Nano‐liquid chromatography with a new nano‐structured monolithic nanocolumn for proteomics analysis

Abstract

Herein, we report the preparation and application of a new nano-structured monolithic nanocolumn based on modified graphene oxide using narrow fused silica capillary column (e.g., 50 μm internal diameter). The nanocolumn was prepared by an in situ polymerization using butyl methacrylate, ethylene dimethacrylate, and methacryloyl graphene oxide nanoparticles. Dimethyl formamide and water were used as the porogenic solvent. After polymerization, the obtained nanocolumn was coated with dimethyloctadecylchlorosilane in order to enhance the hydrophobicity. Both isocratic and gradient nano-liquid chromatographic separations for small molecules (e.g., alkylbenzenes) and macromolecules (e.g., intact proteins) were performed. Theoretical plates number up to 3600 plates/m in isocratic mode for propylbenzene were achieved. It was demonstrated that the feasibility of graphene oxide modified monolithic nanocolumn for high-efficiency and high-throughput nanoscale proteomics analysis. The high resolving power of monolithic nanocolumn yielded sensitive protein separation with narrower peak width while a high-resolution analysis of peptides from trypsin-digested cytochrome C could be obtained. Graphene oxide based monolithic nanocolumns are promising and can allow to powerful tools for trace proteom sample analysis.

Hydrophobic AEROSIL®R972 Fumed Silica Nanoparticles Incorporated Monolithic Nano-Columns for Small Molecule and Protein Separation by Nano-Liquid Chromatography

“…In a very recent research report, BMA-based monolithic columns were prepared and applied for proteomics analysis by nano-LC [ 24 ]. Although this column demonstrated satisfactory performance, the polymerization content needs to be optimized due to new nanostructures (e.g., HFSNPs).…”

“…The composition of the polymerization solutions used in the preparation of the monoliths is given in Table 1 . The optimization of BMA and EDMA contents was achieved according to the published literature [ 24 ]. The selection of the solvent system was the first critical issue for both the solubility and the dispersion of HFSNPs into the polymerization solution, which is essential for the preparation of HFSNPs incorporated monoliths.…”

Determination of the geographic origin of 52 honey samples based on the assessment of anionic content profiling with a new algorithm using monolithic column‐based micellar nano‐liquid chromatography

“…Various monolithic columns were prepared and applied for the analysis of anionic content (Aydoğan, 2015; Lin et al., 2019). In our recent study, a new BMA‐based nano‐structured monolithic column with 50 µm ID was prepared and applied for proteomics analysis (Günyel et al., 2021). In this study, the same monolithic column with 100 µm ID was prepared and used for the analysis of anions in 52 honey samples.…”

“…The same procedure for the column preparation was just applied according to the published article (Günyel et al., 2021). Briefly, graphite was used to obtain GO and then, methacryloly graphene oxide nanoparticles (MGNOPs) were synthesized (Aydoğan et al., 2021).…”

Fabrication of monolithic capillary columns with inner diameter 50–530 μm employing a mixture of pentaerythritol tetraacrylate and polyhedral oligomeric silsesquioxane‐methacrylate as crosslinkers

“…An attachment of nanoparticles to the monolithic surface of the stationary phase is yet another way how to improve the chromatographic performance of polymer monoliths [28,29]. However, this approach is not used so often as the direct incorporation of nanoparticles into the monolith structure where monomers containing, for example, C60 fullerene, silica, or graphene oxide nanoparticles are utilized for the synthesis of monolithic columns [30][31][32][33].…”