基于UPLC-Q-Exactive Orbitrap-MS技术的车前子不同溶剂提取物成分分析

Analysis of the Chemical Constituents in Different Solvent Extracts of Plantaginis Semen by UPLC-Q-Exactive-Orbitrap Mass Spectrometry

目的:采用超高效液相色谱-四级杆-静电场轨道阱高分辨质谱联用技术(UHPLC-Q-Exactive Orbitrap-MS)分析车前子3种不同溶剂提取物的化学成分与差异,筛选车前子的最佳提取溶剂。方法:采用Waters ACQUITY UPLC BEH C色谱柱(100 mm×2.1 mm, 1.7μm),以0.1%甲酸水溶液(A)-甲醇(B)为流动相进行梯度洗脱,柱温30℃,流速为0.25 ml·min^(-1),进样量为2μl;质谱采用ESI源在正、负离子两种模式下采集数据。依据化合物精确质荷比及二级碎片离子信息,结合文献报导及各类化合物的一般裂解规律,分别对检测到的3种车前子提取液的化学成分进行鉴定和分析。结果:从车前子水提液、60%甲醇提取液、65%乙醇提取液中分别鉴定得到27,31,32个化学成分,包括黄酮类化合物11个、苯乙醇苷类化合物6个、环烯醚萜类化合物3个、三萜类化合物3个、生物碱类化合物3个等,其中差异性成分8个。结论:不同提取溶剂影响车前子提取物的成分组成,65%乙醇更适合作为车前子的提取溶剂。

Objective: To analyze and identify the chemical compositions and differences in three different solvent extracts of Plantaginis Semen by ultra-performance liquid chromatography coupled with hybrid quadrupole-Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS) in order to screen out the best extraction solvent.Methods: A Waters Acquity UPLC BEH Ccolumn(100 mm×2.1 mm, 1.7 μm) was used. The mobile phase of 0.1% formic acid in water and methanol was delivered at a flow rate of 0.25 ml·min^(-1) under a gradient program. The column temperature was 30℃ and the injection volume was 2 μl. The mass spectrometric data were acquired in both positive and negative ion modes using ESI. The components in three Plantaginis Semen extracts were identified by the high-precision mass-to-charge ratio and the secondary fragment ion information combined with literature reports and general law of cracking of various compounds. Results: Twenty-seven, thirty-one and thirty-two chemical constituents were identified from water extract, 60% methanol extract and 65% ethanol extract, respectively, including 11 flavonoids, 6 phenylethanol glycosides, 3 iridoids, 3 triterpenoids and 3 alkaloids, etc. Among them, there were 8 different components. Conclusion: According to the pharmacological actions, the amounts and contents of the obtained compounds, 65% ethanol is more suitable as the extraction solvent for Plantaginis Semen.