Exploration of Mesoporous Siliceous Particle-Based High-Performance Size Exclusion Chromatography for the Quantitation of Biomacromolecular Polysaccharides

Abstract

Based on a modification of normal hydrothermal synthesis, uniform and spherical mesoporous siliceous particles (MSP) were prepared and applied to high-performance size exclusion chromatography (HPSEC). Sodium acetate buffer solution at pH = 4.0 was the optimal mobile phase for the separation of the three biomacromolecular polysaccharides (alginic acid, fucoidan, and laminarin) in the MSP-based HPSEC. Based on the optimal separation conditions, the calibration curves of the three polysaccharide standards showed good linearity (R² > 0.999). In addition, the limits of detection of the three standards ranged from 0.02 to 0.13 mg mL⁻¹, the limits of quantitation ranged from 0.07 to 0.43 mg mL⁻¹, the related recoveries were 97.7–101.2%, and the relative standard deviations of the retention time were 0.2–0.5. All the quantitative parameters showed that the developed approach was suitable for quantifying biomacromolecular polysaccharides. Compared to a common commercial HPSEC column, the MSP-based HPSEC column was low cost and had a similar separation efficiency for the three polysaccharides. In particular, 2.8 mg g⁻¹ of the alginic acid fractions, 4.4 mg g⁻¹ of the fucoidan fractions, and 6.9 mg g⁻¹ of the laminarin fractions were separated from the Thallus Laminariae extracts by MSP-based HPSEC. Similar results were obtained from the commercial HPSEC.

Keywords:

• biomacromolecule
• high-performance size exclusion chromatography
• mesoporous siliceous particles
• polysaccharides
• quantitation
• separation