Skelley Alison M, Mathies Richard A
Department of Chemistry, University of California, 307 Lewis Hall, Berkeley, CA 94720, USA.
J Chromatogr A. 2003 Dec 22;1021(1-2):191-9. doi: 10.1016/j.chroma.2003.08.096.
Chiral separations of fluorescamine-labeled amino acids are characterized and optimized on a microfabricated capillary electrophoresis (CE) device. A standard mixture of acidic and neutral amino acids is labeled with fluorescamine in less than 5 min and the hydroxypropyl-beta-cyclodextrin (HPbetaCD) concentration, temperature, and pH are optimized (15 mM HPbetaCD, 6 degrees C, pH < 9) to achieve high-quality and low background chiral separations in less than 200 s. All four stereoisomers formed in the labeling reaction of the chiral dye with the chiral amino acids are typically resolved. At pH > 9, isomerization of the dye chiral center is observed that occurs on the time scale of the chip separation. Typical limits of detection are approximately 50 nM. These results demonstrate the feasibility of combining fluorescamine labeling of amino acids with microfabricated CE devices to develop low-volume, high-sensitivity apparatus and methods for extraterrestrial exploration.
在手性微流控毛细管电泳（CE）装置上对荧光胺标记的氨基酸进行了手性分离表征和优化。酸性和中性氨基酸的标准混合物在不到5分钟内用荧光胺标记，优化了羟丙基-β-环糊精（HPβCD）浓度、温度和pH值（15 mM HPβCD、6℃、pH<9），以在不到200秒的时间内实现高质量、低背景的手性分离。手性染料与手性氨基酸标记反应中形成的所有四种立体异构体通常都能得到分离。在pH>9时，观察到染料手性中心的异构化发生在芯片分离的时间尺度上。典型检测限约为50 nM。这些结果证明了将氨基酸的荧光胺标记与微流控CE装置相结合，开发用于外星探索的小体积、高灵敏度仪器和方法的可行性。
