葛根提取物高效液相色谱分析方法的优化

针对葛根提取物的复杂体系,分别从色谱模式、流动相、添加剂以及梯度洗脱条件几个方面对其高效液相色谱分析 方法进行了优化。其中梯度洗脱条件的优化是根据各组分在5次线性梯度下的保留时间并借助于CSASS软件快速准确地计 算出各组分的液相色谱保留参数a,c值以及峰形参数σ和τ,在此基础上使用CSASS软件对葛根提取物的色谱分离情况进行 高精度仿真预测,从而优化了梯度洗脱条件。考察了优化条件下的分析方法的精密度和重现性,结果表明该方法稳定、可 靠、重现性好。

Optimization of High Performance Liquid Chromatographic Method for Analysis of Kudzu Root Crude Extract

Kudzu root is an important traditional Chinese herb. Its crude extract has been used in the treatment of hypertension and angina pectoris in China. Several parameters for high performance liquid chromatography (HPLC) analytical method for kudzu root extract, including HPLC model, mobile phase, additive and gradient conditions have been optimized. The HPLC retention parameters a and c, and the peak shape parameters sigma and tau of 25 chromatographic peaks were obtained accurately and rapidly using five linear gradients and were calculated using CSASS software. The CSASS software was then used to simulate the gradient conditions in the experiments, and the optimized condition was obtained. The comparison of the simulated and real chromatogram profiles showed that the potential of simulation using the CSSASS software as compared to that of the real experimental conditions was precise. Reversed-phase HPLC model and the mobile phase of (A) acetonitrile containing 0.1% (v/v) acetic acid and (B) water containing 0.1% (v/v) acetic acid were used. The optimized gradient profile showed a linear increase as follows: from 5% A to 35% A at 0 - 33 min; from 35% A to 100% A at 33 - 50 min, and then the HPLC system was held for 5 min. The reproducibility and precision of the method were investigated. The relative standard deviations (RSDs) of retention time, peak height, and peak area were less than 0.25%, 11%, and 4.9%, respectively, which showed that the method was stable, reliable, and reproducible.