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Programmed Fraction Transfer Conditions – Towards More Efficient Focusing in 2D LC

Authors: Česla Petr | Hájek Tomáš | Váňová Jana | Kubát Miroslav | Roušar Tomáš

Year: 2023

Type of publication: ostatní - přednáška nebo poster

Page from-to: nestránkováno

Titles:

Language	Name	Abstract	Keywords
eng	Programmed Fraction Transfer Conditions – Towards More Efficient Focusing in 2D LC	Samples of biological materials are typically highly complex mixtures containing chemical substances with diverse structures and functional groups. The application of two-dimensional chromatography, especially in the comprehensive mode, offers an increased number of peaks, which can be theoretically analyzed and quantified in a single run. One of the significant drawbacks of comprehensive two-dimensional LC, which, up to date, hindered its broader application for real-world samples, is higher detection and quantification limits compared with regular LC due to the excessive dilution of the sample passing through two separation dimensions. Band broadening due to the fraction transfer process can be minimized using short trapping columns instead of sampling loops within the interfacing valve [1]. Several approaches of focusing interfaces in 2D LC were introduced yielding lower dilution of the samples, including fixed and active solvent modulation, and stationary phase-assisted modulation. The transferred analytes are typically focused under steady conditions, which do not necessarily match the optimal conditions for efficien focusing of the whole mixture to be separated. The efficiency of focusing can be significantly affected by applying the tailored gradient conditions, especially in the second dimension. In the setups mentioned above, uniform conditions are typically used during whole 2D analysis so that the focusing efficiency can differ within the gradient profiles used in the first dimension. In the present work, an approach for optimizing programmed conditions in the fraction transfer process between the separation dimensions in LCxLC has been developed. We have used both RPxRP and HILICxRP setups with monolithic 2D LC consisting of polymethacrylate column with zwitterionic sulfobetain functionality [2] in the first dimension combined with silica gel monolithic column in the second dimension for the demonstration of focusing efficiency and compatibility of the m	Two-dimensional liquid chromatography; Focusing interfaces; Gradient elution; Peak compression