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Two-dimensional Liquid Phase Separations Coupled with Mass Spectrometry
Authors: Česla Petr | Vaňková Nikola | Váňová Jana | Nováková Zuzana
Year: 2016
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Two-dimensional Liquid Phase Separations Coupled with Mass Spectrometry The2D separations, which provide increased peak capacity with respect to the unidimensional methods, are becoming essential for separation of complex samples, especially of biological origin. Theoretically, peak capacity of 2D method is given by product of peak capacities achievable in both dimensions based on orthogonal separation principles. In practice, the correlation of separation mechanisms together with undersampling of D1 separation and compatibility of transferred fractions with D2 method decrease the separation power of overall method. Besides proper optimization of separation conditions in both, pressure-driven and electro-driven 2D methods, the increased number of compounds unequivocally identified in the samples can be achieved by utilizing of dual separation system with alternating separation modes (e.g. hydrophilic interaction chromatography with reversed-phase chromatography), or by coupling of 2D separation with MS. In this work, we have investigated possibilities of coupling of 2D LC and LC in combination with CE to MS for separation of glycomic samples. The LC separations utilized fast gradients in HILIC and RP, optimized with the aim of providing improved efficiency with respect to the compression of separated zones. For optimization of gradient conditions, retention times were predicted based on mixed-mode retention model taking into account contribution of adsorption and partitioning of polar molecules into water-rich layer, presented on a surface of stationary phase. The electrodriven separations, employed in 2D setup, were on-line coupled using microfluidic flow-gating interface. Conditions for in-capillary labeling of the samples yielding both, suitable migration characteristics and detection sensitivity of the compounds were tested. The protocols for processing of experimental data from two-dimensional system were developed and verified using pressure-driven and electrodriven separations coupled to mass spectrometric detection.

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