200 μm ID Chip Based Columns for Increased Throughput Peptide Quantitation with NanoLC/MS (original) (raw)

Journal of biomolecular techniques, 2012

Abstract

Nanoflow liquid chromatography coupled with mass spectrometry (nanoLC/MS) is the method of choice for sensitive peptide quantitation. We recently reported on a new microfluidic platform (cHiPLC-nanoflex) for nanoLC-MS applications. In addition to delivering easy-to-use, dead-volume-free connections to microfluidic devices, the platform's cHiPLC columns provide excellent column-to-column separation reproducibility. While providing excellent sensitivity, the low flow rates when using 75 μm ID columns reduces sample throughput due to gradient delay in the nanoLC system itself, delay in the autosampler and sample loop, and the delay caused by the connecting tubing. One way to address this is to use a larger inner diameter column at a proportionally higher flow rate. While this will reduce sensitivity when the total sample amount is kept equal, the delay times, however, can be greatly reduced and sample throughput accelerated. Reducing the column and gradient lengths can further redu...

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