Ultrafast analysis of oligosaccharides on microchip with light-emitting diode confocal fluorescence detection
Ultrafast analysis of oligosaccharides on microchip with light-emitting diode confocal fluorescence detection
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DOI：
10.1002/elps.200390086
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发表时间：
2003-02-01
期刊：
影响因子：
2.9
通讯作者：
Baba, Y
中科院分区：
文献类型：
--
作者：
Dang, F;Zhang, L;Baba, Y
建立了一种基于发光二极管(LED)共聚焦荧光检测的微芯片电泳(MU-CE)快速分离、高灵敏度检测复杂低聚糖的新方法。发现8-氨基比林-1，3，6-三磺酸(APTS)标记的寡糖在聚甲基丙烯酸甲酯(PMMA)微芯片表面有很强的吸附作用。因此，对三类主要的动态涂层添加剂进行了系统的研究，发现纤维素衍生物能够特异性地抑制这种吸附，并允许在PMMA芯片上进行高性能分离。添加剂浓度、缓冲液pH值和外加电场强度是影响APTS标记的寡糖在PMMA芯片上高效分离的关键因素。在最佳条件下，糊精水解液中15种低聚糖在45 S内即可分离，分离效率可达每米400,000个理论塔板以上。测定了14种低聚糖迁移时间的相对标准偏差(RSD)值。APTS标记葡萄糖的检出限为1.98×10~(-8)mol/L或8.61amol，信噪比(S/N)为3。该方法快速、高效、灵敏，可广泛应用于复杂寡糖的分析。
We have developed a new method for the high-speed separation and high-sensitivity detection of complex oligosaccharides based on microchip electrophoresis (mu-CE) with light-emitting diode (LED) confocal fluorescence detection. Oligosaccharides labeled with 8-aminopyrene-1,3,6-trisulfonate (APTS) were found to strongly adsorb to the surface of polymethylmethacrylate (PMMA) microchips. Accordingly, three classes of major dynamic coating additives were systematically investigated, and cellulose derivatives were found to specifically suppress such adsorption and allow high-performance separation on PMMA chips. Additive concentration, buffer pH and applied field strength were found to be key factors in the high-performance separation of APTS-labeled oligosaccharides on PMMA chips. Under optimal conditions, 15 oligosaccharides in dextrin hydrolysate can be separated within 45 s with an electrophoretic separation efficiency of over 400 000 theoretical plates per meter. The relative standard deviation (RSD) values of migration times of fourteen oligosaccharides were. less than 0.50% between six different channels, and the detection limit for APTS-labeled glucose was about 1.98 x 10(-8) mol/L or 8.61 amol with a signal-to-noise ratio (S/N) of 3. The high speed, high efficiency and high sensitivity of this mu-CE-based method indicate that it can be widely applied to analysis of complex oligosaccharides.