Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
Anal Chem. 2010 Feb 15;82(4):1420-6. doi: 10.1021/ac902491x.
In this study, we show for the first time that the separation efficiency of a pillar array column under pressure-driven liquid chromatography (LC) conditions can be improved using a separation channel with low-dispersion turns. The pillar array column was fabricated by reactive ion etching of a silicon substrate. With the low-dispersion-turn geometry, a column with a length and width of 110 mm and 400 microm, respectively, could be fabricated on a 20 x 20 mm microchip. Under nonretained conditions, the solute bands obtained for fluorescent compounds remained almost unchanged even after passing through the low-dispersion turns; however, significant skewing of the solute bands was observed in the case of constant-radius turns. Two coumarin dyes were well resolved under reversed-phase conditions, and a maximum theoretical plate number of 8000 was obtained. Successful separation of the fluorescent derivatives of six amino acids was achieved in 140 s. These results indicated that the separation efficiency of microchip chromatography could be significantly improved using a long separation channel with low-dispersion turns.
在这项研究中,我们首次表明,在压力驱动的液相色谱(LC)条件下,使用具有低分散转弯的分离通道可以提高支柱阵列柱的分离效率。支柱阵列柱是通过硅基板的反应离子刻蚀制造的。使用低分散转弯几何形状,可以在 20 x 20 毫米的微芯片上制造长度和宽度分别为 110 毫米和 400 微米的柱。在无保留条件下,即使经过低分散转弯,荧光化合物的溶质带也几乎保持不变; 然而,在等半径转弯的情况下,观察到溶质带的明显偏斜。在反相条件下,两种香豆素染料得到了很好的分离,得到了最大的理论板数为 8000。在 140 秒内成功分离了六种氨基酸的荧光衍生物。这些结果表明,使用具有低分散转弯的长分离通道可以显著提高微芯片色谱的分离效率。