基于UHPLC-Q-Orbitrap HRMS技术的芪参益气滴丸中主要化学成分研究
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摘要
目的为更加全面、系统、快速地表征芪参益气滴丸有效化学成分谱,对芪参益气滴丸中主要化学成分进行快速识别鉴定和归属分类。方法采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q-OrbitrapHRMS)技术,ACQUITY UPLC?BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以乙腈(A)-0.1%甲酸水(B)为流动相进行梯度洗脱,以实现化合物的前期分离;通过先进的Q-Orbitrap MS技术捕捉未知成分的精确相对分子质量及多级碎片离子信息,同时与对照品的保留时间和质谱数据信息进行匹配,并结合相关文献以及ChemicalBook、MassBank等数据库最终实现对中药中复杂成分的快速准确识别。结果共鉴定出53种化学成分,其中包括有机酸类14种、黄酮类10种、醌类10种、皂苷类2种、氨基酸类7种及其他类10种。结论本法可快速、准确地识别芪参益气滴丸中主要化学成分,同时为其进一步的药效物质基础、质量控制及临床合理应用等研究奠定了科学的前期基础。
Objective To recognize the chemical profile of Qishen Yiqi Dropping Pills(QYDP) comprehensively, systematically and rapidly; ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was employed to identify the main chemical composition of this preparation. Methods A gradient elution for separation was achieved through ACQUITY UPLC? BEH C18 column(100 mm × 2.1 mm, 1.7 μm) with acetonitrile-water(containing 0.1% formic acid) as the mobile phase. Then, the novel Q-Orbitrap MS technology was employed to detect the information of accurate mass and multistage fragment ions. Chemical constituents were characterized by comparing their relative retention time and mass data with that of the reference substance, meanwhile the reference literature, Mass Bank database, and Chemical Book network database were also consulted. Results A total of 53 ingredients were finally identified in this study, including 14 organic acids, 10 flavones, 10 authraquinones, 2 saponins, 7 amino acids, and 10 others. Conclusion An efficient method was established in this work to identify the main chemical ingredients of QYDP rapidly and accurately; Meanwhile, this research will lay a scientific foundation for the further research on pharmacodynamic material basis, quality control, and clinical application of this medicine.
Objective To recognize the chemical profile of Qishen Yiqi Dropping Pills(QYDP) comprehensively, systematically and rapidly; ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was employed to identify the main chemical composition of this preparation. Methods A gradient elution for separation was achieved through ACQUITY UPLC? BEH C18 column(100 mm × 2.1 mm, 1.7 μm) with acetonitrile-water(containing 0.1% formic acid) as the mobile phase. Then, the novel Q-Orbitrap MS technology was employed to detect the information of accurate mass and multistage fragment ions. Chemical constituents were characterized by comparing their relative retention time and mass data with that of the reference substance, meanwhile the reference literature, Mass Bank database, and Chemical Book network database were also consulted. Results A total of 53 ingredients were finally identified in this study, including 14 organic acids, 10 flavones, 10 authraquinones, 2 saponins, 7 amino acids, and 10 others. Conclusion An efficient method was established in this work to identify the main chemical ingredients of QYDP rapidly and accurately; Meanwhile, this research will lay a scientific foundation for the further research on pharmacodynamic material basis, quality control, and clinical application of this medicine.
引文
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[35]毛艳,张瑞萍,贺金华,等.高效液相色谱-四极杆/静电场轨道阱高分辨质谱分析紫草中酚酸类化合物[J].药物分析杂志,2016,36(7):1199-1206.
[36]Wei W L,Huang L F.Simultaneous determination of ferulic acid and phthalides of Angelica sinensis based on UPLC-Q-TOF/MS[J].Molecules,2015,20(3):4681-4694.
[37]孔海宁.三七茎叶提取物中皂甙成分定性及三七皂甙Fa的制备[D].天津:天津大学,2006.
[38]汤喜兰,刘建勋,李磊.中药有机酸类成分的药理作用及在心血管疾病的应用[J].中国实验方剂学杂志,2012,18(5):243-246.
[39]姚梅芬,张思巨,李琳,等.苦参中黄酮类成分及其药理作用研究现状[J].中国中医药信息杂志,2013,20(3):110-112.
[40]魏蕾.醌类化合物的分布和药理作用[J].现代中药研究与实践,2013,27(1):34-35.
[41]傅晶,张东明,陈若芸.远志属植物的皂苷类成分及其药理作用研究进展[J].中草药,2006,37(1):144-146.
[2]陈景瑞,魏静,朱利洁,等.芪参益气滴丸对心血管系统疾病的药理作用研究进展[J].时珍国医国药,2015,26(2):434-437.
[3]王静,魏娜,赵洪芝,等.HPLC、UPLC测定芪参益气滴丸中5个丹参活性成分含量的比较研究[J].药物分析杂志,2011,31(9):1678-1682.
[4]孙志,周霖,姜晓芳,等.UPLC-MS/MS法结合模式识别同时测定芪参益气滴丸中11种成分[J].中成药,2017,39(10):2064-2068.
[5]龙辉,李怀国,刘东文.HPLC、UPLC测定芪参益气滴丸中丹参活性成分含量[J].深圳中西医结合杂志,2015,25(16):190-191.
[6]傅俊曾,宋生有,姜民,等.液相色谱-质谱联用同时测定芪参益气滴丸中黄芪甲苷、丹参素、原儿茶醛、人参皂苷Rg1和Rb1含量[J].中国药学杂志,2012,47(1):61-64.
[7]彭晶晶,李晓稳,李东翔,等.双波长超高效液相色谱指纹图谱鉴定芪参益气滴丸质量[J].中南药学,2016,14(11):1188-1193.
[8]陈慧贞.芪参益气滴丸多维指纹图谱研究[D].杭州:浙江大学,2012.
[9]李明,李瑞明,张依倩,等.芪参益气滴丸及其挥发性成分降香油的GC指纹图谱研究[J].中草药,2016,47(10):1702-1708.
[10]张玉,董文婷,霍金海,等.基于UPLC-Q-TOF-MS技术的广地龙化学成分分析[J].中草药,2017,48(2):252-262.
[11]束艳,宿树兰,钱大玮,等.柔肝胶囊化学成分的UPLC-ESI-QTOF/MS分析[J].中成药,2011,33(1):73-78.
[12]刘大伟,闫广利,方圆,等.UPLC-ESI-TOF/MS应用于黄芩化学成分的快速分析[J].中医药信息,2012,29(4):20-24.
[13]董昕,徐立,娄子洋.丹参药材中水溶性及脂溶性成分的电喷雾离子阱质谱研究[J].中国药学杂志,2010,45(14):1048-1054.
[14]赵洪芝,王静,姜民,等.丹参总酚酸提取物UPLC指纹图谱及成分定性研究[J].药物分析杂志,2012,32(4):620-622.
[15]Shu P,Hong J,Wu G.Analysis of flavonoids and phenolic acids in Iris tectorum by HPLC-DAD-ESI-MSn[J].Chin J Nat Med,2010,8(3):202-207.
[16]MassBank of North American,Baicalin[DB/OL].2018-03-12.http://mona.fiehnlab.ucdavis.edu/spectra/display/CCMSLIB00000078890.
[17]MassBank of North American,Ononin[DB/OL].2018-03-12.http://mona.fiehnlab.ucdavis.edu/spectra/display/PS086206.
[18]李东翔.液质联用法对黄芪浸膏入血成分的定性研究[A]//2012年中国药学大会暨第十二届中国药师周论文集[C].南京:中国药学会、江苏省人民政府,2012.
[19]孙进华,李先荣.UPLC-LTQ-Orbitrap快速鉴定丹参川芎嗪注射液中化学成分[J].中国实验方剂学杂志,2016,22(15):64-67.
[20]赵静,秦振娴,彭冰,等.基于UPLC-Q-TOF MS技术的三七中皂苷类成分质谱裂解规律研究[J].质谱学报,2017,38(1):97-108.
[21]朱慧明.基于液质联用技术的芪苈强心胶囊中多组分分析与药代动力学研究[D].石家庄:河北医科大学,2014.
[22]刘劼,杨黄浩,黎先春,等.高效液相色谱-电喷雾飞行时间质谱分析丹参中的丹参酮类化合物[J].质谱学报,2008,29(5):262-267.
[23]Human Metabolome Database,Hydroxytanshinone[DB/OL].2012-09-11/2018-03-12.http://www.hmdb.ca/metabolites/HMDB36635.
[24]PubChem,1beta-Hydroxycryptotanshinone[DB/OL].2006-10-25/2018-05-12.https://pubchem.ncbi.nlm.nih.gov/compound/10245003.
[25]Pub Chem,Przewalskin[DB/OL].2007-02-09/2018-05-12.https://pubchem.ncbi.nlm.nih.gov/compound/14139389.
[26]PubChem,Neocryptotanshinone[DB/OL].2005-08-09/2018-05-12.https://pubchem.ncbi.nlm.nih.gov/compound/184103.
[27]Pub Chem,Spathulenol[DB/OL].2005-08-08/2018-05-12.http://mona.fiehnlab.ucdavis.edu/spectra/display/PS087604.
[28]孔德云,刘星堦,滕脉坤,等.丹参中丹参螺旋缩酮内酯的结构[J].药学学报,1985,20(10):747-751.
[29]黄永松,傅家谟,盛国英,等.高蜡原油及其生油岩中新生物标志物-α雪松烯和花侧柏烯[J].地球化学,1991(3):276-282.
[30]MassBank of North American,Geranyl acetone[DB/OL].2018-03-12.http://mona.fiehnlab.ucdavis.edu/spectra/display/PS088901.2018.
[31]MassBank of North American,cis-Nerolidol[DB/OL].2018-03-12.http://mona.fiehnlab.ucdavis.edu/spectra/display/PS087604.2018.
[32]Masanori H,Toshisada S,Tsukasa I.New acylated anthocyanins and other flavonids from the flowers of clematis cultivars[J].Nat Prod Comun,2011,6(11):1631-1636.
[33]高亮亮.唐古特大黄、药用大黄和掌叶大黄的化学成分和生物活性研究[D].北京:北京协和医学院,2012.
[34]朱云祥,陈璐琳,龚婧如,等.LC-Q-TOF-MS及LC-IT-MS分析酸枣仁汤的化学成分[J].中国中药杂志,2014,39(3):457-465.
[35]毛艳,张瑞萍,贺金华,等.高效液相色谱-四极杆/静电场轨道阱高分辨质谱分析紫草中酚酸类化合物[J].药物分析杂志,2016,36(7):1199-1206.
[36]Wei W L,Huang L F.Simultaneous determination of ferulic acid and phthalides of Angelica sinensis based on UPLC-Q-TOF/MS[J].Molecules,2015,20(3):4681-4694.
[37]孔海宁.三七茎叶提取物中皂甙成分定性及三七皂甙Fa的制备[D].天津:天津大学,2006.
[38]汤喜兰,刘建勋,李磊.中药有机酸类成分的药理作用及在心血管疾病的应用[J].中国实验方剂学杂志,2012,18(5):243-246.
[39]姚梅芬,张思巨,李琳,等.苦参中黄酮类成分及其药理作用研究现状[J].中国中医药信息杂志,2013,20(3):110-112.
[40]魏蕾.醌类化合物的分布和药理作用[J].现代中药研究与实践,2013,27(1):34-35.
[41]傅晶,张东明,陈若芸.远志属植物的皂苷类成分及其药理作用研究进展[J].中草药,2006,37(1):144-146.