Two-dimensional hydrophilic interaction chromatography coupling anion-exchange and hydrophilic interaction columns for separation of 2-pyridylamino derivatives of neutral and sialylated N-glycans

Glycan labeling strategies and their use in identification and quantification

“…An extensive database has been developed which uses the standardized elution positions of 2-AB-labeled glycans in hydrophilic interaction liquid chromatography (HILIC) with fluorescence detection for structural assignment [29]. PA is also widely used in high performance liquid chromatography (HPLC) profiling [30, 31], and databases for structural assignments based on standardized elution positions have been developed as well [32]. A major drawback of PA is the necessary recrystallization step prior to its use, since the commercially available compound lacks sufficient purity.…”

Separation of 2-aminobenzamide labeled glycans using hydrophilic interaction chromatography columns packed with 1.7μm sorbent

“…The most common approaches combine HILIC and RP-LC separation modes [7], or HILIC and HPAEC methods [14,15], where anion-exchange chromatography is utilized to fractionate glycans according to their charge or number of sialic acid residues in the glycan structure. Combinations of all three LC separation modes with normalized retention time GU plots have been reported [7].…”

Theory and Practice of UHPLC and UHPLC–MS

“…The resolution was the best at pH 6.8, but the orthogonality against a C 18 phase was better at pH ¼ 3 (Boersema et al, 2007). HILIC on a capillary zwitterionic ZIC-cHILIC column in combination with electrospray ionization mass spectrometry has been applied to the detection and identification of more than 100 N-glycopeptides and O-glycopeptides in a single run (Takegawa et al, 2008). Glycopeptides could be distinguished by their glycan composition on a monolithic silica gel capillary surface modified with sulfobetaine stationary phase, in contrast to RP-HPLC (Wohlgemuth et al, 2010).…”