利用HCI程序确定儿茶酚化合物的反相高效液相色谱最佳分离条件  

Optimum Separation Conditions of Catechin Compounds by HCI Program in Reversed-Phase High Performance Liquid Chromatography

作　　者：金银哲[1] 卢敬昊[1] 

机构地区：[1]仁荷大学化工系及生物分离技术研究中心

出　　处：《色谱》2006年第5期466-470,共5页Chinese Journal of Chromatography

摘　　要：有效地确定了反相高效液相色谱分离儿茶酚化合物的最佳条件。在水和甲醇的二元流动相里分别加入乙酸缓冲液,利用基于lnk=lnkw+SF,k=A+B/F,lnk=L+MF+NF2(F是流动相中有机物甲醇的体积分数)等保留因子的一次或二次方程式的塔板理论得到色谱分离结果;利用保留原理得到等度和梯度洗脱的最佳条件。得出最佳初始流动相是含0.1%乙酸的水和含0.1%乙酸的甲醇(体积比为75∶25)的混合溶液;梯度洗脱条件:初始流动相保持15m in,然后用10m in的时间将上述二元流动相的体积比线性变换成50∶50,直到完成全部分离。通过实验证实该计算结果与实验值相近。An efficient optimization method was used to separate catechin compounds by reversed-phase high performance liquid chromatography （RP-HPLC）. The binary mobile phase of water and methanol was utilized with the buffer of acetic acid （AA）. The elution profiles were calculated by the plate theory based on the linear and quadratic equations of retention factor, Ink = In kw ＋ SF, k = A ＋ B/F, Ink = L ＋ MF ＋ NF^2, where F is the volume fraction of methanol in the mobile phase. The retention theory was modified to calculate the elution profile in both isocratic and gradient modes. Based on the retention theory, elution profiles were predicted by introducing the concept of solute migration in the mobile phase with the linear and quadratic dependence of Ink in terms of the organic modifier content. Using the HCI program （ a software designed by Inha University）, the recommended experimental conditions of mobile phase composition and gradient step were suggested, and the elution profiles calculated by the quadratic relationship of Ink showed better coincidence with the experimental data than the linear correlation did. The calculated results of mobile phase condition for separation of catechin compounds suggested that the mobile phase composition was 0. 1% AA in water/0. 1% AA in methanol, 75/25 （v/v）, then after 15 min, the composition was linearly changed to 50/50 （v/v） in 10 min and held at the isocratic mode to the end. In the experimental conditions, the agree- ment between the experimental elution profiles and the calculated values of eluted concentration was relatively good.

关 键 词：反相高效液相色谱 儿茶酚化合物 HCI程序 保留原理 流动相组成 

分 类 号：O658[理学—分析化学]