Collective sampling of intact anionic polysaccharide components and application in quantitative determination by...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• Anionic polysaccharides, including glycosaminoglycans (GAGs) and alginate, are challenging to analyze quantitatively.
• Traditional electrospray mass spectrometry (ESI-MS) can cause source-induced fragmentation of these molecules.
• This fragmentation converts intact polysaccharides into smaller, structurally characteristic saccharides.

Purpose of the Study:

• To develop a novel Liquid Chromatography-Mass Spectrometry (LC-MS) method for the quantitative analysis of intact anionic polysaccharides.
• To leverage source-induced fragmentation for collective detection of polysaccharides.
• To demonstrate the feasibility of this approach with a mixture of common anionic polysaccharides.

Main Methods:

• Electrospray mass spectrometry (ESI-MS) with high source cone voltage to induce polysaccharide fragmentation.
• Ion-pairing reverse-phase liquid chromatography (IP-RP-LC) for the separation of polysaccharides into compact bands.
• Detection of characteristic small saccharide fragments for collective polysaccharide identification and quantification.

Main Results:

• Source-induced fragmentation effectively converts anionic polysaccharides into detectable small saccharide fragments.
• IP-RP-LC enables the elution of polysaccharides as distinct, manageable bands.
• The combined LC-MS approach allows for the collective detection and quantitative analysis of intact anionic polysaccharides.

Conclusions:

• A novel LC-MS method has been developed for the quantitative analysis of intact anionic polysaccharides.
• This method utilizes source-induced fragmentation and ion-pairing reverse-phase liquid chromatography.
• The approach is feasible for analyzing mixtures of heparin, chondroitin sulfate A, and alginate.