Capillary Flow LC-MS using µPAC™ columns-Combining Sensitivity with Robustness and Throughput
Mass-spectrometry based proteomics has become an essential tool in biological, biomedical and biopharmaceutical research. Due to the increased sensitivity that can be achieved compared to analytical flow LC-MS, the majority of LC-MS based proteomics research is performed within the nanoflow LC regime, with flow rates typically below 1 μL/min and total run times exceeding 60 min. Compared to the LC-MS methods employed in other ‘omics’ fields, the need for high separation power often prevails over sample throughput, making relatively long separations the gold standard In LC-MS based proteomics. Capillary and microflow LC-MS solutions are recently gaining interest as the demand for large quantitative proteomics studies that require increased throughput and robustness is growing. By working at higher flow rates, the impact of gradient delay and sample loading volumes on the total analysis time can be reduced and low flow rate or column dimension related technical errors (such as electrospray instability, column clogging or the presence of void volumes in the analytical flow path) can be minimized.
A micro Pillar Array Column based solution for capillary flow LC-MS is presented. In contrast to conventional LC columns that contain randomly packed beads as their stationary phase, micro-chip based pillar array chromatography columns have a separation bed of perfectly ordered and free-standing pillars obtained by lithographic etching of a silicon wafer. The regular mobile phase flow pattern through these micro-chip pillar array columns adds very little dispersion to the overall separation, resulting in better peak resolution, sharper elution and increased sensitivity. The free-standing nature of the pillars also leads to much lower back pressure buildup, and makes it possible to operate longer columns.
In this webinar, Dr. Geert van Raemdonck and Robert van Ling of PharmaFluidics explained the principles of the micro-chip based pillar array columns and demonstrated how the PharmaFluidics µPAC™ capLC column can contribute to LC-MS workflow robustness and showcased high resolution data.
Watch it here!