A "plug-and-use" approach towards facile fabrication of capillary columns for high performance nanoflow liquid chromatography.
A "plug-and-use" approach towards facile fabrication of capillary columns for high performance nanoflow liquid chromatography.
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DOI：
10.1016/j.chroma.2013.12.002
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发表时间：
2014-01
期刊：
影响因子：
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通讯作者：
Zhiliang Xiao;Lin Wang;Ya Liu;Qiuquan Wang;Bo Zhang
中科院分区：
文献类型：
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作者：
Zhiliang Xiao;Lin Wang;Ya Liu;Qiuquan Wang;Bo Zhang
毛细管柱用于纳米流液相色谱在现代蛋白质组学研究中发挥着重要作用。需要高质量的色谱柱来提供高峰容量和高度可重复的分离。当在寻找潜在的生物标志物中平行比较多个分离时，这是极其重要的。在此，我们介绍了一种“即插即用”的烧结技术，用于制造高质量和高重现性的毛细管柱。由于采用的预制玻璃料长度相同、渗透性好、稳定性好，毛细管柱在柱间保留时间、峰宽和峰容量方面表现出良好的性能一致性（相对标准偏差，RSD，分别为0.4- 0.9%，2.1- 3.6%和2.7%，n= 6），用于分离蛋白质消化物的复杂混合物。对于填充5 μm颗粒的毛细管柱，最小塔板高度为11 μm，接近该材料的理论性能极限，表现出较高的分离效率。对于分离蛋白质酶，柱表现出优异的峰容量为110和300，分别为20和360分钟的梯度。这种柱的简单制造、良好性能以及一致的质量为需要并行使用多个高性能毛细管柱的高通量分离提供了可靠的工具。
Capillary columns used for nanoflow liquid chromatography play an important role in modern proteomics. High quality columns are needed to provide high peak capacity and highly reproducible separations. This is extremely important when multiple separations were compared in parallel in searching for potential biomarkers. Herein, we introduce a “plug-and-use” fritting technology for fabrication of high quality and highly reproducible capillary columns. Due to the identical length, good permeability, and stability of the prefabricated frits adopted, the capillary columns presented excellent performance consistency in terms of retention time, peak width as well as peak capacity at a column-to-column level (relative standard deviations, RSDs, at 0.4–0.9%, 2.1–3.6%, and 2.7%, respectively,n= 6) for separations of complex mixtures of protein digest. For capillary columns packed with 5 μm particles, high separation efficiency was demonstrated by the minimum plate height of 11 μm, approaching the theoretical performance limit of such material. For separations of protein digests, the columns demonstrated excellent peak capacities of 110 and 300 for 20 and 360 min gradients, respectively. The simple fabrication, good performance as well as consistent quality of such columns provide a reliable tool for high throughput separations requiring the use of multiple high performance capillary columns in parallel.