Establishment, Evaluation And Application Of Novel Two-dimensional Liquid Chromatography

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.