Surface modified silver selinide nanoparticles as extracting probes to improve peptide/protein detection via nanoparticles-based liquid phase microextraction coupled with MALDI mass spectrometry

We report the first use of functionalized Ag₂Se nanoparticles (NPs) as effective extracting probes for NPs-based liquid-phase microextraction (NPs-LPME) to analyze hydrophobic peptides and proteins from biological samples (urine and plasma) and soybean in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Surface modified functional groups such as octadecanethiol (ODT) and 11-mercaptoundecanoic acid (MUA) on Ag₂Se NPs were found to play an important role for efficient extraction of peptides and proteins from test samples through hydrophobic interactions. The peptides can be efficiently extracted using functionalized Ag₂Se NPs as extracting probes in the presence of high concentration of matrix interferences such as 4 M urea, 0.5% Triton X-100 and 3% NaCl. Ag₂Se@ODT NPs have shown better extraction efficiency and detection sensitivity for peptides than Ag₂Se@MUA NPs, bare Ag₂Se NPs and conventional MALDI-MS. The LODs are 20-68 nM for valinomycin and 100-180 nM for gramicidin D using Ag₂Se@ODT NPs-LPME in the MALDI-MS. The current approach is highly sensitive and the target analytes can be effectively isolated without sample loss and efficiently analyzed in MALDI-MS.