高效凝胶过滤色谱填料KH-s-GFC300的合成与评价

以碱溶法制备的高孔隙度硅胶为基质，与γ-环氧丙基丙氧基三甲氧基硅烷配基进行共价键合，经过酸性水解，使其转化为二醇基，制成了高效亲水凝胶色谱填料。键合反应中采用70 ℃回流冷凝管，使生成的甲醇可顺利地排到体系外，促使反应向有利于键合的方向进行，以获得较高的配基密度。研究表明，水解反应条件与配基密度以及柱效密切相关。当配基密度为2.6~3.5 μmol/m2时，才能显示出较高的柱效。蛋白质分离实验结果显示，此填料的相对分子质量排阻极限为300000，牛血清白蛋白的回收率为99％。

Synthesis and Evaluation of High Performance Gel Filtration Chromatography Packing of KH-s-GFC300

A novel packing for high performance gel filtration chromatography (GFC) was synthesized and characterized. High porosity silica prepared by base-dissolving method was used as the matrix. gamma-(2,3-Epoxy propoxyl) propyltrimethoxysilane was used as the ligand and covalently bonded onto the silica matrix. After acidic hydrolysis, the epoxy groups were converted to the diol groups. Because a condensation tube filled with water at 70 degrees C was used in the grafting reaction, the resulting methanol could easily be discharged from the reaction system to shift the reaction equilibrium to achieve high ligand density. The hydrolysis condition greatly affects ligand density and column efficiency. The high column efficiency is observed when the ligand density is between 2.6 and 3.5 +/- mol/m2. Several proteins, such as cytochrome C, chymotrypsin, ovalbumin, bovine serum albumin, aldolase, ferritin, insulin, gamma-globulin, phosphorylase, actin, carbonic anhydrase, were used to characterize the separation properties of the resulting high performance GFC column. It was shown that the excluded limit of relative molecular mass for the separation of bio-molecules was 300 000. The recovery yield of bovine serum albumin was 99%.