High-Speed Two Dimensional Separations Using Micro Free Flow Electrophoresis

在化学系化学测量和成像计划的支持下，明尼苏达大学双子城分校的Michael Bowser教授和他的团队将使用微型自由流动电泳(UFFE)作为2D分离的第二阶段。FFE是一种连续分析技术，能够在分析物峰从第一个分离柱上洗脱时进一步分离它们。连续分析消除了由于采样不足而在以前的2D分离中遇到的限制。与毛细管电泳色谱(CE)或纳米液相色谱(NLC)的联用被认为是直接的，不需要复杂的阀门或时间。预计与UFFE的结合将使这些技术的峰容量增加20到30倍，而不会牺牲第一维分离的效率或分离时间。在15分钟的分离时间内，总2D峰容量有望达到2,000。计划进行实验，以充分表征使用该方法可能的多种分离模式组合的峰容量和兼容性。为了展示高速2D分离的潜在能力，CE-UFFE将被纳入在线微透析分析中，用于测量体内的神经化学动力学。在线微透析需要对复杂混合物进行高速分离，这使得该分析成为展示本提案中开发的2D分离优势的理想应用。该方法的技术优势在于在不牺牲分离时间的情况下增加了峰容量。因此，二维UFFE具有被广泛采用的潜力，可以提高许多研究领域的研究能力和效率。除了实际优势之外，对2D-UFFE的探索将为分离科学的基本原理提供独特的见解。密歇根大学双子城的研究人员还打算设计本提案中描述的研究项目，目的是通过智力丰富、指导和培训机会、纳入代表性不足的群体和加强基础设施，最大限度地提高其对社会的影响。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Michael Bowser at University of Minnesota-Twin Cities and his group will use micro free flow electrophoresis (uFFE) as the second stage in 2D separations. FFE is a continuous analysis technique capable of further separating analyte peaks as they elute off the first separation column. Continuous analysis removes limitations encountered in previous 2D separations caused by under sampling. Coupling with capillary electrophoresis (CE) or nano liquid chromatography (nLC) is expected to be straightforward with no complicated valving or timing required. It is anticipated that coupling with uFFE will increase the peak capacity of these techniques 20 to 30-fold with no sacrifice of efficiency or separation time in the first dimension separation. Total 2D peak capacities are expected to reach 2,000 in separation times of 15 min. Experiments are planned to fully characterize the peak capacity and compatibility of the many combinations of separation modes possible using this approach. As a demonstration of the potential power of high speed 2D separations CE-uFFE will be incorporated into online microdialysis assays for measuring in vivo neurochemical dynamics. Online microdialysis requires high-speed separations of complex mixtures, making this assay an ideal application to demonstrate the advantages of the 2D separations developed in this proposal. The technical benefit of the proposed approach lies in its increased peak capacity without sacrificing separation time. 2D uFFE therefore has the potential to be adopted widely, improving research capabilities and efficiency in many research areas. Beyond practical advantages, exploration into 2D-uFFE will provide unique insight into the fundamentals of separation science. The researchers at UM-twin cities also intend to design the research project described in this proposal with the goal of maximizing its impact on society through intellectual enrichment, mentoring and training opportunities, inclusion of underrepresented groups and infrastructure enhancement.