State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau 999078, China.
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau 999078, China.
J Chromatogr A. 2018 Feb 9;1536:216-227. doi: 10.1016/j.chroma.2017.09.041. Epub 2017 Sep 22.
In the present study, a system was developed for the analysis of phenolic acids in Salvia miltiorrhiza using online comprehensive two-dimensional hydrophilic interaction chromatography and reversed-phase liquid chromatography coupled to a DAD detector and hybrid linear ion trap-Orbitrap mass spectrometry (HILIC×RP-DAD-ESI/HRMS/MS). The system was configured based on the combination of an XBridge Amide column (150mm×4.6mm, 3.5μm) and Accucore PFP column (50mm×4.6mm, 2.6μm) for the first and second dimensions, respectively. An additional LC pump was used to dilute the eluent from the first dimension to decrease its elution strength in the second dimension. A back-flush trap column was selected as an interface to make up for the loss of efficiency and resolution due to the online dilution. Under the optimized conditions, a total of 196 peaks of polar compounds were successfully separated and detected in Salvia miltiorrhiza using the developed online HILIC×RP system, which exhibited high orthogonality (73%). The online combination of HILIC and RP provides powerful separation capability for the analysis of polar compounds in samples with complex matrices.
在本研究中，建立了一种使用在线亲水相互作用色谱和反相液相色谱与 DAD 检测器和混合线性离子阱-Orbitrap 质谱（HILIC×RP-DAD-ESI/HRMS/MS）联用分析丹参中酚酸类物质的系统。该系统基于 XBridge Amide 柱（150mm×4.6mm，3.5μm）和 Accucore PFP 柱（50mm×4.6mm，2.6μm）的组合进行配置，分别用于第一维和第二维。使用额外的 LC 泵将第一维的洗脱液稀释，以降低第二维的洗脱强度。选择反冲洗捕集柱作为接口，以弥补由于在线稀释而导致的效率和分辨率的损失。在优化的条件下，使用开发的在线 HILIC×RP 系统成功分离和检测了丹参中 196 个极性化合物峰，该系统表现出高正交性（73%）。HILIC 和 RP 的在线组合为分析复杂基质样品中的极性化合物提供了强大的分离能力。
