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Rapid analysis of a single sample could be completed within 10 min. Overall, our results demonstrated that SPME swab-MS is a promising noninvasive method for enhanced detection of analytes in human body.We report capacitively coupled contactless conductivity detection (C4D) of proteins separated by microfluidic capillary isoelectric focusing (μCIEF). To elucidate the evolution of negative conductivity peaks during focusing and seek IEF conditions for sensitive conductivity detection, numerical simulation was performed using a model protein GFP (green fluorescence protein) and hypothetical carrier ampholytes (CAs). C4D was successfully applied to the μCIEF by optimizing assay conditions using a simple and effective pressure-mobilization approach. The conductivity and fluorescence signals of a focused GFP band were co-detected, confirming that the obtained negative C4D peak could be attributed to the actual protein, not the non-uniform background conductivity profile of the focused CAs. GFP concentrations of 10 nM-30 μM was quantified with a detection limit of 10 nM. Finally, the resolving power was analyzed by separating a mixture of R-phycoerythrin (pI 5.01), GFP-F64L (pI 5.48), and RK-GFP (pI 6.02). The conductivities of the three separated fluorescence proteins were measured with average separation resolution of 2.06. We expect the newly developed label-free μCIEF-C4D technique to be widely adopted as a portable, electronics-only protein-analysis tool.MeCP2 is an essential transcriptional repressor that mediates transcriptional inhibition by binding to methylated DNA. The binding specificity of MeCP2 protein to methylated DNA was considered to depend on its methyl-CpG binding domain (MBD). In this study, we used atomic force microscope based single-molecular force spectroscopy to investigate the interaction of MeCP2 MBD and methylated DNA. The specific interaction forces of the MeCP2 MBD-methylated DNA complexes were measured for the first time. The dynamics was also investigated by measuring the unbinding force of the complex at different loading rates. In addition, the distribution of unbinding forces and binding probabilities of MeCP2 MBD and different DNA were studied at the same loading rate. It was found that MeCP2 MBD had weak interaction with hemi-methylated and unmethylated DNA compared to methylated DNA. This work revealed the binding characteristics of MeCP2 MBD and methylated DNA at the single-molecule level. It provides a new idea for exploring the molecular mechanism of MeCP2 in regulating methylation signals.Glucosylsphingosine (GlcS) in plasma is considered to be a reliable biomarker of Gaucher disease. The detection difficulty of GlcS is that it is difficult to achieve simultaneous separation and quantification with its isomer galactosylsphingosine (GalS), a biomarker of Krabbe disease. https://www.selleckchem.com/products/rin1.html In this work, a multiplexed stable isotope labeling absolute quantization strategy coupled with magnetic dispersive solid phase extraction using new prepared dummy magnetic molecularly imprinted polymers (DMMIPs) has been developed for this purpose by ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). 8-Plex Amine-reactive Mass Difference Tags (M360/361/362/363/373/375/376/378-AMDTs), were designed, synthesized and used to label GalS and GlcS in different 8 plasma samples, respectively. Synchronously, M359-AMDTs was prepared and used to label mixed standards of GalS and GlcS, which served as internal standards in UHPLC-MS/MS quantitation. Then DMMIPs possessing dual recognition function were applied for specific enrichment and purification of all GlcS and GalS derivatives from a combined solution of labeled 8-plex plasma samples and mixed standards before UHPLC-MS/MS injection. The labeling efficiency, chromatographic retention and mass spectrometry responses of all the 9 AMDTs reagents were consistent for GlcS and GalS. The established and validated method enabled 8-plex plasma samples quantification in a single UHPLC-MS/MS run ( less then 2.0 min). Good linearity of AMDTs-GlcS/GalS derivatives was obtained in the range of 0.02-800 nM. LODs of GlcS and GalS were both 0.005 nM. The recoveries were in the range of 96.1-107.2%. The method was successfully applied for multiplex quantitative analysis of GlcS and GalS in human plasma samples. The results indicated that this method was capable of better realizing the simultaneous separation and quantification of GalS and GlcS compared to reported methods.In this paper, low-cost stainless steel sheets with excellent electric conductivity were utilized as the robust substrate for fabrication of disposable working electrodes. The stainless steel electrodes were modified with carbon cement and then coupled in paper-based analytical devices for analysis of heavy metals (cadmium and lead) in toys or indole-3-acetic acid (IAA) in plants, respectively. For stripping analysis of cadmium and lead, the dilution ratio of the carbon cement, the pH value of the buffer solution, the pre-deposition potential and time, and the bismuth concentration were optimized with the detection limits reaching 1 μg•L-1. After optimization of the dilution ratio of carbon cement, the similar devices could also be used for analysis of IAA at the concentration of less than 0.5 μM. This strategy could be successfully applied for differentiation of migratable lead in toys or in situ amounts of IAA in root tips of Arabidopsis thaliana in real time, respectively. Our results implied that the electric conductivity of the substrate could possibly be critical for the improvement of the analytical performance of the modified electrodes. This study suggested that stainless steel could become a suitable and cost-effective substrate for fabrication of disposable carbon-based electrodes used in electrochemical detection.Hexafluoroisopropanol-alkanol (C6-C12) supramolecular solvents were proposed and their application potential as restricted access extraction solvents was assessed. Hexafluoroisopropanol acts as alkanol reverse micelle-forming agent and density-regulating agent for the supramolecular solvent synthesis. The formation of supramolecular solvent only needs a small percentage of hexafluoroisopropanol ( less then 10% v/v for alkanol less then 5% v/v) and is almost not affected by the pH (2-11) and low ionic strength (NaCl less then 3%, w/v) of aqueous medium. The supramolecular solvents have higher density than water and consist of reversed micellar aggregates with irregular mesh structures and hydrophilic inner cavities, and the hydrophilic cavity size is obviously dependent on the hexafluoroisopropanol amount in the bulk system but almost not relevant to the alkanol amount. The hydrogen-bond and hydrophobic interactions of hexafluoroisopropanol with alkanol and the hydrogen-bond force between hexafluoroisopropanol and water play crucial roles in the formation of supramolecular solvent.
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