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Segmented Microfluidics-Based Packing Technology for Chromatographic Columns
- Source :
- Analytical Chemistry. 93:8450-8458
- Publication Year :
- 2021
- Publisher :
- American Chemical Society (ACS), 2021.
-
Abstract
- Nanoflow liquid chromatography-mass spectrometry (NanoLC-MS) has become the method of choice for the analysis of complex biological systems, especially when the available sample amount is limited. The preparation of high-performance capillary columns for nanoLC use is still a technical challenge. Here, we report a segmented microfluidic method for the preparation of packed capillary columns, where liquid segments were used as soft, dynamic, and well-dispersed slurry reservoirs for carrying and delivering micrometer packing particles. Based on this microfluidic packing technology, the column bed was assembled layer-by-layer at a 50 μm resolution, and ultralong capillary columns of 3, 5, and 10 m were fabricated in such a manner. The microfluidically packed columns demonstrated excellent separation efficiencies of 116 000 plates/m. The higher efficiencies obtained at higher slurry concentrations also indicate that a high-quality packed bed can be obtained without sacrificing the packing speed. Kinetic performance limit analysis shows that the microfluidic packed columns have higher peak capacity production efficiency in the high-resolution region, presenting an improved separation impedance of 2800, which is significantly better than columns packed with the conventional slurry packing method. In comparison with a commercial nanoLC column, a 5 m long microfluidic packed column was evaluated for proteomic analysis using a standard HeLa protein digest and presented 261% improvement in peptide identification capability, resulting in significantly enhanced protein identification confidence.
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 93
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....75636755a9d2c66840bf9c62b366c889