Comparison of hydrophilic-interaction, reversed-phase and porous graphitic carbon chromatography for glycan analysis

Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species

“…44 For the analysis of larger complex glycans RP separation would be most efficient especially when coupled with mass spectrometry. 9,41,45,46 Such coupling however, increases analysis times and requires further expertise. Table 1 shows the different glucose units using a dextran ladder (GU), arabinose ladder (AU) and maltooligosaccharide ladder (GU CE ) values of various structures identified from IgG glycans in 530 human.…”

Recent advances in hydrophilic interaction liquid chromatography (HILIC) for structural glycomics

“…A recently introduced sub-2-mm amido UPLC HILIC phase has been successfully used for analyzing tagged glycans with fluorescence and mass spectrometric detection (see also Table 1, Section HPLC) [18][19][20][21]. The major advantages of this rather new technology are the increased separation efficiency and the associated resolution accompanied by increased analytical speed leading to higher sample throughput.…”

“…Another very interesting report on the use of the 1.7-mm HILIC phase UPLC BEH glycan column was published recently, in which the performance of the HILIC column online coupled to a linear ion-trap Orbitrap instrument was compared with the separation by ion-pairing reversed-phase and porous graphitized carbon (PGC) chromatography [18]. The 1.7-mm HILIC phase column showed its strength in the analysis of complex glycans samples as a steep gradient could be applied and the availability of the UPLC stationary phase proved superior peak capacity.…”

Liquid Chromatography – Mass Spectrometry of Carbohydrates Derivatized with Biotinamidocaproyl Hydrazide

“…Comparative examinations of pre-column labeled BACH-monosaccharides were performed by means of reversed phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) as these methods shown to be highly complementary in resolution of mixtures of carbohydrates (Melmer et al, 2011;Schlichtherle-Cerny et al, 2003;Strege et al, 2000). HILIC is based on the hydrophilic character of the analyte which accounts for the interaction of the molecule with the stationary phase (Wuhrer et al, 2009).…”

“…Most of the labels used for the derivatization of carbohydrates possess a chromophor or a flurophor, enabling a sensitive detection of these analytes by means of spectroscopic m e t h o d s . A p a r t f r o m a n i n c r e a se in detection sensitivity, chromatographic properties of derivatized sugars may improve in comparison with their underivatized counterparts (Melmer et al, 2011;Pabst et al, 2009). Furthermore, mass spectrometric signal intensity showed to profit from the derivatization step, as an effective ionization was achieved through derivatization.…”