UHPLC-Q-Orbitrap HRMS鉴定丹灯通脑软胶囊中多种化学成分
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摘要
目的应用超高效液相色谱-四级杆/静电场轨道阱高分辨质谱(UHPLC-Q-Orbitrap HRMS)对丹灯通脑胶囊中的化学成分进行快速分析鉴别。方法采用ACQUITY UPLC?BEH C18色谱柱(2.1 mm×50 mm,1.7μm),乙腈-0.1%甲酸水为流动相进行梯度洗脱,分离各个化学成分,质谱采用全扫描模式,正负离子同时扫描检测色谱流出物。根据质谱提供的化合物精准分子量、多级碎片离子信息,结合相关参考文献和数据库信息,同时与对照品的相对保留时间和质谱数据进行比对,定性鉴定化合物。结果共鉴定出59种化学成分,主要包括黄酮类、醌类、有机酸类等化学成分,并对其药材来源进行了归属。结论利用UHPLC-Q-Orbitrap HRMS建立的鉴别方法,能够准确、系统、快速地鉴定丹灯通脑胶囊中的多种化学成分,为研究丹灯通脑胶囊药效物质基础及其质量标准提供依据和参考。
OBJECTIVE To identify the chemical constituents in Dandeng Tongnao capsule by ultra high performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry(UHPLC-Q-Orbitrap HRMS). METHODS The ACQUITY UPLC? BEH C18 chromatographic column(2.1 mm×50 mm, 1.7 μm) was used for UPLC, and the acetonitrile-0.1% formic acid water was used as mobile phase to separate various chemical constituents by gradient elution separation. Chromatographic effluent was detected by UHPLC-Q-Orbitrap HRMS full scan in positive and negative ion mode. According to precise molecular weight of compounds and multistage fragment ion information detected by UHPLC-Q-Orbitrap HRMS, reference and database information, it could be analyzed accurate characterization of the compound compared with relative retention time and mass spectrometry data of control sample. RESULTS Fifty-nine chemical constituents which mainly including flavonoids, quinones, and organic acids were identified and classified. CONCLUSION A variety of chemical constituents can be accurately identified by UHPLC-Q-Orbitrap HRMS. The results can be consulted by the further study of the pharmacodynamic basis and quality standard of Dandeng Tongnao capsule.
OBJECTIVE To identify the chemical constituents in Dandeng Tongnao capsule by ultra high performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry(UHPLC-Q-Orbitrap HRMS). METHODS The ACQUITY UPLC? BEH C18 chromatographic column(2.1 mm×50 mm, 1.7 μm) was used for UPLC, and the acetonitrile-0.1% formic acid water was used as mobile phase to separate various chemical constituents by gradient elution separation. Chromatographic effluent was detected by UHPLC-Q-Orbitrap HRMS full scan in positive and negative ion mode. According to precise molecular weight of compounds and multistage fragment ion information detected by UHPLC-Q-Orbitrap HRMS, reference and database information, it could be analyzed accurate characterization of the compound compared with relative retention time and mass spectrometry data of control sample. RESULTS Fifty-nine chemical constituents which mainly including flavonoids, quinones, and organic acids were identified and classified. CONCLUSION A variety of chemical constituents can be accurately identified by UHPLC-Q-Orbitrap HRMS. The results can be consulted by the further study of the pharmacodynamic basis and quality standard of Dandeng Tongnao capsule.
引文
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[2]国家食品药品监督管理局.国家中成药标准汇编:经络肢体、脑系分册[S].2002:254-257.
[3]李菲,郑国成,屈云萍,等.丹灯通脑胶囊剂的临床应用概况[J].中国民族民间医药,2016,25(13):35-37,41.
[4]QIU Y.Determination of puerarin and ferulic Acid in Dandengtongnao soft capsules by RP-HPLC[J].Anhui Med Pharm J(安徽医药),2013,17(2):212-213.
[5]SUN Z,JIANG X F,HU Y R,et al.Quantitative research on multiple active components in Dandeng Tongnao Soft Capsules by UPLC-MS/MS-pattern recognition[J].Chin Tradit Herb Drugs(中草药),2017,48(6):1126-1132.
[6]WANG P Y,WU C S,WU S,et al.Separation and characterization of 4 related impurities in puerarin bulk drug by HPLC-UV-FTICRMS/MSn[J].J Chin Mass Spectr Soc(质谱学报),2010,31(3):143-151.
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[9]Pub Chem.Caffeoylquinic acid[DB/OL].2004-09-16.http://pubchem.ncbi.nlm.nih.gov/compound/13991590.
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