Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC, Delft, The Netherlands.
J Chromatogr A. 2012 May 25;1239:22-34. doi: 10.1016/j.chroma.2012.03.042. Epub 2012 Mar 20.
Adsorption equilibria of the saccharides d-glucose, d-galactose, l-arabinose, lactose and a sugar acid were measured on gel-type sulfonated poly (styrene-co-divinylbenzene) strong cation exchange resins in a high-throughput (HT) 96-well plate batch uptake mode using a pipetting robot at 25 °C. Four different ionic forms, Ca(2+), K(+), Na(+), and H(+) were used. Single component adsorption isotherms were determined in a concentration range of 10-240 mg ml(-1). Multicomponent experiments were performed to investigate competitive adsorption in a concentration range of 10-120 mg ml(-1). A qualitative investigation on competitive and cooperative effects was performed. All sugar isotherms showed a linear behavior except for the sugar acid which showed an unfavorable (anti-Langmuir) behavior in the high concentration ranges. Selectivity values were determined from the binary mixture partition coefficient (K) values of each component. This HT 96-well plate batch uptake method proves to be less laborious and consumes less time and material compared to the frontal analysis and adsorption-desorption methods where column experimentation is used. Ternary mixture separation of arabinose and the sugar acid from glucose showed K(+) and Ca(2+) loaded resins having the best selectivity (DIAION Ca(2+) 2.01 and 1.78 for l-arabinose/d-glucose and sugar acid/l-arabinose respectively), similarly Purolite K(+) loaded resin for the lactose separation from glucose and galactose (1.17 for lactose/d-glucose). Column experiments were performed to validate the batch uptake experiments. The static binding results could easily be translated to the column experiments with good agreement. Finally, adding to the validity of the approach, binary and ternary fixed-bed experiments were well described by a dynamic mathematical chromatographic model using the parameters obtained from the binary-component isotherm data.
在 25°C 下,使用移液机器人在高通量 (HT) 96 孔板分批吸附模式下,在凝胶型磺化聚苯乙烯-二乙烯基苯强阳离子交换树脂上测量了葡萄糖、半乳糖、阿拉伯糖、乳糖和糖酸等单糖的吸附平衡。使用了四种不同的离子形式:Ca(2+)、K(+)、Na(+)和 H(+)。在 10-240mg/ml 的浓度范围内确定了单一组分吸附等温线。在 10-120mg/ml 的浓度范围内进行了多组分实验以研究竞争吸附。进行了定性的竞争和协同作用研究。除了糖酸在高浓度范围内表现出不利的(反朗缪尔)行为外,所有糖的等温线都表现出线性行为。从每个组分的二元混合物分配系数 (K) 值确定了选择性值。与使用柱实验的前沿分析和吸附-解吸方法相比,这种 HT 96 孔板分批吸附方法证明劳动强度更低,耗时和材料更少。从葡萄糖中分离阿拉伯糖和糖酸的三元混合物显示,负载 K(+)和 Ca(2+)的树脂具有最佳选择性(DIAION Ca(2+) 对 l-阿拉伯糖/葡萄糖和糖酸/l-阿拉伯糖的分别为 2.01 和 1.78),同样 Purolite K(+)负载树脂对乳糖与葡萄糖和半乳糖的分离也具有最佳选择性(乳糖/葡萄糖为 1.17)。进行了柱实验以验证分批吸附实验。静态结合结果可以很容易地转化为柱实验,并且具有良好的一致性。最后,通过使用从二元组分等温线数据获得的参数,使用动态数学色谱模型很好地描述了二元和三元固定床实验,这增加了该方法的有效性。