Simultaneous determination of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan glycosaminoglycan disaccharides by high-performance liquid chromatography using a reverse-phase column with adamantyl groups

Hanamatsu, Hisatoshi; Makino, Satoshi; Ohara, Masatsugu; Suda, Goki; Yokota, Ikuko; Nishihara, Shoko; Sakamoto, Naoya; Furukawa, Yoshihiro

doi:10.1016/j.chroma.2022.463748

Cited by 2 publications

(2 citation statements)

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“…Previous studies have typically used 2-aminobenzamide (2-AB) or 2-aminoacridone (AMAC) as pre-labeling reagents, and 2-cyanoacetamide as a post-labeling reagent to detect the disaccharides using HPLC. [2][3][4][5][6] While obviating the need for a derivatization step, the post-labeling approach generally requires a more complicated HPLC system configuration that incorporates a reaction bath and mixing and cooling coils.…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, removing excess reagents before analysis by HPLC or LC-MS is an essential step, and methods such as gel permeation chromatography using a G-10 column or solid phase extraction techniques are typically used to achieve this aim. 2,3,7,8) However, despite their effectiveness, these steps are time consuming. In a previous study on the analysis of neutral sugars, it was reported that excess reagents could be removed using a commercially available MonoSpin NH 2 column.…”

Section: Introductionmentioning

confidence: 99%

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Development of an Analytical Method for Unsaturated Disaccharides from Heparan Sulfate Using Dansylhydrazine Derivatization and High-Performance Liquid Chromatography with Fluorescence Detection

Ishii,

Aijima,

Yokota

et al. 2024

Biological & Pharmaceutical Bulletin

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Glycosaminoglycans (GAGs), such as heparan sulfate (HS), play essential roles in living organisms. Understanding the functionality of HS and its involvement in disease progression necessitates the sensitive and quantitative detection of HS-derived unsaturated disaccharides. Conventionally, fluorescence derivatization precedes the HPLC analysis of these disaccharides. However, the presence of excess unreacted derivatization reagents can inhibit rapid and sensitive analysis in chromatographic determinations. In this study, we describe analytical methods that use dansylhydrazine as a derivatization agent for the detection and determination of HS-derived unsaturated disaccharides using HPLC. In addition, we have developed a straightforward method for removing excess unreacted reagent using a MonoSpin NH 2 column. This method may be employed to remove excess pre-labeling reagents, thereby facilitating the analysis of HS-derived unsaturated disaccharides with satisfactory reproducibility.

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