Zoltan Szabo1, Niclas G. Karlsson2, Yury Agroskin1, Jim Thayer1, Gina Tan3, Rosa Viner3, Andreas Huhmer3, Jeff Rohrer1, Liu Yan1, Kannan Srinivasan1, Dietmar Reusch4 and Christopher A. Pohl1 ASMS 2017 Poster In this study, High Performance Anion Exchange Chromatography (HPAE) coupled to an HRAM Orbitrap™ mass spectrometer is demonstrated to be an efficient tool to separate and sequence sulfated O-glycan alditols. Porcine gastric mucin (type III) harbors high levels of sulfated but fewer sialylated glycans. After release and rapid purification, these O-linked glycans were directly injected and analyzed. However, bovine submaxillary mucin harbors fewer sulfated but more sialylated Oglycans. For this sample, a new workflow, based on weak anion exchange (WAX) fractionation, enabled the enrichment of sulfated glycans, minimizing ion-suppression by the sialylated glycans. Using a post column splitter, 60 % of the flow was desalted with an electrochemically regenerated desalter prior to introduction into the mass spectrometer, the balance going to the electrochemical detector. Due to the ability of HPAE to resolve isomers, 27 sulfated O linked-glycans were identified from Porcine gastric mucin (type III), and 9 from bovine submaxillary mucin. Using the data-dependent mode of the Thermo Scientific™ Q Exactive™ hybrid quadrupole-Orbitrap mass spectrometer, highly informative MS2 spectra enabled confident identification of the separated species.
1 ThermoFisher Scientific, Sunnyvale, CA, USA
2 University of Gothenburg, Gothenburg, Sweden
3 ThermoFisher Scientific, San Jose, CA, USA, 4 Roche Diagnostics GmbH, Penzberg, Germany