title: Online coupling of high-resolution chromatography with extreme UV photon activation tandem mass spectrometry: Application to the structural investigation of complex glycans by dissociative photoionization.
abstract: The activation of ions by extreme-energy photons (XUV) produced by a synchrotron radiation beamline is a powerful method for characterizing complex glycans using tandem mass spectrometry (MS). As previously described, this activation method leads to rich fragmentation spectra with many structurally valuable cross-ring cleavages while maintaining labile modifications on the glycan structures. However, until now, the tandem MS event was too long to be compatible with liquid chromatography elution times. In this work, the duty cycle of the activation and detection of fragments was shortened, and the background signal on the spectra was drastically reduced. Both improvements allowed, for the first time, the successful coupling of a UHPLC system to XUV-activated tandem MS. The approach was used to characterize a complex mixture of oligo-porphyrans, which are a class of highly sulfated oligosaccharides, in a fully automated way. Due to an enhanced dynamic range and an increased sensitivity, some hypothetical structures of low abundance have been unequivocally confirmed in this study and others have been revised. Some previously undescribed species of oligo-porphyrans that exhibit lateral branching have been fully resolved. This work contributes to the scarce knowledge of the structure of porphyrans in red algae and pushes the current capacities of XUV-activation tandem MS by demonstrating the possibility of a direct coupling with UHPLC. This study will considerably broaden the applicability and practicality of this method in many fields of analytical biology.
authors: INRA, UR1268 Biopolymers Interactions Assemblies F-44316 Nantes, France. Electronic address: David.Ropartz@nantes.inra.fr.
authors: David
authors: D
authors: Ropartz
authors: Synchrotron SOLEIL, L'Orme des Merisiers, F-91190 Gif-sur-Yvette, France; UAR 1008 CEPIA, INRA, F-44316 Nantes, France.
authors: Alexandre
authors: A
authors: Giuliani
authors: INRA, UR1268 Biopolymers Interactions Assemblies F-44316 Nantes, France.
authors: Mathieu
authors: M
authors: Fanuel
authors: Sorbonne Universités, Université Pierre et Marie Curie, Paris VI, CNRS, Integrative Biology of Marine Models, UMR 8227, Station Biologique, Place George Teissier, F29688 Roscoff Cedex, France.
authors: Cécile
authors: C
authors: Hervé
authors: Sorbonne Universités, Université Pierre et Marie Curie, Paris VI, CNRS, Integrative Biology of Marine Models, UMR 8227, Station Biologique, Place George Teissier, F29688 Roscoff Cedex, France.
authors: Mirjam
authors: M
authors: Czjzek
authors: INRA, UR1268 Biopolymers Interactions Assemblies F-44316 Nantes, France.
authors: Hélène
authors: H
authors: Rogniaux
mesh_terms: D002851:Chromatography, High Pressure Liquid; D017785:Photons; D011134:Polysaccharides / Q000032:analysis*; D012685:Sepharose / Q000031:analogs & derivatives / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014466:Ultraviolet Rays
chemical_list: D011134:Polysaccharides; C038549:porphyran; D012685:Sepharose
keywords: Direct coupling; Extreme ultra-violet photon activation; High throughput; High-resolution liquid chromatography; Structures of minor oligosaccharides; Tandem mass spectrometry