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Optimisation de méthodes bidimensionnelles en ligne LCxLC-UV/MS et LCxSFC-UV pour l’analyse d’échantillons complexes

Abstract : Comprehensive two-dimensional liquid chromatography is a powerful but complex separative technique. After detailing the interest of such a technique, the method development issues and the experimental conditions employed throughout this work, a particular attention is paid to the optimization of LCxLC methods. Accordingly an optimization procedure based on Pareto-optimal method is described. The predicted optimal conditions are then applied to experimental RPLCxRPLC separations of complex samples of peptides and compared with 1D-RPLC in terms of peak capacity, analysis time and sensitivity clearly showing the advantage of RPLCxRPLC approach.The optimization of a HICxRPLC-UV/MS method for the exhaustive characterization of an antibody-drug conjugate is achieved highlighting the high complementarity of the different detection modes used both in 1D and 2D. Finally, a proof of concept concerning the implementation of RPLCxSFC coupling is achieved with the aim of increasing the separation space coverage for neutral compounds. The optimized RPLCxSFC separation is then compared with an optimized RPLCxRPLC approach for the analysis of a bio-oil sample showing that RPLCxSFC is a credible alternative for the separation of such a sample
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Morgan Sarrut. Optimisation de méthodes bidimensionnelles en ligne LCxLC-UV/MS et LCxSFC-UV pour l’analyse d’échantillons complexes. Chimie analytique. Université de Lyon, 2016. Français. ⟨NNT : 2016LYSE1189⟩. ⟨tel-01468223⟩

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