反相高效液相色谱法分离蛋白质的研究

Study of Protein Separation by Reversed Phase High Performance Liquid Chromatography

采用反相高效液相色谱法考察了几种大孔硅胶烷基键合固定相在等度淋洗条件下进行蛋白质分离的色谱性能。研究了冲洗剂中有机溶剂异丙醇的浓度、离子对酸（ＴＦＡ）浓度对蛋白质保留时间的影响。探讨了蛋白质在ＲＰ－ＨＰＬＣ中的保留机理。结果表明，大孔硅胶（２０～３０ｎｍ）短链（Ｃ４和Ｃ８）烷基键合固定相适合蛋白质的分离。

Reversed phase high performance liquid chromatography (RP HPLC) of protein on large pore silica(20 30nm) n alkyl bonded(C 4 and C 8) phase columns was studied using a trifluoroacetic acid (TFA)/2 propanol mobile phase system. The influence of concentration of organic modifier (2 propanol) and ion pairing acid(TFA) on the protein retention time was examined. The results showed that the plot of the retention time vs. 2 propanol concentration in mobile phase was U shaped. The mechanism of protein in reversed phase high performance liquid chromatography is also discussed.