利用反相色谱-亲水作用色谱-预测多反应监测方法快速鉴定中药茵陈的化学成分组成
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摘要
阐明中药的化学成分组成是揭示其体内药效物质及作用机制的前提,也是中药化学生物学研究的重要组成。该研究以传统中药茵陈为例,探讨反相色谱-亲水作用色谱-预测多反应监测方法(RPLC-HILIC-p MRM)在中药化学成分快速发现、鉴定中的作用。通过将RPLC与HILIC直接耦联,整合两者的色谱分离特点,实现了大、中、小极性化学成分的全面保留和分离。中药中的绝大部分化学成分可以通过总结同属植物的化学成分研究结果,并引入初生代谢产物及可能的取代反应类型进行预测。因此,笔者针对可能的化学成分,构建了预测离子对,对茵陈中的化学成分进行快速识别和鉴定。通过与对照品比对、探讨裂解规律、对比文献和数据库信息等方式,共从茵陈中发现并初步鉴定了139个化合物。RPLC-HILIC-p MRM方法有效地避免了从海量的质谱数据中寻找有效信息,能够实现茵陈中化学成分的快速指认,可以作为中药化学成分分析的有效手段。
Chemical profiling of a given herbal medicine( HM) is the prerequisite for clarifying the effective material basis and therapeutic mechanisms,and it is an important integral part of traditional Chinese medicine chemical biology( TCMCB). In current study,we aimed to propose a new strategy for fast chemical characterization of HM by using reversed phase liquid chromatography-hydrophilic interaction chromatography-predictive multiple reaction monitoring( RPLC-HILIC-p MRM),and Artemisiae Scopariae Herba was employed in this study to illustrate the entire strategy. In response to wide polarity spanning of the diverse chemical clusters in Artemisiae Scopariae Herba,RPLC and HILIC were coupled in series to retain and separate hydrophilic and hydrophobic components simultaneously by identifying the characteristics of chromatographic separation. Most of the chemical constituents in traditional Chinese medicine can be predicted by summarizing the results of chemical constituents of the same genera and introducing primary metabolites and possible substitution reaction types. Therefore,we constructed predictive ion pairs to rapidly identify the chemical constituents of Artemisiae Scopariae Herba. After comparison with control products,discussion on fragmentation pattern,and access to relevant information from literature and databases,a total of 139 components were detected and structurally annotated by matching the obtained spectral data with the information of authentic compounds. Above all,RPLC-HILIC-p MRM could be used as an eligible analytical tool for the chemical profiling of HMs.
Chemical profiling of a given herbal medicine( HM) is the prerequisite for clarifying the effective material basis and therapeutic mechanisms,and it is an important integral part of traditional Chinese medicine chemical biology( TCMCB). In current study,we aimed to propose a new strategy for fast chemical characterization of HM by using reversed phase liquid chromatography-hydrophilic interaction chromatography-predictive multiple reaction monitoring( RPLC-HILIC-p MRM),and Artemisiae Scopariae Herba was employed in this study to illustrate the entire strategy. In response to wide polarity spanning of the diverse chemical clusters in Artemisiae Scopariae Herba,RPLC and HILIC were coupled in series to retain and separate hydrophilic and hydrophobic components simultaneously by identifying the characteristics of chromatographic separation. Most of the chemical constituents in traditional Chinese medicine can be predicted by summarizing the results of chemical constituents of the same genera and introducing primary metabolites and possible substitution reaction types. Therefore,we constructed predictive ion pairs to rapidly identify the chemical constituents of Artemisiae Scopariae Herba. After comparison with control products,discussion on fragmentation pattern,and access to relevant information from literature and databases,a total of 139 components were detected and structurally annotated by matching the obtained spectral data with the information of authentic compounds. Above all,RPLC-HILIC-p MRM could be used as an eligible analytical tool for the chemical profiling of HMs.
引文
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[15]付立忠,赵利梅,吕惠卿,等.氮素水平对三叶青生长和茎叶化学成分含量及抗氧化活性的影响[J].中国中药杂志,2019,44(4):696.
[2] Yan Y,Song Q Q,Chen X J,et al. Simultaneous determination of components with wide polarity and content ranges in Cistanche tubulosa using serially coupled reverse phase-hydrophilic interaction chromatography-tandem mass spectrometry[J]. J Chromatogr A,2017,1501:39.
[3] Huo H,Liu Y,Liu W,et al. A full solution for multi-component quantification-oriented quality assessment of herbal medicines,Chinese agarwood as a case[J]. J Chromatogr A,2018,1558:37.
[4] Song Q Q,Liu W J,Chen X J,et al. Serially coupled reversed phase-hydrophilic interaction liquid chromatography-tailored multiple reaction monitoring,a fit-for-purpose tool for large-scale targeted metabolomics of medicinal bile[J]. Anal Chim Acta,2018,1037:119.
[5] Jing W H,Yan R,Wang Y T. A practical strategy for chemical profiling of herbal medicines using ultra-high performance liquid chromatography coupled with hybrid triple quadrupole-linear ion trap mass spectrometry:a case study of Mori Cortex[J]. Anal Methods,2015,7(2):443.
[6] Yao C L,Pan H Q,Wang H,et al. Global profiling combined with predicted metabolites screening for discovery of natural compounds:characterization of ginsenosides in the leaves of Panax notoginseng as a case study[J]. J Chromatogr A,2018,1538:34.
[7]中国药典.一部[S]. 2015:239.
[8]欧阳文竹,尚展鹏,王文建,等.茵陈提取物中化学成分的UHPLC-LTQ-Orbitrap快速鉴定[J].中国中药杂志,2017,42(3):523.
[9]杨磊,刘梦娇,张志国,等.基于计算预测的茵陈蒿汤治疗肝硬化的“成分-靶标-通路”相关性分析[J].中国中药杂志,2018,43(7):1345.
[10] Wei R,Li G,Seymour A B. High-throughput and multiplexed LC/MS/MRM method for targeted metabolomics[J]. Anal Chem,2010,82(13):5527.
[11] Song Y L,Song Q Q,Li J,et al. An integrated strategy to quantitatively differentiate chemome between Cistanche deserticola and C. tubulosa using high performance liquid chromatography-hybrid triple quadrupole-linear ion trap mass spectrometry[J]. J Chromatogr A,2016,1429:238.
[12] Qian Z M,Li C H,Song Y L,et al. Rapid determination of adenosine in Cordyceps by online extraction HPLC[J]. J Chromatogr Sci,2019,doi:10. 1093/chromsci/bmz007.
[13] Luo P,Dai W,Yin P,et al. Multiple reaction monitoring-ion pair finder:a systematic approach to transform nontargeted mode to pseudotargeted mode for metabolomics study based on liquid chromatography-mass spectrometry[J]. Anal Chem,2015,87(10):5050.
[14] Zhang A,Sun H,Dou S,et al. Metabolomics study on the hepatoprotective effect of scoparone using ultra-performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry[J]. Analyst,2013,138(1):353.
[15]付立忠,赵利梅,吕惠卿,等.氮素水平对三叶青生长和茎叶化学成分含量及抗氧化活性的影响[J].中国中药杂志,2019,44(4):696.