基于UHPLC-Q-Exactive Orbitrap MS技术的血藤果化学成分分析

Identification of Chemical Constituents in the Kadsura Coccinea Fructus Based on UHPLC-Q-Exactive Orbitrap MS

目的建立分析血藤果化学成分的超高效液相色谱串联四极杆轨道阱质谱(UHPLC-Q-Exactive Orbitrap MS)法。方法色谱柱为Thermo Scientific Hypersil GOLD^(TM)aQ柱(100mm×2.1mm,1.9μm),流动相为乙腈(含0.1%甲酸)-0.1%甲酸溶液(梯度洗脱),流速为0.3mL/min,柱温为40℃,进样量为1μL;离子源为电喷雾(ESI)离子源,分别采用正、负离子扫描模式。结果采用一级母离子全扫描和数据依赖性前三强二级子离子扫描模式,通过数据库、对照品及参考文献共鉴定出51个化学成分,包括丝氨酸、L-精氨酸、天冬酰胺、谷氨酸等氨基酸类成分15个,柠檬酸、酒石酸、奎宁酸、莽草酸等有机酸类成分21个,生物碱、核苷等其他类成分15个。结论所建立的方法可高效、快捷地鉴定血藤果的化学成分,为后续药效物质和质量标准研究奠定了一定基础。

Objective To establish an ultra high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry(UHPLC-Q-Exactive Orbitrap MS)method for identify the chemical constituents in the fruit of Kadsura Coccinea Fructus.Methods The chromatographic column was Thermo Scientific Hypersil GOLD^(TM)aQ column(100 mm×2.1 mm,1.9μm),the mobile phase was acetonitrile(containing 0.1%formic acid)-0.1%formic acid solution(gradient elution),the flow rate was 0.3 m L/min,the column temperature was 40℃,and the injection volume was 1μL.The electrospray(ESI)ion source was adopted with two scanning modes of positive and negative ions respectively.Results Through the precursor ion full MS scan and the top three data-dependent MS2 scan mode,51 chemical components were identified by database,standards and references,including 15 amino acids such as serine,L-arginine,asparagine and glutamic acid,21 organic acids such as citric acid,tartaric acid,quinic acid and shikimic acid,and 15 other constituents such as alkaloids and nucleosides.Conclusion The established method can efficiently and quickly identify the chemical constituents of the Kadsura Coccinea Fructus,which lays a foundation for the study of subsequent pharmacodynamic substances and their quality standards.