乙二醇-磷霉素改性氧化锆色谱固定相的制备及其在蛋白质分离中的应用

Preparation of Glycol-Fosfomycin Modified Zirconia Chromatography Stationary Phases and Its Application to the Separation of Proteins

合成了分离蛋白质的乙二醇-磷霉素钠改性氧化锆高效液相色谱固定相,通过漫反射红外光谱、元素分析等分析方法对该固定相进行了表征.以溶菌酶、核糖核酸酶A、细胞色素C和糜蛋白酶四种标准碱性蛋白质为探针,系统地考察了固定相的疏水相互作用色谱性能.结果表明,乙二醇-磷霉素改性氧化锆固定相对蛋白质有一定的保留,表现出较高的分离选择性.

A novel glycol-fosfomycin modified zirconia （G-F-ZrO2） stationary phase was prepared and characterized by elemental analysis, FT-IR and N2 adsorption. Its chromatographic performance was evaluated using four standard proteins including lysozyme （Lys）, ribonuclease-A （RNase-A）, a-chymotrypsin （a-Chy） and Cytochrome C （Cyt-C） as probes, and the effects of salt concentration, pH, salt type in eluents and temperature on retention behavior of proteins were investigated. The results indicate that the stationary phase behaves mainly as a hydrophobic interaction chromatographic packing with salt concentration in the mobile phase above 1.0 mol·L^-1. At the same time, the retention mechanism between protein and G-F-ZrO2 stationary phase was also discussed.