Hydrophilic interaction liquid chromatography in the separation of glycopeptides and their isomers.

摘要: The analysis of intact glycopeptides is a challenge because the structural variety complex conjugates. In this work, we used separation involving hydrophilic interaction liquid chromatography using superficially porous particle HALO® penta-HILIC column with tandem mass spectrometric detection for N-glycopeptides hemopexin. We tested effect mobile phase composition on retention and glycopeptides. results indicated that was combination partitioning adsorption processes. Under optimized conditions, our HILIC method showed ability to efficiently separate glycoforms same peptide backbone including isobaric glycoforms. achieved efficient core outer arm linked fucose bi-antennary tri-antennary SWPAVGNCSSALR glycoform ALPQPQNVTSLLGCTH peptide, respectively. Moreover, demonstrated antennary position sialic acid via α2-6 linkage monosialylated Glycopeptide isomers are often differentially associated various biological Therefore, chromatographic species without need an extensive sample preparation appears attractive their identification, characterization, reliable quantification.