基于超高效液相色谱-四极杆/静电场轨道阱高分辨质谱技术的复方血栓通胶囊化学成分研究

目的：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-Orbitrap HRMS）建立一种快速识别复方血栓通胶囊中多种化学成分的定性分析方法。方法：通过分析UHPLC-Q-Orbitrap HRMS技术捕捉复方血栓通胶囊中未知化合物的一级精确分子量和多级碎片信息，同时与对照品的保留时间和质谱数据信息进行比对，并结合相关参考文献以及Mass Bank、Chemical Book等网络数据库信息从而实现对未知物的准确快速定性。结果：共从复方血栓通胶囊中鉴定出52个化学成分，其中主要包括酚酸类、醌类、黄酮类、氨基酸类及其他类等化学成分。结论：建立的方法可系统、快速、准确地识别复方血栓通胶囊中的多种化学成分，并为其药效物质基础研究、质量控制及进一步临床应用等提供科学的理论依据。

Objective:To study the chemical composition of compound Xueshuantong capsules, and applied to a novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) for establishment of a method for recognization of the ingredients.Methods:The information of accurate mass in Full-MS and multistage fragment ions were obtained by the novel UHPLC-Q-Orbitrap technology.Chemical constituents were identified by comparing the relative retention time and the mass data of the reference substance, and consulting the reference literature or the Mass Bank, Chemical Book network database.Results:Fifty two compounds were identified in compound Xueshuantong capsules, including the phenolic acid, authraquinones, flavones, amino acids and others.Conclusion:A method is established to identify various chemical constituents in compound Xueshuantong capsules rapidly, accurately and systematically.And this research will lay a theoretical foundation and scientific study idea for the quality control improvement, bioactive components recognition and further clinical application of this herbal formulation.

[1] 孙志,胡玉荣,左莉华,等.UPLC-MS/MS法同时测定复方血栓通胶囊中9种成分的含量[J].中国药房,2017,28(21):2959 SUN Z,HU YR,ZUO LH,et al.Simultaneous determination of 9 components in compound Xueshuantong capsules by UPLC-MS/MS[J].China Pharm,2017,28(21):2959
[2] 龙克立.复方血栓通胶囊在眼底疾病治疗中的应用价值[J].中外医疗,2016,35(8):105 LONG KL.Application value of compound Xueshuantong capsule in treatment of fundus oculi disease[J].China Foreign Med Treat,2016,35(8):105
[3] 吴华锋,蔡雪桃,孔肖樱,等.复方血栓通胶囊治疗稳定型心绞痛的临床观察[J].亚太传统医药,2012,8(3):79 WU HF,CAI XT,KONG XY,et al.Clinical observation on the treatment of stable angina pectoris by compound Xueshuantong capsule[J].Asia Pac Tradit Med,2012,8(3):79
[4] 陈旭,周立新,田晓娟,等.工艺变更对中药制剂质量的影响[J].首都食品与医药,2015,22(14):5 CHEN X,ZHOU LX,TIAN XJ,et al.Influence of the quality for Chinese medicine preparation changed by process[J].Cap Food Med,2015,22(14):5
[5] 刘俊,朱宝平,孙志,等.基于超高效液相色谱-四极杆/静电场轨道阱高分辨质谱的妇可靖胶囊中11种成分定量研究[J].中草药,2018,49(2):353 LIU J,ZHU BP,SUN Z,et al.Quantitative research of multiple active components in Fukejing capsule based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry[J].Chin Tradit Herb Drugs,2018,49(2):353
[6] 渠琛玲,张寒琦,张华蓉,等.电喷雾质谱法研究氨基酸的质谱碎裂及其与人参皂苷Rb₃的相互作用[J].高等学校化学学报,2008,29(9):1721 QU CL,ZHANG HQ,ZHANG HR,et al.Studies on fragmentation pathways of amino acidsand their interactions with ginsenoside Rb₃ by pectrospray ionization mass spectrometry[J].Chem J Chin Univ,2008,29(9):1721
[7] 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 ZHANG Y,DONG WT,HUO JH,et al.Analysis on chemical constituents of Pheretima aspergillum by UPLC-Q-TOF-MS[J].Chin Tradit Herb Drugs,2017,48(2):252
[8] 秦伟瀚,刘飞,刘翔,等.基于UPLC-Q-TOF-MS法分析江西虫草化学成分[J].中药材,2017,40(6):1339 QIN WH,LIU F,LIU X,et al.Study on chemical compositions in Cordyceps jiangxiensis by UPLC-Q-TOF-MS[J].J Chin Med Mater,2017,40(6):1339
[9] National Center for Biotechnology Information.D-tert-leucine[EB/OL].USA,Bethesda:2006[2018-03-17]. https://pubchem.ncbi.nlm.nih.gov/compound/6950340#section=Top
[10] 王静哲,徐风,刘震,等.玄参化学成分HPLC-ESI-IT-TOF-MS分析[J].中国中药杂志,2016,41(7):1257 WANG JZ,XU F,LIU Z,et al.Identification of chemical constituents in Scrophulariae Radix by HPLC-IT-TOF-MS[J].China J Chin Mater Med,2016,41(7):1257
[11] 陶益,蒋妍慧,唐克建,等.地黄炮制前后化学成分的UHPLC-Q-TOF/MS比较研究[J].中药新药与临床药理,2016,27(1):102 TAO Y,JIANG YH,TANG KJ,et al.Comparison of chemical constituents of crude and processed products of Rehmannia glutinosa by using UHPLC-Q-TOF/MS[J].Tradit Chin Drug Res Clin Pharmacol,2016,27(1):102
[12] 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 SHU Y,SU SL,QIAN DW,et al.Chemical composition of Rougan capsule by UPLC-ESI-QTOF/MS[J].Chin Tradit Pat Med,2011,33(1):73
[13] 夏玮,古丽加玛丽·阿比斯,潘晨,等.腊梅花中黄酮类化合物的UHPLC/QTOF-MS分析[J].中成药,2014,36(11):2345 XIA W,GULIJIAMALI·A,PAN C,et al.Identification of flavonoids from Chimonanthus praecox(L.)Link by UHPLC/QTOF-MS[J].Chin Tradit Pat Med,2014,36(11):2345
[14] 陈平,路通,吴伯英,等.应用液相色谱-质谱联用技术检测血浆样品中丹参酚酸类成分的浓度[J].中国临床药理学与治疗学,2007,12(7):748 CHEN P,LU T,WU BY,et al.LC-MS/MS-based measurement of danshen phenolic acids in plasma[J].Chin J Clin Pharmacol Ther,2007,12(7):748
[15] 万建波,黎畅明,李绍平,等.加压溶剂提取与液相色谱-电喷雾质谱联用技术分析三七中皂苷类成分[J].中药材,2005,28(10):26 WAN JB,LI CM,LI SP,et al.Analysis of saponins from Panax notoginseng using pressurized solvent extraction coupled with liquid chromatography-electrospray mass spectrum[J].J Chin Med Mater,2005,28(10):26
[16] 徐文,丘小惠,张靖,等.超高压液相/电喷雾-LTQ-Orbitrap质谱联用技术分析三七根中皂苷类成分[J].药学学报,2012,47(6):773 XU W,QIU XH,ZHANG J,et al.Analysis of saponins in Panax notoginseng by UP LC-LT Q-Orbitrap MS/MS[J].Acta Pharm Sin,2012,47(6):773
[17] 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,1991,20(3):276 HUANG YS,FU JM,SHENG GY,et al.Identification and confirmation of α-cedrene and cuparene in high-vax oils and related Source rocks[J].Geochimica,1991,20(3):276
[18] 刘炎.中药黄芪炮制及复方配伍的化学研究[D].长春:长春中医药大学,2007 LIU Y.Study on Chemical Constituents of Processed Radix Astragali and Combination of Radix Astragali and Radix Angelicae Sinensis[D].Changchun:Changchun University of Chinese Medicine,2007
[19] 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 DONG X,XU L,LOU ZY.Determination of chemical components from Salvia miltiorrhiza Bunge by electrospray ion-trap mass spectrometry[J].Chin Pharm J,2010,45(14):1048
[20] 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 MAO Y,ZHANG RP,HE JH,et al.Determination of phenolic acids compounds in Arnebia euchroma(Royle)Johnstn.by Q exactive hybrid quadrupole-orbitrap mass spectrometer[J].Chin J Pharm Anal,2016,36(7):1199
[21] 张靖,丘小惠,徐文,等.基于液质联用技术的复方三芪口服液化学成分特征图谱研究[J].中华中医药杂志,2013,28(5):1572 ZHANG J,QIU XH,XU W,et al.Characteristic spectrum analysis of chemical components in Sanqi oral liquid by UPLC-MS/MS[J].Chin J Tradit Chin Med Pharm,2013,28(5):1572
[22] 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 KONG HN.The Identification of the Notoginseng Folium Saponins and the Purification of Notoginsenoside Fa[D].Tianjing:Tianjin University,2006
[23] 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 ZHU HM.Multicomponent Analysis and Pharmacokinetic Studies by UPLC/Q-TOF-MS/MS in Qiliqiangxin Capsules[D].Shijiazhuang:Hebei Medical University,2014
[24] National Center for Biotechnology Information.Ginsenoside Re[EB/OL].USA,Bethesda:2005[2018-03-17]. https://pubchem.ncbi.nlm.nih.gov/compound/441921
[25] 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):261 LIU J,YANG HH,LI XC,et al.Analysis of diterpene quinones in Danshen(Salvia miltiorrhiza Bunge)by HPLC-ESI/TOF MS[J].J Chin Mass Spectrom Soc,2008,29(5):261
[26] The Human Metabolome Database.Hydroxytanshinone(HMDB0036635)[EB/OL].Canada,Edmonton:2012[2018-03-12].http://www.hmdb.ca/metabolites/HMDB36635
[27] 闫海全.三七中主要皂苷类化合物的液相色谱分析与制备鉴定[D].成都:成都理工大学,2009 YAN HQ.The Analysis by HPLC,Isolation and Identification of Main Saponin Compounds in Panax notoginseng[D].Chengdu:Chengdu University of Technology,2009
[28] National Center for Biotechnology Information.Przewalskin[EB/OL].USA,Bethesda:2007[2018-03-17].https://pubchem.ncbi.nlm.nih.gov/compound/14139389#section=Top
[29] National Center for Biotechnology Information.Danshenxinkun B[EB/OL].USA,Bethesda:2006[2018-03-17].https://pubchem.ncbi.nlm.nih.gov/compound/5320113#section=Top
[30] National Center for Biotechnology Information.Neocryptotanshinone[EB/OL].USA,Bethesda:2005[2018-03-17].https://pubchem.ncbi.nlm.nih.gov/compound/184103#section=Top
[31] 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 KONG DY,LIU XJ,TENG MK,et al.The structure of Dan-Shen epiroketallactone of Dan-Shen(Salvia miltiorrhiza Bunge)[J].Acta Pharm Sin,1985,20(10):747
[32] 王红蕾,齐崴,吴小东,等.丹参水溶性多酚酸的多级串联电喷雾离子阱质谱断裂机理解析[J].质谱学报,2008,29(3):129 WANG HL,QI W,WU XD,et al.Fragmentation mechanisms of phenolic acids from Danshen in an ion trap by electrospray ionization multi-stage tandem mass spectrometry[J].J Chin Mass Spectrom Soc,2008,29(3):129