Abstract:

Objective: To extract glycosaminoglycans (GAGs) from hard clams (Meretrix meretrix) by multi-enzymatic
hydrolysis and to analyze the structure of chondroitin sulfate in the extract by infrared (IR) spectroscopy. Methods: The
components of GAGs were examined by ultra high performance liquid chromatography-electrospray-mass spectrometry
(UPLC-ESI-MS) using an ACQUITY UPLC BEH Amide chromatographic column (2.1 mm × 100 mm, 1.7 μm) and an
ACQUITY UPLC BEH Amide VanGuard guard column (2.1 mm × 5 mm, 1.7 μm). The mobile phase was composed of a
mixture of water and methanol (75:25, V/V) containing 0.2% glacial acetic acid at a flow rate of 0.1 mL/min. The injection
volume was set as 20 μL. Results: Rapid separation of the components was achieved and their structures were determined by
UPLC-ESI-MS. Conclusion: This paper provides an effective method to identify the structure of GAGs.

Key words: hard clams, glycosaminoglycans, chondroitin sulfate, ultra high performance liquid chromatographyelectrospray- mass spectrometry (UPLC-ESI-MS), infrared spectroscopy (IR)