Ethane-bridged hybrid monoliths with well-defined mesoporosity and great stability for high-performance peptide separation

A novel kind of hybrid monolith with well-defined mesopore structure was prepared based on sol-gel condensation of 1, 2-bis(trimethoxysilyl) ethane (BTME) and tetramethoxysilane. Compared with terminal organosiloxanes used for preparation of conventional hybrid monoliths, BTME, as an ethanebridged alkoxysilane precursor, could not only maintain the uniformity of pore size distribution, but also improve the chemical stability of the monolith via Si-C bonds in the framework. Owing to the controllable mesoporous structure and good stability, the monolithic column was used for the separation of peptides with half peak width less than 6 s and the run-to-run and column-to-column relative standard deviations (RSD) for the retention time of five standard peptides less than 2.5%, showing narrow peak width and good reproducibility. Moreover, the separation performance could be well maintained even after washed by the mobile phase with pH 11.0 at 50 degrees C. Furthermore, 100 cm-length monolithic column was prepared and successfully used for nanoRPLC-ESI-MS/MS analysis of HeLa cell lysate digests, and 5670 proteins corresponding to 37574 peptides were identified from 750 ng of the sample, showing great promising of single-shot large-scale in-depth proteomic research. (c) 2018 Elsevier B.V. All rights reserved.