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Développement de nouvelles méthodes séparatives compatibles avec une détection par spectrométrie de masse et par électrochimie pour l'analyse des traces de catécholamines et molécules apparentées

Abstract : As neurotransmitters, catecholamines play an important role in the control and regulation of numerous brain functions. They are also believed to be implicated in different neurodegenerative disorders. First an ion pairing chromatography method using nonafluoropentanoic acid as volatile ion paring agent was developed on the new generation of C18 columns (monolith and fused core). This method is compatible with MS detection in positive ionization mode. Secondly an HILIC method was optimized using different commercially available HILIC supports, they can be classified as follows: neutral (diol, amide, and cyano bounded), positively charged (amino, triazole bounded), negatively charged (bare silica as wholly porous particles or fused core particles columns) and zwitterionic (sulfobetaine bounded). Our studies lead us to a better understanding of the HILIC retention mechanism and also to the selection of the most appropriated column for catecholamine analysis. Only the HILIC system was compatible with both positive and negative ionization modes. The two chromatographic systems were then compared in terms of resolution, efficiency, detection and quantification limits (LOD/LOQ), calibration linearity and repeatability. The LODs obtained were in the range of 1-100 ng.mL-1. A simple pre-concentration method using Oasis HLB and PGC solid phase extraction cartridges has been optimized in order to enhance the LODs. Finally the optimized SPE-LC-MS/MS method has been applied to the identification of these compounds present in brain extracts.
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Submitted on : Friday, June 4, 2010 - 10:46:20 AM
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  • HAL Id : tel-00489172, version 1

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Raluca-Ioana Chirita. Développement de nouvelles méthodes séparatives compatibles avec une détection par spectrométrie de masse et par électrochimie pour l'analyse des traces de catécholamines et molécules apparentées. Autre. Université d'Orléans, 2009. Français. ⟨tel-00489172⟩

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