摘要
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
The reversed phase chromatographic method consisting of acetic acid water as a new mobile phase for separating biopolymers on ODS column is presented The new mobile phase may overcome the weak points of irreversible adsorption in chromatographic separation to some degree and difficulty in concentrating proteins with lyophylization and reduce the three component (organic solvent, ion pairing agent and water) of typical mobile phase to two component system. Based on the retention and Z (the number of solvent molecules involving in the displacement from stationary phase surface ), log I (a constant relating to the affinity of 1 mol of solute to the bonded stationary phase ), and j (a constant relating to the affinity of 1 mol of solvent to the bonded stationary phase ), the elution strength of acetic acid water mobile phase was found to be stronger than that of methanol water TFA From a good linear relationship between log I and Z , it can be concluded that the interaction force between proteins and RPLC stationary phase with acetic acid water as mobile phase is non selective hydrophobic interaction forces In addition, the molecular conformations of protein in acetic acid water and methanol trifluoroacetic acid water were found to be different.
出处
《色谱》
CAS
CSCD
北大核心
2000年第6期487-490,共4页
Chinese Journal of Chromatography
基金
国家自然科学基金资助课题! (2 96 75 0 17
39880 0 0 3)
关键词
反相高效液相色谱法
生物大分子
保留行为
分离
reversed phase high performance liquid chromatography
stoichiometric displacement
biopolym$
