Ishida Akihiko, Natsume Masamichi, Kamidate Tamio
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, Nishi 8, Kita 13, Sapporo 060-8628, Japan.
J Chromatogr A. 2008 Dec 12;1213(2):209-17. doi: 10.1016/j.chroma.2008.10.037. Epub 2008 Oct 17.
A microchip pressure-driven liquid chromatography (LC) with a packed column and an electrochemical flow cell has been developed by using polystyrene (PS) and poly(dimethylsiloxane) (PDMS). The cylindrical separation column with packed octadecyl silica particles was fabricated in the PS substrate. The three electrode system (working, reference, and counter electrode) for amperometric detection was fabricated onto the PS substrate, using the Au deposition, photolithography, and chemical etching. The detector flow cell was formed by sealing the electrode system with a PDMS chip containing a channel. In this flow cell, the effect of working electrode width (in the direction of flow) on chromatographic parameters, such as peak width and peak resolution were studied in electrode width ranging 50-5,000 microm. The effect of electrode width on sensitivity (current intensity, current density, and S/N ratio) was also examined. The sensitivity was discussed by simulating the concentration profile generated around the working electrode. The effects of the column packing size and the column size on the separation efficiency were examined. In this study, a good separation of three catechins was successfully achieved and the detection limits for (+)-catechin, epicatechin, and epigallocatechin gallate were 350, 450, and 160 nM, respectively.
利用聚苯乙烯(PS)和聚二甲基硅氧烷(PDMS)开发了一种带有填充柱和电化学流通池的微芯片压力驱动液相色谱(LC)。在PS基板上制作了填充十八烷基硅胶颗粒的圆柱形分离柱。采用金沉积、光刻和化学蚀刻技术,在PS基板上制作了用于安培检测的三电极系统(工作电极、参比电极和对电极)。通过用含有通道的PDMS芯片密封电极系统形成检测器流通池。在这个流通池中,研究了工作电极宽度(沿流动方向)对色谱参数(如峰宽和峰分辨率)的影响,电极宽度范围为50 - 5000微米。还研究了电极宽度对灵敏度(电流强度、电流密度和信噪比)的影响。通过模拟工作电极周围产生的浓度分布来讨论灵敏度。研究了柱填料尺寸和柱尺寸对分离效率的影响。在本研究中,成功实现了三种儿茶素的良好分离,(+)-儿茶素、表儿茶素和表没食子儿茶素没食子酸酯的检测限分别为350、450和160 nM。