Development of comprehensive two-dimensional HPLC for high-performance separations in the future

开发用于未来高性能分离的综合二维 HPLC

基本信息

批准号：
14340234
负责人：
TANAKA Nobuo
金额：
$ 7.55万
依托单位：
Kyoto Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

The objective of this research project was to increase peak capacity of HPLC by developing simple and comprehensive two dimensional (2D) HPLC. By employing a monolithic silica-C18 column at the second dimension (2nd-D), we were able to obtain the following results.(1) We proved the concept of simple and comprehensive 2D-HPLC in a reversed-phase-reversed-phase mode by using a particle-packed column of fluoroalkyl-bonded silica as a first-dimension (1st-D) column, and C18- or pentabromobenzyloxypropyl-bonded monolithic silica as a 2nd-D column.(2) By utilizing the difference in selectivity provided by organic solvents, 2D-HPLC was carried out using monolithic silica C18 columns both at 1st-D and at 2nd-D.(3) 2D-HPLC was carried out by using an ion-exchange column packed with polymer particles at 1st-D and a monolithic silica-C18 column or a monolithic silica capillary column at 2nd-D for peptide separations.The use of these systems produced (1)peak capacity of up to 1100 in methanol-water mobile phase, (2)peak capacity of 700 or greater for hydrocarbons and benzene derivatives, respectively. The ion-exchange-reversed-phase 2D-HPLC system provided (3)peak capacity of 700 or greater for BSA digest with a short monolithic silica column at 2nd-D, and peak capacity of about 500 for BSA digest by using a 0.2 mmlD, 10 cm long capillary column at 2nd-D, respectively. Optimization of 2D-HPLC conditions were attempted to show the possibility of achieving peak capacity of greater than 2000 in one hour. The flow rate, retention factors of solutes, and column length at 2nd-D were shown to dominate the peak capacity of a 2D-HPLC system. Monolithic silica columns were found to be best suited for 2D-HPLC based on their high permeability and high performance at high linear velocity of mobile phase.

该研究项目的目的是通过开发简单而全面的二维（2D）HPLC来提高HPLC的峰值能力。通过在第二维（2nd-d）上使用整体硅C18柱，我们能够获得以下结果。（1）我们通过使用相反的相位相位阶段模式证明了简单且全面的2D-HPLC概念，该概念是通过使用粒子包装的Flooroaloaloaloaloaloaloalokyl-Bond-Bond silica（1St-Dimeension（1St-D）列（1St-D）和C1ST-D）的概念，或者五氢苯甲酰氧丙基键键单层二氧化硅作为第二列列。（2）通过利用有机溶剂提供的选择性差异，使用单层二氧化硅C18列在1st-d和2nd-d-hplc colled cartect and contere contect and c18列中进行了2D-HPLC。（3）2D。 1st-D和一个单片二氧化硅C18柱或在2nd-D处的单片二氧化硅毛细管柱用于肽分离。（1）在甲醇 - 含水流动相位（1）高达1100个峰值的峰值容量，（2）分别用于700或更大的峰值峰值和更大的氢碳和苯衍生物。离子交换 - 反向相2D-HPLC系统（3）BSA摘要的峰值容量为700或更高的峰值，分别使用0.2 MMLD，分别在2ND-D-DND上使用0.2 MMLD，使用0.2 MMLD，使用0.2 MMLD，使用0.2 MMLD，使用0.2 MMLD，使用0.2 MMLD。试图优化2D-HPLC条件，以表明在一小时内达到2000年峰值容量的可能性。显示流速，溶质的保留因子和2nd-D时的柱长度占据了2D-HPLC系统的峰值容量。发现单片二氧化硅柱最适合2D-HPLC，基于它们在流动相高的线性速度下的高渗透性和高性能。