基于UPLC-Q-Exactive四极杆-静电场轨道阱高分辨质谱联用技术快速识别补肾活血方的化学成分

Rapid analysis on chemical constituents from Bushen Huoxue prescription by UPLC coupled with Q-Exactive quadrupole-Orbitrap MS

目的:利用UPLC-Q-Exactive四极杆-静电场轨道阱高分辨质谱仪联用技术,快速识别与鉴定补肾活血方中的化学成分。方法:采用ACQUITY UPLC BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱,质谱采用负离子监测模式、全扫描及自动触发二级质谱扫描功能,对补肾活血方化学成分进行快速识别及鉴定。结果:通过质谱数据、对照品比对共鉴定出58个化合物;结果显示,补肾活血方中的主要化学成分包括异黄酮类、酚酸类、人参皂苷类及少量的环烯醚萜类化合物。结论:该研究能够快速分析和确定补肾活血方的化学成分,为确定补肾活血方防治糖尿病性视网膜病变药效物质提供了依据。

Objective:To develop an ultra-high performance liquid chromatography couple with Q-Exactive quadrupole-Orbitrap mass spectrometry method for rapid recognition and identification of the chemical constituents from Bushen Huoxue prescription.Methods:The Bushen Huoxue prescription extract was separated on an ACQUITY UPLC BEH C18 column(100 mm×2.1 mm,1.7 μm),and eluted with a gradient of methanol-water containing 0.1% formic acid.The constituents from Bushen Huoxue prescription were rapidly recognized and identified by HRMS in the negative ion mode using both full scan and automatic triggering of the function of the two stage mass spectrometry scanning.Results:A total of 58 compounds were identified by referring to mass spectrometric data and comparing with reference substances.Bushen Huoxue prescription mainly contains isoflavones,phenolic acids,ginsenosides and a small quantity of iridoids.Conclusion:The research can be used to quickly analyze and determine chemical constituents from Bushen Huoxue prescription,which also provides an experimental basis for further study of effective components in the prevention and treatment of diabetic retinopathy.