Hydrophilic-interaction chromatography (HILIC) is gaining followers within top-down proteomics scientists. HILIC retention mechanism can help us to better understand glycosylation and its role in several diseases. In this paper, Marta describes a new monolithic material acrylamide-based that outperforms commercially available silica-based HILIC stationary phases at low percentages of trifluoroacetic acid (TFA). In the comparison, our monolithic column showed not only a higher selectivity but also a higher peak capacity than a 2.7-µm-silica-particle column (23 vs 12). Moreover, mass spectrometry detection is also affected positively by a reduced amount of TFA. The signal intensity gained almost one order of magnitude in the analysis of bigger proteins ( > 80 kDa) and the amount of protein-TFA adducts decreased drastically.

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