Evaluation of commercial micro-flow liquid chromatography columns for mass spectrometry-based proteomics.

This study aimed to evaluate the separation efficiency and loading capacity of four commercially available micro-flow liquid chromatography (micro-flow LC) columns with 1.0 mm i.d. (PepMap™ C18, HALO® ES-C18, YMC-Triart™ C18 and Acquity UPLC Peptide BEH C18, abbreviated as PepMap, HALO, YMC and BEH) for mass spectrometry-based proteomics analysis. Samples including cytochrome c (cyt-c), human plasma, and HeLa protein digest were used for the tests. The YMC showed much wider peak widths than the other columns, exhibited relatively poor identification results. However, the other three columns showed similar identification performance. Among them, the PepMap was the optimal choice for plasma proteomics as it had a high loading capacity and exhibited the most symmetrical peak shape with a symmetry factor closest to 1.0. In general, our results provided valuable and solid support for the choice of a chromatographic column, which could potentially contributes to the wider application of micro-flow LC-MS/MS in proteomics research.