Guijt R M, Baltussen E, van der Steen G, Frank H, Billiet H, Schalkhammer T, Laugere F, Vellekoop M, Berthold A, Sarro L, van Dedem G W
Laboratory of Analytical Biotechnology, Kluyver Institute for Biotechnology, Delft University of Technology, The Netherlands.
Electrophoresis. 2001 Aug;22(12):2537-41. doi: 10.1002/1522-2683(200107)22:12<2537::AID-ELPS2537>3.0.CO;2-C.
Microchip capillary electrophoresis (CE) with integrated four-electrode capacitively coupled conductivity detection is presented. Conductivity detection is a universal detection technique that is relatively independent on the detection pathlength and, especially important for chip-based analysis, is compatible with miniaturization and on-chip integration. The glass microchip structure consists of a 6 cm etched channel (20 microm x 70 microm cross section) with silicon nitride covered walls. In the channel, a 30 nm thick silicon carbide layer covers the electrodes to enable capacitive coupling with the liquid inside the channel as well as to prevent interference of the applied separation field. The detector response was found to be linear over the concentration range from 20 microM up to 2 mM. Detection limits were at the low microM level. Separation of two short peptides with a pI of respectively 5.38 and 4.87 at the 1 mM level demonstrates the applicability for biochemical analysis. At a relatively low separation field strength (50 V/cm) plate numbers in the order of 3500 were achieved. Results obtained with the microdevice compared well with those obtained in a bench scale CE instrument using UV detection under similar conditions.
介绍了一种集成四电极电容耦合电导检测的微芯片毛细管电泳(CE)技术。电导检测是一种通用的检测技术,相对独立于检测路径长度,对于基于芯片的分析尤为重要,它与小型化和芯片集成兼容。玻璃微芯片结构由一个6厘米长的蚀刻通道(横截面为20微米×70微米)组成,通道壁覆盖有氮化硅。在通道中,一层30纳米厚的碳化硅层覆盖电极,以实现与通道内液体的电容耦合,并防止施加的分离场产生干扰。发现检测器响应在20微摩尔至2毫摩尔的浓度范围内呈线性。检测限处于低微摩尔水平。在1毫摩尔浓度下对两种pI分别为5.38和4.87的短肽进行分离,证明了该技术在生化分析中的适用性。在相对较低的分离场强(50伏/厘米)下,实现了约3500的塔板数。在类似条件下,使用该微器件获得的结果与在台式CE仪器中使用紫外检测获得的结果相当。