Department of Chemistry and Biochemistry, Florida State University, Tallahassee, 32306, USA.
J Chromatogr A. 2010 Jul 9;1217(28):4743-8. doi: 10.1016/j.chroma.2010.05.023.
A microfluidic device is presented that performs electrophoretic separation coupled with fraction collection. Effluent from the 3.5 cm separation channel was focused via two sheath flow channels into one of seven collection channels. By holding the collection channels at ground potential and varying the voltage ratio at the two sheath flow channels, the separation effluent was directed to either specific collection channels, or could be swept past all channels in a defined time period. As the sum of the voltages applied to the two sheath flow channels was constant, the electric field remained at 275 V/cm during the separation regardless of the collection channel used. The constant potential in the separation channel allowed uninterrupted separation for late-migrating peaks while early-migrating peaks were being collected. To minimize the potential for carryover between fractions, the device geometry was optimized using a three-level factorial model. The optimum conditions were a 22.5° angle between the sheath flow channels and the separation channel, and a 350 μm length of channel between the separation outlet and the fraction channels. Using these optimized dimensions, the device performance was evaluated by separation and fraction collection of a fluorescently-labeled amino acid mixture. The ability to fraction collect on a microfluidic platform will be especially useful during automated or continuous operation of these devices or to collect precious samples.
本文介绍了一种微流控装置,可实现电泳分离与馏分收集的偶联。3.5cm 分离通道的流出物通过两个鞘流通道聚焦到七个收集通道之一。通过将收集通道保持在接地电位,并改变两个鞘流通道的电压比,可以将分离流出物引导到特定的收集通道,或者可以在定义的时间段内扫过所有通道。由于施加到两个鞘流通道的电压总和保持不变,因此无论使用哪个收集通道,分离过程中的电场都保持在 275V/cm。分离通道中的恒电位允许在收集较早迁移的峰的同时,不间断地进行较晚迁移的峰的分离。为了最大限度地减少馏分之间的交叉污染的可能性,使用三水平析因模型优化了器件几何形状。最佳条件是鞘流通道与分离通道之间的夹角为 22.5°,以及分离出口与馏分通道之间的通道长度为 350μm。使用这些优化的尺寸,通过荧光标记的氨基酸混合物的分离和馏分收集评估了器件性能。在这些设备的自动化或连续操作期间,或者在收集珍贵样品时,能够在微流控平台上进行馏分收集将特别有用。