UHPLC-Q-TOF/MS技术分析葛根醇提液中化学成分

Analysis of Chemical Constituents of Pueraria Lobata Alcohol Extract by UHPLC-Q-TOF/MS

目的采用超高效液相色谱-四级杆-飞行时间串联质谱(UHPLC-Q-TOF/MS)法对葛根醇提液中各成分进行分析和鉴定。方法色谱分离采用Poroshell 120 EC-C_(18)色谱柱(3.0 mm×100 mm,2.7μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相梯度洗脱,流速为0.4 mL·min^(-1);采用双电喷雾离子源(Dual ESI),在正负离子模式下采集数据;一级质谱提供精确的分子量信息,误差限制在5×10-6以内,得到该分子量最为精确的元素组成;二级质谱提供化合物的裂解碎片信息,将该化合物的特征碎片离子与相关数据库和相关文献中报道的相关化合物信息进行比对,推测其可能的化学结构。结果在葛根醇提液中发现71个成分,主要为异黄酮类成分,另包含有机酸和甾醇类等成分,其中8个化合物尚未有相关文献报道,结构需进一步确认。结论该研究丰富了葛根中成分化学结构信息,为葛根药效物质研究奠定了基础。

Objective To analyze and identify the chemical constituents of the alcohol extract of Radix Puerariae by uhra-high performance liquid chromatography-quadrapole-time of flight mass spectrometry （UHPLC-Q-TOF/MS）. Methods The chromatographic sepa ration was performed on an Agilent Poroshell 120 EC-C18 column （3.0 mm×100 mm, 2.7μm）, 0.1% formic acid （A） and acetonitrile （B） were used as the mobile phase for gradient separation, and the flow rate was set at 0.4 mL·min^-1. The constituents were detected in both positive and negative modes by Dual ESI-Q-TOF mass spectrometry. The MassHunter calculated the most accurate formula by the accurate molecular weight, and the error was limited within 5 ppm. The secondary mass spectrum provided information on the fragmentation of the compounds, and the characteristic fragment ions of the compounds were compared with the compound information reported in the relevant databases and related literature to deduce the possible chemical structures. Results A total of 71 compounds in the alcohol extract of Radix Puerariae were primarily identified, including isoflavonoids, organic acids and sterols; 8 out of them have not been reported and further validation is needed. Conclusion This study provides a foundation for the research of effective substance of Radix Puerariae.