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。 The chromatographic properties of several macroporous silica gel alkyl-bonded stationary phases for protein separation under isocratic elution conditions were investigated by RP-HPLC. The concentration of isopropanol, the organic solvent in the irrigant, and the effect of ion-pair acid (TFA) on the protein retention time were studied. The retention mechanism of protein in RP-HPLC was discussed. The results showed that macroporous silica gel (20-30 nm) short chain (C4 and C8) alkyl-bonded phases were suitable for protein separation.