反相高效液相色谱三元流动相台阶梯度分离条件的快速优化方法  

Fast Optimization of Stepwise Gradient Conditions for Ternary Mobile Phase in Reversed-Phase High Performance Liquid Chromatography

作　　者：单亦初[1] 张玉奎[1] 赵瑞环[1] 

机构地区：[1]中国科学院大连化学物理研究所国家色谱研究分析中心,辽宁大连116011

出　　处：《色谱》2002年第4期289-294,共6页Chinese Journal of Chromatography

基　　金：国家自然科学基金资助项目 (2 0 175 0 2 7)

摘　　要：根据溶质在色谱柱内的迁移规律 ,建立了一种利用线性梯度实验快速获得溶质保留值方程系数 ,然后以串行色谱响应函数为优化指标进行多台阶梯度分离条件优化的方法。与利用等度实验获得保留值方程的方法相比 ,该法可以大大缩短优化时间。通过该方法对芳香胺和衍生化氨基酸样品进行了分离 ,获得了满意的分离度 ,表明该方法的预测精度很好。In high performance liquid chromatography, it is necessary to apply multi composition gradient elution for the separation of complex samples such as environmental and biological samples. Multivariate stepwise gradient elution is one of the most efficient elution modes, because it combines the high selectivity of multi composition mobile phase and shorter analysis time of gradient elution. In practical separations, the separation selectivity of samples can be effectively adjusted by using ternary mobile phase. For the optimization of these parameters, the retention equation of samples must be obtained at first. Traditionally, several isocratic experiments are used to get the retention equation of solute. However, it is time consuming especially for the separation of complex samples with a wide range of polarity. A new method for the fast optimization of ternary stepwise gradient elution was proposed based on the migration rule of solute in column. First, the coefficients of retention equation of solute are obtained by running several linear gradient experiments, then the optimal separation conditions are searched according to the hierarchical chromatography response function which acts as the optimization criterion. For each kind of organic modifier, two initial linear gradient experiments are used to obtain the primary coefficients of retention equation of each solute. For ternary mobile phase, only four linear gradient runs are needed to get the coefficients of retention equation. Then the retention times of solutes under arbitrary mobile phase composition can be predicted. The initial optimal mobile phase composition is obtained by resolution mapping for all of the solutes. A hierarchical chromatography response function is used to evaluate the separation efficiencies and search the optimal elution conditions. In subsequent optimization, the migrating distance of solute in the column is considered to decide the mobile phase composition and sustaining time of the latter steps unti

关 键 词：三元流动相 分离条件 优化方法 反相高效液相色谱法 线性梯度 台阶梯度 串行色谱响应函数 

分 类 号：O657.72[理学—分析化学] O658[理学—化学]