Molecular Structure Characterization of Micro/Nanoplastics by 193 nm Ultraviolet Photodissociation Mass Spectrometry.

The degradation of macroplastics results in micro/nanoplastics (MNPs) in the natural environment, inducing high health risks worldwide. It remains challenging to characterize the accurate molecular structures of MNPs. Herein, we integrate 193 nm ultraviolet photodissociation (UVPD) with mass spectrometry to interrogate the molecular structures of poly(ethylene glycol) terephthalate and polyamide (PA) MNPs. The backbones of the MNP polymer can be efficiently dissociated by UVPD, producing rich types of fragment ions. Compared to high-energy collision dissociation (HCD), the structural informative fragment ions and corresponding sequence coverages obtained by UVPD were all improved 2.3 times on average, resulting in almost complete sequence coverage and precise structural interrogation of MNPs. We successfully determine the backbone connectivity differences of MNP analogues PA6, PA66, and PA610 by improving the average sequence coverage from 26.8% by HCD to 89.4% by UVPD. Our results highlight the potential of UVPD in characterizing and discriminating backbone connectivity and chain end structures of different types of MNPs.

Probing conformational hotspots for the recognition and intervention of protein complexes by lysine reactivity profiling.

Probing the conformational and functional hotspot sites within aqueous native protein complexes is still a challenging task. Herein, a mass spectrometry (MS)-based two-step isotope labeling-lysine reactivity profiling (TILLRP) strategy is developed to quantify the reactivities of lysine residues and probe the molecular details of protein-protein interactions as well as evaluate the conformational interventions by small-molecule active compounds. The hotspot lysine sites that are crucial to the SARS-CoV-2 S1-ACE2 combination could be successfully probed, such as S1 Lys417 and Lys444. Significant alteration of the reactivities of lysine residues at the interaction interface of S1-RBD Lys386-Lys462 was observed during the formation of complexes, which might be utilized as indicators for investigating the S1-ACE2 dynamic recognition and intervention at the molecular level in high throughput.

Salt-effect enhanced hollow-fiber liquid-phase microextraction of glutathione in human saliva followed by miniaturized capillary electrophoresis with amperometric detection.

A hollow-fiber liquid-phase microextraction (HF-LPME) method was established for purification and enrichment of glutathione (GSH) in human saliva followed by a miniaturized capillary electrophoresis with amperometric detection system (mini-CE-AD). Based on regulating isoelectric point and increasing salt effect to modify donor phase, HF-LPME could provide high enrichment efficiency for GSH up to 471 times, and the extract was directly injected for mini-CE-AD analysis. The salt-effect enhanced HF-LPME/mini-CE-AD method has been successfully applied to saliva analysis, and acceptable LOD (0.46 ng/mL, S/N = 3) and recoveries (92.7-101.3%) could be obtained in saliva matrix. The sample pretreatment of this developed method was simple and required no derivatization, providing a potential alternative for non-invasive fluid analysis using portable instrument.