Development of ultra high-resolution capillary liquid phase chromatographic system

1. 采用季铵离子动态修饰硅胶整体毛细管柱测定海水样品中的溴化物。将十六烷基三甲基铵等季铵离子动态引入单片二氧化硅表面。第一层改性剂通过静电相互作用引入，第二层改性剂通过疏水相互作用引入。后一层作为阴离子交换的场所。改进的整体硅胶毛细管柱可用于无机阴离子的快速分离。五种阴离子的真实混合物在几分钟内就分离出来了。在洗脱液中加入少量的改性剂，提高了保留时间的重复性。海水样品可直接注入制备的毛细管柱，溴化物的测定值为63 mg/L.2。采用微柱液相色谱法（LC）研究了硼酸预柱在水溶液中痕量测定双酚a（4,4′-异丙基，多酚或双酚a）中的应用。BPA通过填充TSKgel硼酸盐的前柱富集，然后注入（反冲）到ODS分离柱中。与传统注射器注射相比，硼酸钠柱前注射获得了更好的峰形，计算的理论板数提高了约2倍。保留时间、峰面积和峰高的相对标准偏差分别为0.12、3.0和1.8%。该系统在工作样品浓度和样品体积范围内呈线性（R2 > 0.99），当富集0.2 ml样品时，检测限为0.09μgl^<-1> (S/N=3)。该方法的回收率在107 ~ 123%之间，适用于市售瓶装饮料样品中BPA的测定。就双酚a在水样中的富集而言，硼酸盐预柱被证明比ODS或C30预柱具有更好的灵敏度。建立了一种新型的酶固定玻璃微珠微柱，用于在线、简单、快速的蛋白质消化，并通过反相高效液相色谱分离多肽。采用氨丙基控制孔玻璃，首先与戊二醛反应，然后在磷酸缓冲液存在下与胰蛋白酶反应，制备了这种廉价且易于制备的酶固定化微柱。牛血清白蛋白（BSA）的胰酶消化是通过实验室制作的胰酶固定柱进行的，然后通过反相高效液相色谱分离胰酶片段。发现多肽的分离与使用在溶液中进行常规酶解的样品所做的相同。少

1. Monolithic silica capillary columns dynamically modified with quaternary ammonium ions were evaluated for the determination of bromide in seawater samples. A quaternary ammonium ion such as cetyltrimethylammonium was dynamically introduced onto monolithic silica surfaces. The first layer of the modifier was introduced by electrostatic interaction, whereas the second layer was introduced by hydrophobic interaction. The latter layer worked as the anion-exchange sites. The modified monolithic silica capillary columns could be used for rapid separation of inorganic anions. Separation of authentic mixture of five anions was achieved within a few minutes. The addition of small amount of the modifier in the eluent improved the repeatability of the retention time. Seawater samples could be directly injected onto the prepared capillary columns, and bromide could be determined to be 63 mg/L.2. The application of a boronate precolumn in the trace determination of bisphenol A(4,4'-isopropyliden … More ediphenol or BPA) in aqueous solution was investigated by microcolumn liquid chromatography (LC). BPA was enriched by a precolumn packed with TSKgel boronate and it was then injected (back-flushed) into an ODS separation column. Comparing to a conventional syringe injection, a better peak shape was obtained via the boronate precolumn injection, and the calculated theoretical plate number improvement was approximately 2 folds. The relative standard deviations for the retention time, peak area and peak height were 0.12, 3.0 and 1.8%, respectively. The system is linear (R2 > 0.99) within the working sample concentration and sample-volume ranges, and it presents a detection limit of 0.09μgl^<-1> (S/N=3) when a 0.2 ml sample was enriched. Recovery values of the proposed method were between 107 and 123%, and this method was applied satisfactorily to the determination of BPA in commercially available bottled drink samples. The boronate precolumn was proven to produce better sensitivity than that of an ODS or C30 precolumn as long as the enrichment of BPA in aqueous samples is concerned.3. A novel enzyme-immobilized glass beads based micro column was developed for the on-line, simple and swift protein digestion, followed by peptide separation via reversed-phase HPLC. This inexpensive and easy-made enzyme-immobilized micro column was prepared by using aminopropyl-controlled pore glass that was first reacted with glutaraldehyde followed by reaction with trypsin under the presence of phosphate buffer. Tryptic digestion of bovine serum albumin (BSA) was carried out by simply passing the pre-treated protein solution through the laboratory made trypsin-immobilized column, then followed by separation of tryptic fragments via reversed-phase HPLC. The separation of peptides was found to be equivalent to those done by using a sample which was underwent a conventional enzymatic protein digestion performed in solution. Less