Hanamatsu Hisatoshi, Makino Satoshi, Ohara Masatsugu, Suda Goki, Yokota Ikuko, Nishihara Shoko, Sakamoto Naoya, Furukawa Jun-Ichi
Department of Gastroenterology and Hepatology, Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido 060-8638, Japan; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido 060-8638, Japan.
Sanyo Fine IRICA Technology Co., Ltd. Kyoto, Kyoto 601-8037, Japan.
J Chromatogr A. 2023 Jan 25;1689:463748. doi: 10.1016/j.chroma.2022.463748. Epub 2022 Dec 23.
Glycosaminoglycans (GAGs), which are one of the major components of proteoglycans, play a pivotal role in physiological processes such as signal transduction, cell adhesion, growth, and differentiation. Characterization of GAGs is challenging due to the tremendous structural diversity of heteropolysaccharides with numerous sulfate or carboxyl groups. In this present study, we examined the analysis of 2-aminobenzamide (2-AB) labeled GAG disaccharides by high-performance liquid chromatography (HPLC) using a reverse-phase (RP)-column with adamantyl groups. Under the analytical conditions, 17 types of 2-AB labeled GAG disaccharides derived from heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan were sequentially separated in a single analysis. The analysis time was fast with high retention time reproducibility. Moreover, the RP-HPLC column with adamantyl groups allowed the quantification of GAGs in various biological samples, such as serum, cultured cells, and culture medium.
糖胺聚糖（GAGs）是蛋白聚糖的主要成分之一，在信号转导、细胞黏附、生长和分化等生理过程中起关键作用。由于具有众多硫酸基或羧基的杂多糖结构多样性巨大，GAGs的表征具有挑战性。在本研究中，我们使用带有金刚烷基的反相（RP）柱，通过高效液相色谱（HPLC）对2-氨基苯甲酰胺（2-AB）标记的GAG二糖进行了分析。在分析条件下，单次分析可依次分离出17种源自硫酸乙酰肝素、硫酸软骨素/硫酸皮肤素和透明质酸的2-AB标记的GAG二糖。分析时间快，保留时间重现性高。此外，带有金刚烷基的RP-HPLC柱可对血清、培养细胞和培养基等各种生物样品中的GAGs进行定量分析。
