊的化学组成,采用超高效液相色谱-四级杆-静电场轨道阱高分辨质谱法(UPLCQE-Orbitrap-MS)对该复方的化学成分进行鉴定。方法:采用ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8μm),以乙腈(A)-水(B)为流动相梯度洗脱(0~13 min,1%~25%A;13~21 min,25%~35%A;21~28 min,35%~85%A;28~30 min,85%~100%A;30~32 min,100%~1%A),流速设定0.2 mL·min-1,柱温30℃,进样量3μL;运用电喷雾离子源(ESI),扫描范围m/z100~1 500,正、负离子全扫描模式采集数据。采用对照品比对、自建化合物数据库搜索、文献信息比对、诊断离子筛选及Compound Discoverer 3.0软件推测的方法快速鉴定各离子峰。结果:共鉴定了芪蛭降糖胶囊中52个化学成分,并归属了化合物的来源药材,其中氨基酸类成分主要来源于水蛭,苯丙素类成分来源于黄芪和地黄,环烯醚萜苷类成分来源于地黄,香豆素和三萜类成分来源于黄芪,黄酮类成分来源于黄芪和黄精。结论:建立的UPLC-QE-Orbitrap-MS分析方法能全面快速地分析和鉴定芪蛭降糖胶囊中的化学成分,其中多种成分经现代药理研究证明具有改善糖尿病及其并发症相关症状的作用,体现了芪蛭降糖胶囊多成分协同作用的特点,可为其药效物质基础和质量控制研究提供参考。

Objective:To systematically analyze the chemical constituents of Qizhi Jiangtang capsules by ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry(UPLC-QE-Orbitrap-MS). Method: Analysis was conducted on a ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8 μm)with acetonitrile(A)-water(B)as the mobile phase for gradient elution(0-13 min,1%-25%A;13-21 min,25%-35%A;21-28 min,35%-85%A;28-30 min,85%-100%A;30-32 min,100%-1%A). The flow rate was 0.2 mL·min-1,the column temperature was 30 ℃,and the volume of sample injection was 3 μL. Electrospray ionization(ESI)was used to collect data in the negative and positive ion modes with the scanning range of m/z 100-1 500. Meanwhile,a variety of MS analytic methods were used,including comparing with the information of control substances,self-built compounds database and literature references,diagnostic ion filtering,Compound Discoverer 3.0 software,for identification of the chemical components.Result: Based on the above strategy,a total of 52 compounds were identified in Qizhi Jiangtang capsules,and the sources of these compounds were identified. Amino acids were mainly derived from Hirudo,phenylpropanoids were derived from Astragali Radix and Rehmanniae Radix,iridoid glycosides were derived from Rehmanniae Radix,coumarins and triterpenes were derived from Astragali Radix,flavonoids were from Astragali Radix and Polygonati Rhizoma. Conclusion: The established UPLC-QE-Orbitrap-MS analytical method can comprehensively and rapidly analyze and identify of the chemical constituents in Qizhi Jiangtang capsules. Many of the ingredients have been proved by modern pharmacological studies to have the effect of improving related symptoms of diabetes and its complications,reflecting the characteristics of synergistic action of multiple components in Qizhi Jiangtang capsules. This study can provide reference for the further research on the pharmacodynamic material basis and the quality control of Qizhi Jiangtang capsules.