基于UPLC-Q-Orbitrap HRMS技术的龙眼叶降血糖有效部位化学成分分析

Analysis and Identification of Chemical Constituents in Hypoglycemic Effective Fractions of Longan Folium Based on UPLC-Q-Orbitrap HRMS

目的:为明确龙眼叶的降血糖物质基础,采用超高效液相色谱-四级杆-静电场轨道阱高分辨质谱法(UPLC-QOrbitrap HRMS)对龙眼叶有效部位的化学成分进行快速分析和鉴定。方法:采用Thermo Hypersil GOLD C18色谱柱(2.1 mm×100 mm,1.9μm),流动相0.1%甲酸乙腈溶液-0.1%甲酸水溶液(含10 mmol乙酸铵)梯度洗脱,通过正、负离子模式进行扫描,扫描范围m/z 100~1500。结果:利用目标化合物的二级碎片离子信息与数据库及相关文献报道的化合物质谱信息进行比对,从乙醇部位中共鉴定出了9个化合物(木犀草苷、山柰酚、槲皮苷、木犀草素、莽草酸、柠檬酸、腺苷、烟酰胺和L-酪氨酸),乙酸乙酯部位中共鉴定出了11个化合物(木犀草苷、槲皮苷、山柰酚、木犀草素、莽草酸、没食子酸、原儿茶酸、腺苷、烟酰胺、东莨菪内酯和L-苯丙氨酸),正丁醇部位共鉴定出了10个化合物(木犀草苷、山柰酚-3-O-芸香糖苷、山柰酚、紫云英苷、木犀草素、柠檬酸、没食子酸、腺苷、烟酰胺和5-羟甲基糠醛),其中这3个活性部位的共有成分有5个(木犀草苷、山柰酚、木犀草素、腺苷和烟酰胺)。结论:建立的UPLC-Q-Orbitrap HRMS可快速鉴定龙眼叶中3个降血糖有效部位的化学成分。龙眼叶降血糖部位的主要化学成分为黄酮及其苷类、有机酸类和含氮类化合物。

Objective:Chemical constituents in hypoglycemic effective fractions of Longan Folium were isolated and identified by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Orbitrap HRMS)to clarify the hypoglycemic substance basis of Longan Folium.Method:Chemical constituents in hypoglycemic effective fractions of Longan Folium were isolated on a Thermo Hypersil GOLD C18 column(2.1 mm×100 mm,1.9μm),the mobile phase was 0.1%formic acid acetonitrile solution and 0.1%formic acid solution(containing and 10 mmol ammonium acetate)for gradient elution.HRMS was operated in the positive and negative ion modes with the scanning range of m/z 100-1500.Result:The secondary fragment ion information of target compounds was selected and compared with the compounds reported in the databases and related literature to further confirm these compounds.Nine compounds were identified in the ethanol fraction of Longan Folium,including cynaroside,kaempferol,quercitrin,luteolin,shikimic acid,citric acid,L-tyrosine,adenosine and nicotinamide.A total of 11 compounds were determined in the ethyl acetate fraction(cynaroside,quercitrin,kaempferol,luteolin,shikimic acid,gallic acid,protocatechuic acid,adenosine,nicotinamide,L-phenylalanine and scopoletin),and10 compounds were identified in the n-butanol fraction(cynaroside,kaempferol-3-O-rutinoside,kaempferol,astragalin,luteolin,citric acid,gallic acid,adenosine,nicotinamide and 5-hydroxymethylfurfural).And five common compounds were identified in these three hypoglycemic effective fractions.Conclusion:The established UPLC-Q-Orbitrap HRMS can quickly identify chemical constituents in three hypoglycemic effective fractions of Longan Folium,their main chemical constituents are flavonoids and their glycosides,organic acids and nitrogen-containing compounds,which provides technical support and scientific evidence for the study on pharmacodynamic material basis and quality control of Longan Folium.