基于超高效液相色谱-四级杆/静电场轨道阱高分辨质谱技术的活血通脉片化学成分研究
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摘要
目的:为系统研究复方中药制剂活血通脉片中的化学成分组成,建立采用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱(UHPLC-Q-Orbitrap HRMS)快速识别活血通脉片中多种化学成分的定性分析方法。方法:应用UHPLC-Q-Orbitrap MS技术捕捉未知化合物的精确分子质量及多级碎片离子信息,同时与标准品的相对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等网络数据库信息实现对未知物的准确快速定性。结果:共鉴定出59种化学成分,其中主要包括酚酸类、醌类、黄酮类、苯酞内酯类、皂苷类及其他类化学成分。结论:本研究建立的方法可系统、快速、准确地识别活血通脉片中的多种化学成分,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了科学的理论依据。
OBJECTIVE In order to investigate the chemical composition of Huoxuetongmai tablet, a novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was employed to establish a method that could identify the ingredients in Huoxuetongmai tablets systematically and rapidly. METHODS The information of accurate mass and multistage fragment ions were captured by advanced UHPLC-Q-Orbitrap technology. Chemical constituents were characterized by comparing the relative retention time and the mass data of the reference substance combined with the reference literature or the Mass Bank, Chemical Book and other network database, thus achieving accurate and rapid identification of unknown compounds. RESULTS Fifty-nine compounds were finally identified in this research, including the phenoliacids, anthraquinones, flavones, phthaleins and other categories. CONCLUSION A method was established in this study to identify various chemical constituents of Huoxuetongmai tablet systematically, rapidly and accurately. In addition, our work will lay a scientific foundation for the basic research of the bioactive components, quality control improvement and further clinical application of this herbal formulation.
OBJECTIVE In order to investigate the chemical composition of Huoxuetongmai tablet, a novel ultra high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was employed to establish a method that could identify the ingredients in Huoxuetongmai tablets systematically and rapidly. METHODS The information of accurate mass and multistage fragment ions were captured by advanced UHPLC-Q-Orbitrap technology. Chemical constituents were characterized by comparing the relative retention time and the mass data of the reference substance combined with the reference literature or the Mass Bank, Chemical Book and other network database, thus achieving accurate and rapid identification of unknown compounds. RESULTS Fifty-nine compounds were finally identified in this research, including the phenoliacids, anthraquinones, flavones, phthaleins and other categories. CONCLUSION A method was established in this study to identify various chemical constituents of Huoxuetongmai tablet systematically, rapidly and accurately. In addition, our work will lay a scientific foundation for the basic research of the bioactive components, quality control improvement and further clinical application of this herbal formulation.
引文
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[2] Hang QY, Xiang SL, Hu YC, et al. Intervention of Huoxue Tongmai Tablets on carotid vulnerable plaques [J]. Chin J Exp Clin Virol (中国实验方剂学杂志), 2017, 23(8):184-189.
[3] Sun XL, Lv YK. Clinical study on Huoxue Tongmai Tablets combined with trimetazidine in treatment of coronary heart Disease [J]. Drugs Clin (现代药物与临床), 2016, 31(9):1369-1373.
[4] Wang ZR, Xia T, Ma SP, et al. Effects of Huoxue Tongmai Tablet on ischemia-reper-fusion injury of myocardium in rats [J]. J Fourth Mil Med Univ (第四军医大学学报), 1999, 20(6):503-505.
[5] Hui TT, Xia ZT, Zhang LL, et al. Advances in study on effective eomponents of traditional Chinese medicine prescription [J]. Chin Med Herald (中国医药导报), 2012, 9(6):8-12.
[6] Zhao PJ, Zhang GP. Simultaneous determination of four active constituents in Huoxue Tongmai Tablets by HPLC [J]. Chin Tradit Pat Med (中成药), 2015, 37(12):2651-2655.
[7] Guo Q. A study on quality standard of Huoxue Tongmai Tablets [J]. Clin J Chin Med (中医临床研究), 2014, 6(16):8-11+16.
[8] Ju CL, Zhang HQ, Zhang HR, et al. Studies on fragmentation pathways of amino acids and their interactions with ginsenoside Rb3 by spectrospray ionization mass spectrometry [J]. Chem J Chin Univ (高等学校化学学报), 2008,29(9):1721-1726.
[9] Yu WW, Shuang PC, Zhang L. General research on chemical constituents and pharmacological activities of caulis spatholobi [J]. J Jiangxi Coll Tradit Chin Med (江西中医药大学学报), 2014, 26(4):89-92.
[10] Wu QR, Fang YQ, Wei G, et al. HPLC analysis on deoiled decoction of acorus gramineus [J]. Lishizhen Med Mater Med Res (时珍国医国药), 2002,13(07):385-387.
[11] Ding CG, Sun SQ, Zhou Q, et al. Chemical fingerprint analysis of tangerine peel reserved for different years by HPLC-DAD and HPLC-HRMS [J]. Chin J New Drug (中国新药杂志), 2008,17(11):927-930.
[12] PubChem, Salvianolic acid D[DB/OL], USA, 2014[2018-04-28], https://pubchem.ncbi.nlm.nih.gov/compound/75412558#section=Top.
[13] PubChem, Salvianolic acid G [DB/OL], USA, 2006 [2018-04-28], https://pubchem.ncbi.nlm.nih.gov/compound/11683160#section=Top.
[14] Liu J, Yang HH, Li XC, et al. Analysis of diterpene quinones in Danshen (Salvia Miltioeehiza Bunge) by HPLC-ESI/TOFMS [J]. J Chin Mass Spectr Soc (质谱学报), 2008, 29(5):261-267.
[15] Ying A, Yu QT, Guo L, et al. Structural-activity relationship of Ginsenosides from steamed Ginseng in the treatment of erectile dysfunction [J]. Ame J Chin Med, 2018, 46(1):137-155.
[16] Han W. Advances in chemical constituents and pharmacological effects of ligusticum chuanxiong [J]. Mod Chin Med (中国现代中药), 2017, 19(9):1341-1349.
[17] Cui YJ, Liu P, Chen RY, et al. Studies on the chemical constituents of spatholobus suberectus dunn [J] Acta Pharm Sin (药学学报), 2002, 37(10):784-787.
[18] Liao MC, Ge LL, Chen F, et al. Chemical constituents of acorus tatarinowii Schott [J]. J South-Central Univ Natl(Nat Sci Ed) (中南民族大学学报自然科学版), 2011, 30(04):45-47.
[19] PubChem, Salvianolic acid C [DB/OL], USA, 2007 [2018-08-26], https://pubchem.ncbi.nlm.nih.gov/compound/13991590#section=Top.
[20] Huang HL. Isolation and antioxidant activity of chemical constituents from salvia miltiorrhiza bunge [J]. Chin J Mod Drug Appl (中国现代药物应用), 2015, 9(21):277-278.
[21] Xv J, Jia L, Zhao YQ. Constituents of Ginseng and quality evaluation of Ginseng products [J]. Drug Eval Res (药物评价研究), 2011, 34(03):199-203.
[22] PubChem, Ginsenoside Rd1 [DB/OL], USA, 2015[2018-04-28], https://pubchem.ncbi.nlm.nih.gov/compound/102221467#section=Top.
[23] PubChem, Methylophiopogonanone A [DB/OL], USA, 2006 [2018-04-28], https://pubchem.ncbi.nlm.nih.gov/compound/5319741#section=Top.
[24] Zhang L, Ji J, Liu Y, et al. GC-MS analysis the constituents of volatile oils from Acorus tatarinowii Schott extracted by different methods [J]. J Liaoning Univ Tradi Chin Med (辽宁中医药大学学报), 2016, 18(01):53-55.
[25] PubChem, Tanshinone VI [DB/OL], USA, 2005 [2018-04-28], https://pubchem.ncbi.nlm.nih.gov/compound/149138.
[26] PubChem, Neocryptotanshinone [DB/OL], USA, 2005 [2018-04-28], https://pubchem.ncbi.nlm.nih.gov/compound/184103.
[27] Zhao Q, Li Y, Kong LS, et al. Research progress on chemical constituents and pharmacological activities of peach kernel [J]. J Tianshui Nor Univ (天水师范学院学报), 2008,(2):56-59.
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[29] Chi YM, Li Y, Zhang Y, et al. Analysis of flavonoids and components of stems & leaves of ranunculus ternatus thunb using ultra performance liquid chromatography-quadrupole tandem time of flight mass spectrometry [J]. Chin J Chromatogr (色谱), 2013,(9):838-844.
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[31] Gao LL. Studies on the chemical constituents and biological activity of Rheum Tanguticum Maxim. Ex Balf, Rheum Officinale Baill and Rheum Palmatum L.[D]. Peking Union Med Coll (北京协和医学院), 2012.
[32] Lin PF, Jia XZ, Qi Y, et al. Advances in study on phenolic Acids [J]. Guangdong Chem Ind (广东化工), 2017, 44(1):50-52.
[33] Wei L. Distribution and pharmacological effects of quinones [J]. Chin Med J Res Prac (现代中药研究与实践), 2013, 27(1):34-35,33.
[34] Shen ZY. Advances in antitumor activity of quinones and their derivatives [J]. Popul Sci Technol (大众科技), 2017, 19(10):55-56.
[35] Cao B, Wei GN. Research progress on pharmacological effects of flavonoids from Spatholobi Caulis [J]. Inter Med (内科), 2017, 12(3):341-343.
[36] Fang YJ, Xv YC, An MM, et al. Research advances in pharmacokinetics and pharmacological effects of flavonoids [J]. Pharm Care Res (药学服务与研究), 2015, 15(1):6-9.
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[38] Zheng SC, Li XY, Ou YB, et al. Research development on pharmacological of paeoniflorin [J]. Chin J Pharmacov (中国药物警戒), 2012, 9(2):100-103.
[39] Song Q. Research progress on chemical constituents of panax Ginseng and their pharmacological Effects [J]. Ginseng Res (人参研究), 2017, 29(2):47-54.