新型二维液相色谱体系的建立、评价及其应用
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摘要
本论文针对二维液相色谱发展中所面临的问题,从复杂样品分析的需求出发,展开了二维液相色谱体系的理论研究和实际应用。结合本课题组开发的新型分离材料,建立了基于联萘柱与C18柱的二维反相／反相液相色谱系统,以中心切割方式在第一维选择性富集大黄中的蒽醌,在第二维以液质联用方法对大黄中的蒽醌进行分离和鉴别。
建立了基于环糊精柱和C18柱的二维亲水／反相液相色谱系统,以蛹虫草为研究对象,优化了第一维的亲水色谱条件,对第二维反相色谱分离中pH值对分离选择性进行考察,最终确定了pH 2.8为优化条件。采用离线二维的方式,在第一维将蛹虫草组分划分为12个组分,第二维对组分的分离可以将第一维中共流出的组分进行了有效的分离。亲水-反相二维系统具有较好的正交性,利用该方法可以发现复杂体系中更多的化学物质基础,特别是微量组分。
捕获柱是重要的二维接口。通过定量地比较第一维色谱柱接捕获柱和不接捕获柱的保留时间,峰面积和峰展宽差异,建立了捕获柱的评价体系,考察了影响捕获柱捕获能力的主要因素。通过该方法可以对捕获柱进行筛选,考察样品含量及性质对捕获能力的影响,直观比较和优化对复杂样品的捕获能力。
In this thesis, the theoretical study and practical application of two-dimensional liquid chromatography (2D-LC) were developed to get better analysis of complicated samples. With the introduction of novel stationary phases, we established the heart-cutting mode 2D-RPLC/RPLC by "click" binaphthyl column and C18 column, selectively enriching anthraquinones from Rheum palmatum L. in the first dimension, then separating and identifying anthraquinones by UPLC-ESI-MS/MS in the second dimension.
Take Cordyceps militaris(L.ex Fr.)Link as research object, a 2D-HILIC/RPLC was developed basing on "click"β-CD column and C18 column. The hydrophilic chromatographic conditions were optimized in the first dimension and the selectivity affected by pH values was investigated in the second dimension, ultimately determined the pH 2.8 as the optimal condition. In the off-line 2D-LC, the Cordyceps militaris(L.ex Fr.)Link was divided into 12 fractions in the first dimension and then all the fracions were separated in the second dimension. The experiment results indicated that the orthogonality of the 2DLC system is good, and this kind of method can be used to discover more chemical components in the complex samples, especially trace quantities.
Capture column is one of most important two-dimensional interfaces. An evaluation system of capture column was established by quantative comprison of retention times, peak areas and peak spreads through connecting and inconnecting the capture columns. We have reviewed the main factors which impacted the capture columns'abilities, selected the capture columns by this method and researched the influence of injection volumes and properties towards the columns'capture abilities, finally complex samples'capture abilities were compared and optimized directly by this method.
建立了基于环糊精柱和C18柱的二维亲水／反相液相色谱系统,以蛹虫草为研究对象,优化了第一维的亲水色谱条件,对第二维反相色谱分离中pH值对分离选择性进行考察,最终确定了pH 2.8为优化条件。采用离线二维的方式,在第一维将蛹虫草组分划分为12个组分,第二维对组分的分离可以将第一维中共流出的组分进行了有效的分离。亲水-反相二维系统具有较好的正交性,利用该方法可以发现复杂体系中更多的化学物质基础,特别是微量组分。
捕获柱是重要的二维接口。通过定量地比较第一维色谱柱接捕获柱和不接捕获柱的保留时间,峰面积和峰展宽差异,建立了捕获柱的评价体系,考察了影响捕获柱捕获能力的主要因素。通过该方法可以对捕获柱进行筛选,考察样品含量及性质对捕获能力的影响,直观比较和优化对复杂样品的捕获能力。
In this thesis, the theoretical study and practical application of two-dimensional liquid chromatography (2D-LC) were developed to get better analysis of complicated samples. With the introduction of novel stationary phases, we established the heart-cutting mode 2D-RPLC/RPLC by "click" binaphthyl column and C18 column, selectively enriching anthraquinones from Rheum palmatum L. in the first dimension, then separating and identifying anthraquinones by UPLC-ESI-MS/MS in the second dimension.
Take Cordyceps militaris(L.ex Fr.)Link as research object, a 2D-HILIC/RPLC was developed basing on "click"β-CD column and C18 column. The hydrophilic chromatographic conditions were optimized in the first dimension and the selectivity affected by pH values was investigated in the second dimension, ultimately determined the pH 2.8 as the optimal condition. In the off-line 2D-LC, the Cordyceps militaris(L.ex Fr.)Link was divided into 12 fractions in the first dimension and then all the fracions were separated in the second dimension. The experiment results indicated that the orthogonality of the 2DLC system is good, and this kind of method can be used to discover more chemical components in the complex samples, especially trace quantities.
Capture column is one of most important two-dimensional interfaces. An evaluation system of capture column was established by quantative comprison of retention times, peak areas and peak spreads through connecting and inconnecting the capture columns. We have reviewed the main factors which impacted the capture columns'abilities, selected the capture columns by this method and researched the influence of injection volumes and properties towards the columns'capture abilities, finally complex samples'capture abilities were compared and optimized directly by this method.
引文
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