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Preparation of a novel polyhydroxy silica stationary phase and application in the separation of alginate oligosaccharides
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Ruiwen JIAO1, Dandan ZHANG1, Xingyuan HU1, Wanting ZHU1, Xiang LI1, 2, 3, Yidi CAI1, 2, 3, Dandan REN1, 2, 3, Qiukuan WANG1, 2, 3, Long WU1, 2, 3, *, Hui ZHOU1, 2, 3, *
Chinese Journal of Chromatography | 2026, 44(4) : 373 - 382
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Chinese Journal of Chromatography | 2026, 44(4): 373-382
• Article •
Preparation of a novel polyhydroxy silica stationary phase and application in the separation of alginate oligosaccharides
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Ruiwen JIAO1, Dandan ZHANG1, Xingyuan HU1, Wanting ZHU1, Xiang LI1, 2, 3, Yidi CAI1, 2, 3, Dandan REN1, 2, 3, Qiukuan WANG1, 2, 3, Long WU1, 2, 3, *, Hui ZHOU1, 2, 3, *
Affiliations
  • 1College of Food Science & Engineering, Dalian Ocean University, Dalian 116023, China
  • 2National R & D Branch Center for Seaweed Processing, Dalian 116023, China
  • 3Key Laboratory of Aquatic Product Processing and Utilization of Liaoning Province, Dalian 116023, China
Published: 2026-04-08 doi: 10.3724/SP.J.1123.2025.03006
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Hyperbranched polyglycerols (HPG) represent an emerging class of dendritic polyethers characterized by a highly branched, three-dimensional architecture and a multitude of terminal hydroxyl groups. This distinctive structural configuration confers exceptional hydrophilicity, biocompatibility, and a high density of modifiable surface functionalities, thus establishing HPG as highly promising materials for the development of advanced chromatographic stationary phases. Their application is particularly relevant for the separation of polar and hydrophilic analytes, which has long posed a significant challenge in conventional reversed-phase liquid chromatography. The integration of such hyperbranched polymers with robust inorganic substrates, such as silica gel, has recently gained traction as a sophisticated materials strategy. This approach synergistically combines the superior mechanical strength and pressure resistance of the inorganic matrix with the rich surface chemistry and tunable hydrophilicity of the polymer, thereby addressing critical limitations of traditional stationary phases in hydrophilic interaction liquid chromatography (HILIC) applications.In the present study, a novel HILIC stationary phase, designated HPG-Sil 3, was synthesized through the in-situ ring-opening polymerization of glycidol monomers from the surface of aminopropyl-functionalized silica microparticles. The synthesis commenced with a meticulous silanization step to graft (3-aminopropyl)triethoxysilane (APTES) onto the silica surface, thereby introducing a uniform layer of primary amine groups. These amine functionalities served as initiation sites for the subsequent grafting-from polymerization, ensuring the formation of a covalently anchored, robust HPG layer. This covalent immobilization strategy is critical for mitigating stationary phase degradation and polymer leaching under prolonged chromatographic use, thereby guaranteeing long-term operational stability. The successful fabrication of the HPG-Sil 3 material and its physicochemical properties were thoroughly characterized using a suite of analytical techniques. Elemental analysis indicated a substantial increase in carbon and hydrogen content post-modification, providing quantitative evidence of organic polymer grafting. Fourier-transform infrared (FTIR) spectroscopy further corroborated this result, revealing signature absorption bands associated with the stretching vibrations of O-H and C-O-C ether linkages, which are characteristic of the HPG polyether structure. Thermogravimetric analysis (TGA) demonstrated the material’s excellent thermal resilience, with the onset of HPG decomposition occurring above 200 ℃, a temperature window far exceeding the operational range of typical HILIC analyses. Textural properties, evaluated via nitrogen physisorption, showed a predictable decrease in specific surface area and pore volume relative to the unmodified silica substrate. This reduction is attributed to the partial filling of the mesoporous silica network by the grafted HPG chains. Importantly, the material retained a sufficiently open porous structure to facilitate efficient mass transfer of analytes during chromatographic runs. To systematically investigate the chromatographic behavior and retention mechanism of the HPG-Sil 3 phase, a set of model polar compounds, including thymine, uracil, hypoxanthine, and adenosine, was selected. The influence of critical mobile phase parameters on the analyte retention factor (k) was examined, including the acetonitrile (ACN) content, the concentration of ammonium acetate buffer, and the buffer pH. The observed retention trends were unequivocally indicative of a dominant HILIC mechanism. A pronounced increase in retention with increasing ACN content was observed for all analytes, consistent with the enhanced partitioning of solutes into a water-rich layer immobilized on the hydrophilic stationary phase surface. Conversely, an increase in buffer concentration led to a decrease in retention, a phenomenon explained by the competitive adsorption of buffer ions with the analytes for polar interaction sites on the HPG layer. Furthermore, the retention of ionizable analytes, namely hypoxanthine and adenosine, was demonstrably influenced by the buffer pH, as pH variations alter their ionization state and thus their overall hydrophilicity and interaction strength with the stationary phase. Under the optimized chromatographic conditions, all four model analytes were baseline separated within a 10-min runtime, exhibiting excellent peak symmetry and high repeatability. The practical utility of the HPG-Sil 3 stationary phase was further demonstrated through the efficient separation of alginate oligosaccharide homologs with degrees of polymerization (DP) ranging from 2 to 7. The elution order followed increasing DP, which aligns perfectly with the HILIC retention principle, as larger oligosaccharides possess more hydroxyl groups and exhibit stronger hydrophilic interactions. The stationary phase also demonstrated remarkable operational stability, with no observable changes in retention times or chromatographic efficiency over 10 consecutive injections, underscoring its robustness for routine analytical applications. In conclusion, the HPG-Sil 3 stationary phase synthesized in this work exhibits outstanding hydrophilic separation performance, a well-understood HILIC retention mechanism, and excellent long-term stability. These attributes position it as a highly competitive and promising material for the analysis of a wide array of polar compounds, including nucleosides, carbohydrates, and polar pharmaceuticals, across diverse fields such as metabolomics, biopharmaceutical analysis, and food chemistry. Future research will focus on the precise control of polymer parameters, including molecular weight, branching density, and post-functionalization, to fine-tune the properties of the HPG layer for tailored selectivity towards specific applications. Furthermore, a systematic evaluation of its performance using complex biological and environmental matrices will be conducted to assess its practical applicability.

hyperbranched polyglycerol  /  hydrophilic chromatographic stationary phase  /  hydrophilic mechanism  /  alginate oligosaccharides
矫芮文, 张丹丹, 胡兴媛, 朱婉婷, 李想, 蔡一镝, 任丹丹, 汪秋宽, 武龙, 周慧. 新型多羟基硅胶固定相的制备及其对褐藻胶寡糖的分离应用. 色谱, 2026 , 44 (4) : 373 -382 . DOI: 10.3724/SP.J.1123.2025.03006
Ruiwen JIAO, Dandan ZHANG, Xingyuan HU, Wanting ZHU, Xiang LI, Yidi CAI, Dandan REN, Qiukuan WANG, Long WU, Hui ZHOU. Preparation of a novel polyhydroxy silica stationary phase and application in the separation of alginate oligosaccharides[J]. Chinese Journal of Chromatography, 2026 , 44 (4) : 373 -382 . DOI: 10.3724/SP.J.1123.2025.03006
Year 2026 volume 44 Issue 4
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doi: 10.3724/SP.J.1123.2025.03006
  • Receive Date:2025-03-06
  • Online Date:2026-10-27
  • Published:2026-04-08
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  • Received:2025-03-06
Affiliations
    1College of Food Science & Engineering, Dalian Ocean University, Dalian 116023, China
    2National R & D Branch Center for Seaweed Processing, Dalian 116023, China
    3Key Laboratory of Aquatic Product Processing and Utilization of Liaoning Province, Dalian 116023, China
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表12种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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