Determination of N-Glycopeptides by Hydrophilic Interaction Liquid Chromatography and Porous Graphitized Carbon Chromatography with Mass Spectrometry Detection. (22nd January 2017)

• Record Type: Journal Article
• Authors: Lu, Jun; Fu, Dongmei; Yu, Long; Cao, Cuiyan; Zou, Lijuan; Liang, Xinmiao
• Is Part Of: Analytical letters. Volume 50:Number 2(2017)
• Page Start: 315
• Page End: 324
• Publication Date: 2017-01-22
• Subjects: Glycan -- glycopeptide -- hydrophilic interaction chromatography -- mass spectrometry -- porous graphitized carbon; Chemistry, Analytic -- Periodicals; Chemistry, Analytic -- Abstracts; 543
• DOI: 10.1080/00032719.2016.1181644
• Languages: English
• ISSNs: 0003-2719

Abstract

ABSTRACT: An offline two-dimensional chromatographic method based on the combination of hydrophilic interaction liquid chromatography (HILIC) and porous graphitized carbon (PGC) chromatography was developed for the separation and purification of glycopeptides. The high selectivity of HILIC and PGC isolated high-purity isomers of N -glycopeptides from ribonuclease B. N -Glycopeptides were first separated from nonglycosylated peptides, and N -glycopeptides were sorted into fractions through the first-dimensional HILIC according to their monosaccharides. Further separation of the glycopeptide isomers in each fraction was achieved using second-dimensional PGC. Structural differences of the glycopeptide isomers were further enzymatically hydrolyzed with peptide- N -glycosidase F. The glycan structure were elucidated by matrix assisted laser desorption ionization tandem quadrupole time-of-flight mass spectrometry. The established procedure allows the isolation of glycopeptide or glycan standards from natural sources.