On-line nano-HPLC/ESI QTOF MS monitoring of alpha2-3 and alpha2-6 sialylation in granulocyte glycosphingolipidome.
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.
Kirsch
Stephan
S
Institute of Medical Physics and Biophysics, University of Münster, D-48149 Münster, Germany.
Müthing
Johannes
J
Peter-Katalinić
Jasna
J
Bindila
Laura
L
D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D005732:Gangliosides / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D006098:Granulocytes / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D036103:Nanotechnology; D021241:Spectrometry, Mass, Electrospray Ionization
D005732:Gangliosides