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On-line nano-HPLC/ESI QTOF MS monitoring of alpha2-3 and alpha2-6 sialylation in granulocyte glycosphingolipidome.

Stephan S Kirsch, Johannes J Müthing, Jasna J Peter-Katalinić, and Laura L Bindila

Biol Chem 390(7):657-72 (2009) PMID 19361287

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 alpha2-3- and alpha2-6-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.

DOI: 10.1515/BC.2009.066
Version: za2963e q8za9 q8zb8 q8zc2 q8zd5 q8ze4 q8zf6 q8zgb

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