Journal
GLYCOCONJUGATE JOURNAL
Volume 35, Issue 6, Pages 499-509Publisher
SPRINGER
DOI: 10.1007/s10719-018-9848-1
Keywords
Glycans; Glycomics; Mass spectrometry; Nano-electrospray; Nano-liquid chromatography; Negative ion mode; Orbitrap; Porous graphitized carbon; Post-column make-up flow; Stability
Categories
Funding
- European Commission (EC) [721815, 324400]
- German Research Foundation (DFG) [FOR2509]
- German Federal Ministry of Education and Research (BMBF) [031C557]
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Analysis of glycans via a porous graphitized carbon liquid chromatography (PGC-LC) coupled with electrospray ionization (tandem) mass spectrometry (ESI-MS(/MS)) is a powerful analytical method in the field of glycomics. Isobaric glycan structures can be identified reliably with the help of PGC-LC separation and subsequent identification by ESI-MS(/MS) in negative ion mode. In an effort to adapt PGC-LC-ESI-MS(/MS) to the nano-scale operation, spray instability along the nano-PGC-LC gradient was repeatedly observed on an LTQ Orbitrap Elite mass spectrometer equipped with a standard nano-electrospray ionization source. A stable electrospray was achieved with the implementation of a post-column make-up flow (PCMF). Thereby, acetonitrile was used to supplement the eluate from the nano-PGC-LC column. The improved spray stability enhanced detection and resolution of glycans during the analysis. This was in particular the case for smaller O-glycans which elute early in the high aqueous content regime of the nano-PGC-LC elution gradient. This study introduces PCMF as an easy-to-use instrumental adaptation to significantly improve spray stability in negative ion mode nano-PGC-LC-ESI-MS(/MS)-based analysis of glycans.
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