Development of highly efficient and highly selective HPLC columns that provide multiple polar separation mechanisms simultaneously

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K05900
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: Ikegami Tohru 京都工芸繊維大学, 分子化学系, 准教授 (20301252)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 液体クロマトグラフィー / 親水性相互作用クロマトグラフィー / 分離媒体 / 分離特性 / 重合修飾

Outline of Final Research Achievements

The relationship between the optimization of the polymerization reaction used for the surface modification of silica and the separation performance was studied. Poly(acrylamide) type silica was prepared by surface-initiated atom transfer radical polymerization (SI-ATRP) to obtain hydrophilic interaction chromatography (HILIC) columns providing highly retentive and good selectivity for polar biomolecules. The relationship between the degree of polymerization or surface coverage of the polymer and the separation characteristics was explored to achieve high hydrophilicity and great retention simultaneously with silica particles of 20 nm pores.
Using the column developed by us, fine separation of structural isomers of monosaccharides and oligosaccharides found in glycoproteins was carried out (with international joint work). Furthermore, anomeric separation of sugar was also performed, and the equilibrium constant of the process was measured.

Free Research Field

クロマトグラフィー

Academic Significance and Societal Importance of the Research Achievements

本研究で合成した表面開始原子移動ラジカル重合型のポリアクリルアミドカラムは市販されているどのアミド型カラムとも違う特徴を示した。すなわち、シリカ表面に濃縮される水の層が厚く、弱い酸性の表面を提供できる。親水性の保持が大きく、かつ構造の差異に対する選択性が高い固定相であり、実用性が高いと判断される。また、糖タンパクに含まれる糖鎖の研究は、現在注目を集めている抗体医薬の製造や同一性の確認に不可欠であり、社会的意義が大きいと考える。