A hyperbranched polymer-based stationary phase for hydrophilic interaction chromatography

Area of Science:

• Analytical Chemistry
• Polymer Science
• Chromatography

Background:

• Hydrophilic interaction chromatography (HILIC) is crucial for separating polar analytes.
• Existing HILIC stationary phases often suffer from high column bleed and limited efficiency for diverse polar compounds.
• Development of robust, highly hydrophilic stationary phases with low bleed is essential for advancing chromatographic separations.

Purpose of the Study:

• To synthesize and characterize a novel polymer-based stationary phase for HILIC.
• To evaluate the chromatographic performance, including separation efficiency and retention mechanisms.
• To assess the bleed characteristics of the new stationary phase compared to commercial alternatives.

Main Methods:

• Grafting cysteamine onto hydrolyzed poly(glycidyl methacrylate-divinylbenzene) microspheres.
• One-pot synthesis of hyperbranched poly(thiol ether amine) via sequential thiol-ene and epoxy-amine click reactions.
• Characterization of hydrophilicity and evaluation of separation for vitamins, amino acids, organic acids, sugars, and nucleosides.

Main Results:

• The synthesized stationary phase demonstrated high hydrophilicity and excellent dispersibility in aqueous solutions.
• Efficient separation of a broad range of polar analytes, including vitamins, amino acids, organic acids, and sugars, was achieved.
• The HILIC column exhibited negligible bleed levels (2.1 to 554.8 times lower than commercial columns) under gradient elution with up to 50% aqueous fraction.

Conclusions:

• The developed hyperbranched polymer-based stationary phase is highly effective for HILIC separations of polar compounds.
• The low bleed characteristic significantly improves chromatographic performance and method robustness.
• This novel material offers a promising alternative to existing HILIC stationary phases, particularly for sensitive analyses.