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Licensed Unlicensed Requires Authentication Published by De Gruyter April 10, 2009

On-line nano-HPLC/ESI QTOF MS monitoring of α2–3 and α2–6 sialylation in granulocyte glycosphingolipidome

  • Stephan Kirsch , Johannes Müthing , Jasna Peter-Katalinić and Laura Bindila
From the journal Biological Chemistry

Abstract

A novel glycosphingolipidomic protocol using nano-high performance liquid chromatography coupled on-line to electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOF-MS) focusing on the separation of isomeric ganglioside structures is described here. A highly efficient separation of α2–3- and α2–6-sialylated ganglioside species of different carbohydrate chain length was achieved on an HILIC-amido column, followed by sensitive flow-through ESI-QTOF-MS detection and unambiguous structural identification by tandem MS experiments. The protocol was applied to encompass the glycosphingolipidome of human granulocytes, where 182 distinct components could be clearly identified and assigned regarding the ganglioside type and the isomer distribution.


Corresponding author

Received: 2009-1-21
Accepted: 2009-3-22
Published Online: 2009-04-10
Published in Print: 2009-07-01

©2009 by Walter de Gruyter Berlin New York

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