Complete monosaccharide analysis by high-performance anion-exchange chromatography with pulsed amperometric detection

Area of Science:

• Carbohydrate Chemistry
• Analytical Chemistry

Background:

• Monosaccharide analysis is crucial for characterizing complex carbohydrates.
• Existing methods for neutral and amino sugars are established, but acidic sugar analysis is limited.
• Acidic sugars, like uronic and sialic acids, are vital components in many biological glycans.

Purpose of the Study:

• To develop a robust method for analyzing acidic sugars.
• To establish a comprehensive monosaccharide profiling technique for diverse complex carbohydrates.
• To provide a convenient and broadly applicable analytical solution.

Main Methods:

• High-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was optimized for acidic sugar analysis.
• A comprehensive method was developed to profile neutral, amino, and acidic monosaccharides simultaneously.
• The procedure avoids additional derivatization steps, simplifying sample preparation.

Main Results:

• The method demonstrates a low limit of quantitation (~12.5 × 10(-3) nmol) and excellent linearity.
• It was successfully applied to profile monosaccharide compositions of various complex carbohydrates, including glycosaminoglycans, alginate, and fucoidan.
• Hydrolysis patterns of these complex carbohydrates were also investigated.

Conclusions:

• A novel, convenient, and sensitive HPAEC-PAD method enables comprehensive monosaccharide analysis, including rare acidic sugars.
• This technique significantly advances the characterization of complex carbohydrates with diverse structures.
• The method offers broad applicability in glycobiology and related fields.