Advancements in glycan analysis: high performance anion exchange chromatography-pulsed amprometric detection coupled with mass spectrometry for structural elucidation.

Carbohydrates are essential biomolecules that play a vital role in various biological processes across humans, plants, and bacteria. Despite their ubiquity, the structural elucidation of carbohydrates, particularly oligo- and polysaccharides, remains a significant challenge due to their complex and heterogeneous nature. The high-performance anion exchange chromatography (HPAEC) or called ion chromatography (IC) coupled with pulsed amperometric detection (PAD) has emerged as a powerful tool for highly effective separation and highly specific detection of glycans. The introduction of mass spectrometry (MS) into HPAEC-PAD systems has further advanced glycan analysis by enabling detailed structural elucidation, including branching, linkage patterns, and sequence determination. The use of suppressor technology allows for the coupling of HPAEC with MS by converting non-volatile salts in the mobile phase into volatile ones. This review highlights the current advancements in HPAEC-PAD/MS for oligo- and polysaccharide structural analysis, discussing the strengths and limitations of different suppressor systems, the role of MS in glycan analysis, and the emerging applications of this technology in the field of glycomics. With continued innovation, HPAEC-PAD/MS is poised to become an essential tool for the detailed characterization of polysaccharides, supporting advancements in pharmaceutical, biomedical, and biotechnological research.