Department of Chemistry, Center for Biotechnology and Drug Design, Georgia State University, 38 Peachtree Center Ave, Atlanta, GA 30303, USA.
J Chromatogr A. 2010 Jan 22;1217(4):530-9. doi: 10.1016/j.chroma.2009.11.082. Epub 2009 Dec 3.
A surfactant-bound monolithic stationary phase based on the co-polymerization of 11-acrylamino-undecanoic acid (AAUA) is designed for capillary high performance liquid chromatography (HPLC). Using D-optimal design, the effect of the polymerization mixture (concentrations of monomer, crosslinker and porogens) on the chromatographic performance (resolution and analysis time) of the AAUA-EDMA monolithic column was evaluated. The polymerization mixture was optimized using three proteins as model test solutes. The D-optimal design indicates a strong dependence of chromatographic parameters on the concentration of porogens (1,4-butanediol and water) in the polymerization mixture. Optimized solutions for fast separation and high resolution separation, respectively, were obtained using the proposed multivariate optimization. Differences less than 6.8% between the predicted and the experimental values in terms of resolution and retention time indeed confirmed that the proposed approach is practical. Using the optimized column, fast separation of proteins could be obtained in 2.5 min, and a tryptic digest of myoglobin was successfully separated on the high resolution column. The physical properties (i.e., morphology, porosity and permeability) of the optimized monolithic column were thoroughly investigated. It appears that this surfactant-bound monolith may have a great potential as a new generation of capillary HPLC stationary phase.
基于 11-丙烯酰胺基-1-十一烷酸（AAUA）共聚体制备的表面活性剂键合整体固定相，用于毛细管高效液相色谱（HPLC）。采用 D-最优设计，考察了聚合混合物（单体、交联剂和致孔剂浓度）对 AAUA-EDMA 整体柱色谱性能（分辨率和分析时间）的影响。使用三种蛋白质作为模型测试溶质对聚合混合物进行了优化。D-最优设计表明色谱参数强烈依赖于聚合混合物中致孔剂（1,4-丁二醇和水）的浓度。采用提出的多元优化方法，分别得到了快速分离和高分辨率分离的优化溶液。在分辨率和保留时间方面，预测值与实验值之间的差异小于 6.8%，这确实证实了所提出的方法是实用的。使用优化的柱子，可以在 2.5 分钟内快速分离蛋白质，并且可以在高分辨率柱子上成功分离肌红蛋白的酶解产物。对优化后的整体柱的物理性质（即形态、孔隙率和渗透性）进行了彻底研究。这种表面活性剂键合整体柱似乎具有作为新一代毛细管 HPLC 固定相的巨大潜力。
