Departamento de Química Analítica, Universidad de Valencia, c/Dr. Moliner 50, 46100 Burjassot, Spain.
Anal Bioanal Chem. 2012 Dec;404(10):2973-84. doi: 10.1007/s00216-012-6387-7. Epub 2012 Sep 25.
Two approaches are proposed to measure the effect of different experimental factors (such as the modifier concentration and temperature) on the elution strength and peak shape in reversed-phase liquid chromatography, which quantify the percentage change in the retention factor and peak width (referred to the weakest conditions) per unit change in the experimental factor. The approaches were applied to the separation of a set of flavonoids with aqueous micellar mobile phases of the surfactant Brij-35 (polyoxyethylene(23)dodecanol), in comparison with acetonitrile-water mixtures, using an Eclipse XDB-C18 column. The particular interaction of each flavonoid with the oxyethylene chains of Brij-35 molecules (adsorbed on the stationary phase or forming micelles) changed the elution window, distribution of chromatographic peaks and partitioning kinetics, depending on the hydroxyl substitution in the aromatic rings of flavonoids. At 25 °C, peak shape with Brij-35 mobile phases was significantly poorer with regard to acetonitrile-water mixtures. At increasing temperature, the efficiency of Brij-35 increased, approaching at 80 °C the values obtained at equilibrium conditions, already reached with acetonitrile at 25 °C.
提出了两种方法来衡量不同实验因素（如修饰剂浓度和温度）对反相液相色谱中洗脱强度和峰形的影响，这两种方法量化了保留因子和峰宽（相对于最弱条件）每单位实验因素变化的百分比变化。该方法应用于一组黄酮类化合物的分离，采用表面活性剂 Brij-35（聚氧乙烯（23）十二醇）的水胶束流动相，与乙腈-水混合物进行比较，使用 Eclipse XDB-C18 柱。每种黄酮类化合物与 Brij-35 分子的氧乙烯链之间的特殊相互作用（吸附在固定相上或形成胶束）改变了洗脱窗口、色谱峰分布和分配动力学，这取决于黄酮类化合物芳环上的羟基取代。在 25°C 时，与乙腈-水混合物相比，Brij-35 流动相的峰形明显较差。随着温度的升高，Brij-35 的效率增加，在 80°C 时接近在平衡条件下获得的值，而在 25°C 时已达到乙腈的水平。
