A microdevice with integrated liquid junction for facile peptide and protein analysis by capillary electrophoresis/electrospray mass spectrometry.
A microdevice with integrated liquid junction for facile peptide and protein analysis by capillary electrophoresis/electrospray mass spectrometry.
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DOI：
10.1021/ac991150z
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发表时间：
2000-01
影响因子：
7.4
通讯作者：
B. Zhang;F. Foret;B. Karger
中科院分区：
文献类型：
--
作者：
B. Zhang;F. Foret;B. Karger
设计了一种毛细管电泳与质谱（CE/MS）快速耦合的微型装置。该器件由玻璃晶圆制成，采用标准光刻/湿化学蚀刻方法。该设计集成了(a)样品入口，(b)分离通道，(c)液体结，以及(d)用于插入电喷雾毛细管的引导通道，该通道被封闭在离子阱ms的小型化亚大气电喷雾室中。可替换的电喷雾毛细管通过微制造的引导通道精确地与分离通道的出口对齐。不需要胶水来密封电喷雾毛细管。这种设计可以简单快速地更换微型装置或电喷雾毛细管。对该装置的性能进行了多肽、蛋白质和蛋白质胰蛋白酶消化的CE/MS测试。采用在线串联质谱法对蛋白质消化产物进行结构鉴定。在较长的通道（片上11 cm）上可以获得高效/高分辨率的分离，而在较短的通道（片上4.5 cm）上可以实现快速分离（50 s以下）。实验中采用了电动注射和压力注射两种方法。分离效率与传统毛细管电泳相当。
A novel microfabricated device was implemented for facile coupling of capillary electrophoresis with mass spectrometry (CE/MS). The device was constructed from glass wafers using standard photolithographic/wet chemical etching methods. The design integrated (a) sample inlet ports, (b) the separation channel, (c) a liquid junction, and (d) a guiding channel for the insertion of the electrospray capillary, which was enclosed in a miniaturized subatmospheric electrospray chamber of an ion trap MS. The replaceable electrospray capillary was precisely aligned with the exit of the separation channel by a microfabricated guiding channel. No glue was necessary to seal the electrospray capillary. This design allowed simple and fast replacement of either the microdevice or the electrospray capillary. The performance of the device was tested for CE/MS of peptides, proteins, and protein tryptic digests. On-line tandem mass spectrometry was used for the structure identification of the protein digest products. High-efficiency/high-resolution separations could be obtained on a longer channel (11 cm on-chip) microdevice, and fast separations (under 50 s) were achieved with a short (4.5 cm on-chip) separation channel. In the experiments, both electrokinetic and pressure injections were used. The separation efficiency was comparable to that obtained from conventional capillary electrophoresis.