高效液相色谱用线性梯度洗脱求保留值方程

The Prediction of Retention Equation in the Reversed-Fhase High Performance Liquid Chromatography Using the Linear Gradient Eiution

提出了采用线性梯度洗脱求取未知样品中溶质保留值方程的原理及其计算方法,用光二极管阵列扫描检测器解决了梯度洗脱条件下色谱峰的归宿。实验证明:用所得的保留值方程预测等度洗脱下容量因子k′的相对误差不大于3％,而用梯度洗脱下的半峰宽规律可以估算未知样品容量因子k′的分布范围,以解决对未知样品是采用等度或梯度洗脱方式。最后,借助计算机仿真方法用于样品的优化,获得了满意的结果。

The principles and algorithm of prediction of retention equation in reversed-phase HPLC for a mixture with unknown constituents using linear gradient elution has been presented.The fast scanning photodiode array detector (PAD) has been employed to monitor the position of solutes,in the various gradient runs.It has been shown experimentally that the relative errors of isocratic data predicted by the retention equation obtained from iterative algorithm were less than 3%.In addition,the isocratic data predicted by the retention equation obtained from the correlation of half-height width and retention time can be used to estimate the range of capacity factors in the mixture to select elution mode for the samples.The retention equation obtained from the linear gradient elution has been used for the optimization of the mixture separation.The results were in good agreement with those of the actual runs.