Improvement of Electrospray Stability in Negative Ion Mode for Nano-PGC-LC-MS Glycoanalysis Via Post-column Make-up Flow
Overview
Endocrinology
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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.
Wojtkiewicz M, Subramanian S, Gundry R Anal Chem. 2024; 96(15):5746-5751.
PMID: 38556995 PMC: 11024887. DOI: 10.1021/acs.analchem.3c03649.
Fitz V, Abiead Y, Berger D, Koellensperger G Front Mol Biosci. 2022; 9:857505.
PMID: 35923463 PMC: 9340153. DOI: 10.3389/fmolb.2022.857505.
Young C, Condina M, Briggs M, Moh E, Kaur G, Oehler M Front Chem. 2021; 9:653959.
PMID: 34178940 PMC: 8226321. DOI: 10.3389/fchem.2021.653959.
State-of-the-Art Glycomics Technologies in Glycobiotechnology.
Pralow A, Cajic S, Alagesan K, Kolarich D, Rapp E Adv Biochem Eng Biotechnol. 2020; 175:379-411.
PMID: 33112988 DOI: 10.1007/10_2020_143.
Nano-liquid chromatography-mass spectrometry and recent applications in omics investigations.
Sanders K, Edwards J Anal Methods. 2020; 12(36):4404-4417.
PMID: 32901622 PMC: 7530103. DOI: 10.1039/d0ay01194k.