On-Chip Comparison of the Performance of First- and Second-Generation Micropillar Array Columns

Because of its dimensions, the recently introduced micropillar array columns are most suited for high-efficiency liquid chromatography separations in proteomics. Unlike the packed bed columns and capillary-based column formats, the micropillar array concept still has significant room to progress in terms of the reduction of its characteristic size (i.e., pillar diameter and interpillar distance) to open the road to even higher-efficiency separations and their applications. We report here on the on-chip comparison between first-generation (Gen 1) and second-generation (Gen 2) micropillar array columns wherein the pillar and interpillar size have been halved. Because of the on-chip measurements, the observed plate heights H represent the fundamental band broadening, devoid of any extra-column band-broadening effects. The observed reduction of H with a factor of 2 around the u opt-velocity and with a factor of 4 in the C-term dominated regime of the van Deemter-curve is in full agreement with the theoretically expected gain. This shows the pillar and interpillar size reduction could be effectuated without affecting the theoretical separation potential of the micropillar arrays. Compared to Gen 1, Gen 2 offers a 4-fold reduction of the required analysis time around the optimal velocity and about a 16-fold reduction in the C-term-dominated range.

Automated summary

Due to their dimensions, micropillar array columns are ideal for high-efficiency liquid chromatography separations in proteomics. The micropillar array concept has room to progress in terms of size reduction, which can lead to even higher-efficiency separations and applications. A comparison between first-generation (Gen 1) and second-generation (Gen 2) micropillar array columns on a chip showed that reducing the pillar and interpillar size did not affect the theoretical separation potential, resulting in significant reductions in analysis time.