基于UPLC-Q-TOF-MS/MS研究法半夏中甘草化学成分
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摘要
目的:应用超高效液相色谱串联四级杆飞行时间质谱(UPLC-Q-TOF-MS/MS)对法半夏中所含甘草化学成分进行研究。方法:以法半夏水提液作为供试品溶液,采用Waters Acquity BEH C18色谱柱(2.1 mm×100 mm,1.7μm),乙腈-0.1%甲酸水为流动相进行二元梯度洗脱,流速0.25 m L·min-1,柱温35℃。质谱采用电喷雾离子源(ESI),负离子模式下扫描采集数据,根据准分子离子及二级碎片离子,并结合对照品及文献报道的质谱数据对法半夏中所含甘草化学成分进行鉴定。结果:从法半夏中鉴定出15个甘草化学成分。其中三萜皂苷类成分6个,黄酮类成分9个。结论:该研究通过UPLC-QTOF-MS/MS分析技术,明确了法半夏中含有的15种甘草化学成分,可以为阐明半夏经甘草炮制成为法半夏后功效发生变化的物质基础及其临床合理应用提供依据。
Objective: To analyze the chemical components of Glycyrrhizae Radix et Rhizoma in Pinelliae Rhizoma Praeparatum(PRP) by using ultra performance liquid chromatography-quadrupole-time of flightmass spectrometry(UPLC-Q-TOF-MS/MS). Method: The aqueous extracts of PRP were separated on Waters Acquity BEH C18column(2. 1 mm × 100 mm,1. 7 μm) with acetonitrile-0. 1% formic acid solution as mobile phase for gradient elution. The flow rate was 0. 25 m L·min-1and the column temperature was 35 ℃. Mass spectrometric data were obtained under electrospray ionization(ESI) negative mode and the constituents were identified according to the exact relative molecular weight and fragment ions reported in the references. Result:Fifteen constituents were identified from Glycyrrhizae Radix et Rhizoma in PRP,including 6 triterpenoid saponins and 9 flavonoids. Conclusion: Systematic research of the fifteen chemical constituents of Glycyrrhizae Radix et Rhizoma in PRP was performed firstly in this study,which could provide the basis for clarifying the pharmacodynamic material basis and rational clinical application of PRP.
Objective: To analyze the chemical components of Glycyrrhizae Radix et Rhizoma in Pinelliae Rhizoma Praeparatum(PRP) by using ultra performance liquid chromatography-quadrupole-time of flightmass spectrometry(UPLC-Q-TOF-MS/MS). Method: The aqueous extracts of PRP were separated on Waters Acquity BEH C18column(2. 1 mm × 100 mm,1. 7 μm) with acetonitrile-0. 1% formic acid solution as mobile phase for gradient elution. The flow rate was 0. 25 m L·min-1and the column temperature was 35 ℃. Mass spectrometric data were obtained under electrospray ionization(ESI) negative mode and the constituents were identified according to the exact relative molecular weight and fragment ions reported in the references. Result:Fifteen constituents were identified from Glycyrrhizae Radix et Rhizoma in PRP,including 6 triterpenoid saponins and 9 flavonoids. Conclusion: Systematic research of the fifteen chemical constituents of Glycyrrhizae Radix et Rhizoma in PRP was performed firstly in this study,which could provide the basis for clarifying the pharmacodynamic material basis and rational clinical application of PRP.
引文
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[22]韩洁,赵晶,田维毅,等.肠菌群体外转化对甘草药用成分及镇咳作用的影响[J].安徽医科大学学报,2012,47(4):422-424.
[23]李若洁,石倩,程彬峰,等.甘草酸协同麻黄碱的平喘作用机制研究[J].药物评价研究,2010,33(3):183-186.
[24]石倩,刘赜,李若洁,等.利用基因芯片技术阐述甘草酸协同麻黄碱的分子作用机制[J].药物评价研究,2010,33(4):85-90.
[25]张明发,沈雅琴.甘草及其提取物对呼吸系统的药理作用[J].现代药物与临床,2010,25(4):262-267.
[26]Kamei J,Nakamura R,Ichiki H,et al.Antitussive principles of Glycyrrhizae Radix,a main component of the Kampo preparation Bakumondo-to(Mai-men-dongtang)[J].Eur J Pharmacol,2003,469(1/3):159-163.
[2]苏彬,李书渊,陈艳芬,等.半夏及其炮制品镇咳祛痰作用的比较[J].广东药学院学报,2013,29(2):181-184.
[3]游秋云,王平.生半夏、法半夏水提物对小鼠镇静催眠作用的比较研究[J].湖北中医杂志,2013,35(3):3-5.
[4]曾颂,李书渊,吴志坚,等.半夏镇咳祛痰的成分-效应关系研究[J].中国现代中药,2013,15(6):452-455.
[5]吴皓,谈献和,蔡宝昌,等.半夏姜制对麻黄碱含量的影响[J].中国中药杂志,1996,21(3):157-158.
[6]Oshio H,Tsukui M,Matsuoka T.Isolation of lephedrine from“pinelliae tuber”[J].Chem Pharm Bull,1978,26(7):2096-2097.
[7]周斌.麻杏石甘汤药效物质基础研究及新药研制[D].天津:天津大学,2007.
[8]赵杰.麻黄类药对组成规律的基础研究-麻黄-甘草药对(I)[D].广州:南方医科大学,2012.
[9]ZHANG Q Y,YE M.Chemical analysis of the Chinese herbal medicine Gan-Cao(licorice)[J].J Chromatogr A,2009,1216:1954-1969.
[10]周燕,王明奎,廖循,等.甘草化学成分的高效液相色谱-串联质谱分析[J].分析化学杂志,2004,32(2):174-178.
[11]LI Y J,CHEN J,LI Y,et al.Screening and characterization of natural antioxidants in four Glycyrrhiza species by liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry[J].J Chromatogr A,2011,1218:8181-8191.
[12]XUE Q,WEI S,SHUAI J,et al.Separation and characterization of phenolic compounds and triterpenoid saponins in licorice(Glycyrrhiza uralensis)using mobile phase-dependent reversed-phase×reversed-phase comprehensive two-dimensional liquid chromatography coupled with masss pectrometry[J].J Chromatogr A,2015,1402:36-45.
[13]Mohamed A F,Andrea P,Ludger A,et al.Comparative metabolite profiling and fingerprinting of medicinal licorice roots using a multiplex approach of GC-MS,LC-MS and 1D NMR techniques[J].Phytochemistry,2012,76(4):60-72.
[14]Montoro P,Maldini M,Russo M,et al.Metabolic profiling of roots of liquorice(Glycyrrhiza glabra)from different geographical areas by ESI/MS/MS and determination of major metabolites by LC-ESI/MS and LC-ESI/MS/MS[J].J Pharm Biomed Anal,2011,54(3):535-544.
[15]WANG Y Y,HE S,CHENG X C,et al.UPLC-Q-TOFMS/MS fingerprinting of traditional Chinese formula Sijunzi Tang[J].J Pharm Biomed Anal,2013,80(3):24-33.
[16]ZHENG Y F,QI L W,ZHOU J L,et al.Structural characterization and identification of oleananetype triterpene saponins in Glycyrrhiza uralensis Fischer by rapid-resolution liquid chromatography coupled with time-of-flight mass spectrometry[J].Rapid Commun Mass Spectrom,2010,24(22):3261-3270.
[17]冷晶,朱云祥,陈璐琳,等.甘草中2个新三萜皂苷[J].中草药,2015,46(11):1576-1582.
[18]陶伟伟,段金廒,杨念云,等.乌拉尔甘草皂苷类成分研究[J].中草药,2013,44(12):1552-1557.
[19]王青,苗文娟,向诚,等.乌拉尔甘草中黄酮类化学成分的研究[J].中草药,2014,45(1):31-36.
[20]景晶,赵金英,华冰,等.甘草总黄酮抑制硫代乙酰胺诱导肝纤维化大鼠肝组织中TGF-β1及Caspase-3的表达[J].中国中药杂志,2015,40(15):3034-3040.
[21]俞腾飞,田向东,李仁,等.甘草黄酮、甘草浸膏及甘草次酸的镇咳祛痰作用[J].中成药,1993,15(3):32-33.
[22]韩洁,赵晶,田维毅,等.肠菌群体外转化对甘草药用成分及镇咳作用的影响[J].安徽医科大学学报,2012,47(4):422-424.
[23]李若洁,石倩,程彬峰,等.甘草酸协同麻黄碱的平喘作用机制研究[J].药物评价研究,2010,33(3):183-186.
[24]石倩,刘赜,李若洁,等.利用基因芯片技术阐述甘草酸协同麻黄碱的分子作用机制[J].药物评价研究,2010,33(4):85-90.
[25]张明发,沈雅琴.甘草及其提取物对呼吸系统的药理作用[J].现代药物与临床,2010,25(4):262-267.
[26]Kamei J,Nakamura R,Ichiki H,et al.Antitussive principles of Glycyrrhizae Radix,a main component of the Kampo preparation Bakumondo-to(Mai-men-dongtang)[J].Eur J Pharmacol,2003,469(1/3):159-163.