提出了以醋酸 水作为流动相的体系中 ,在ODS柱上分离生物大分子的反相高效液相色谱 (RPLC)方法。实验结果表明 ,醋酸 水的洗脱能力强于甲醇 水 三氟醋酸体系 ,在一定程度上克服了色谱分离中一些蛋白质的不可逆吸附且具有便于冷冻干燥的优点。用参数Z(1mol溶剂化溶质被溶剂化固定相吸附时从两者接触表面释放出置换剂的摩尔总数 ) ,logI(与 1mol溶质对固定相亲和势有关的常数 )和 j(与 1mol溶剂对固定相亲和势有关的常数 )对 9种蛋白质在此流动相体系中的保留进行了表征。该流动相使在RPLC中用于分离生物大分子的三元流动相体系成为二元流动相体系 ,更适合于理论研究。同时得出结论 :在醋酸 水体系中 ,蛋白质与固定相之间的作用力主要为非选择性作用力 ;蛋白质在醋酸 水和甲醇 水 三氟醋酸两种流动相体系中的分子构象变化不同 A reversed-phase high performance liquid chromatography (RPLC) method was developed for the separation of biological macromolecules on ODS column using acetic acid water as the mobile phase. The experimental results show that the elution ability of acetic acid water is stronger than that of methanol-water trifluoroacetic acid system, which overcomes the irreversible adsorption of some proteins in chromatographic separation to a certain extent and has the advantages of easy freeze-drying. The log I (constant related to 1 mol of solute to the stationary phase affinity) and j (relative to 1 mol of solvent) were determined using parameter Z (the total number of moles of displacer released from the contact surfaces of the solute of 1 mol with the solvated stationary phase) Fixed phase affinity-related constants) The retention of nine proteins in this mobile phase system was characterized. The mobile phase makes the ternary mobile phase system used to separate biological macromolecules in RPLC into a binary mobile phase system, which is more suitable for theoretical research. At the same time, it is concluded that the interaction between protein and stationary phase is mainly non-selective in aqueous acetic acid system. The molecular conformational changes of the two mobile phases of acetic acid water and methanol water trifluoroacetic acid are different