Pulsed elution modulation for on-line comprehensive two-dimensional liquid chromatography coupling reversed phase liquid chromatography and hydrophilic interaction chromatography
Pulsed elution modulation for on-line comprehensive two-dimensional liquid chromatography coupling reversed phase liquid chromatography and hydrophilic interaction chromatography
复制标题
反相液相色谱与亲水相互作用色谱联用的在线全二维液相色谱脉冲洗脱调制
DOI：
10.1016/j.chroma.2018.11.023
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发表时间：
2019-01-04
影响因子：
4.1
通讯作者：
Chen, Bo
中科院分区：
文献类型：
--
作者：
Chen, Yingzhuang;Wu, Yajun;Chen, Bo
本文建立了反相液相色谱(RPLC) -亲水相互作用色谱（HILIC）耦合体系，结合脉冲洗脱反相液相色谱（PE-RPLC）和HILIC对富含皂苷的人参根提取物进行综合分析。通过应用脉冲洗脱（PE）调制技术，所提出的RPLC x HILIC系统可以在两个维度上独立优化色谱分离。在第一个维度（D-1）中，PE调制是通过PE梯度分离复杂混合物来实现的，例如人参根提取物。这个PE梯度包含一组脉冲，其中溶剂强度逐渐增加。因此，D-1洗脱液的调制是通过逐步脉冲分馏来实现的，而不是通过传统的二维界面。此外，馏分数和分馏周期可以独立调节，从而在不损失D-1分离效率的情况下，实现了二维（D-2）分离周期的独立调节。为了克服RPLC x HILIC固有的溶剂不相容性，我们引入了一种新开发的带有旁路的捕获界面。结果表明，人参根提取物中皂苷的分离效果良好。与传统调制方法相比，所提出的RPLC x HILIC系统具有极大的灵活性，可以在大范围内调节调制时间，而无需重新优化D-1 PE梯度。值得一提的是，所提出的RPLC x HILIC系统具有良好的正交性，与传统的基于值的调制方法相比，电流方法可以获得多20%的峰值。两个维度的独立调节可以使所提出的调制方法广泛应用于普通实验室的复杂样品分析。(C) 2018 Elsevier B.V.版权所有
In this work, a reversed phase liquid chromatography (RPLC) coupling to hydrophilic interaction chromatography (HILIC) system has been constructed, combining with pulsed elution reversed phase liquid chromatography (PE-RPLC) and HILIC to comprehensively analyze P. ginseng root extract, which is rich in saponins. By the application of pulsed elution (PE) modulation technique, the proposed RPLC x HILIC system allows the chromatographic separation to be optimized independently in both dimensions. In the first dimension (D-1), PE modulation is achieved by the separation of a complex mixture, such as P. ginseng root extract, with a PE gradient. This PE gradient contains a set of pulses where the solvent strength increases gradually. Thus, the modulation of D-1 eluent is realized by stepwise-pulse fractionation, rather than by a traditional two-dimensional interface. Furthermore, the number of fractions and the fractionated period can be regulated independently, which leads to independent adjustment of the separation cycle in the second dimension (D-2) separation without the loss of D-1 separation efficiency. To overcome the inherent solvent incompatibility of RPLC x HILIC, we introduced a newly developed trapping interface, equipped with bypass. The result indicates excellent separation of saponins in P. ginseng root extract. Compared with the traditional modulation method, the proposed RPLC x HILIC system has extreme flexibility, those modulation time could be regulated in a large range without re-optimizing the D-1 PE gradient. Worthily mentioned, the proposed RPLC x HILIC system shows excellent orthogonality, and 20% more peaks could be obtained with current method compared to the traditional value based modulation method. Independent regulation of both dimensions could enable the proposed modulation method to be widely applied for complex samples analysis in ordinary laboratory. (C) 2018 Elsevier B.V. All rights reserved.