Development of nanomaterial enabling highly sensitive surface-assisted laser desorption/ionization mass spectrometry peptide analysis.

RATIONALE: Surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) is an approach derived from matrix-assisted laser desorption/ionization (MALDI)-MS which overcomes the drawbacks associated with the use of organic matrices required to co-crystallize with the analytes. Indeed, nanomaterials commonly used in SALDI-MS as inert surfaces to promote desorption/ionization (D/I) ensure straightforward direct deposition of samples while providing mass spectra with ions only related to the compound of interest. The objective of this study was to develop a novel SALDI-MS approach based on steel plates that are surfaces very rapidly and easily tuned to perform the most efficient peptide detection as possible. To compare the SALDI efficacy of such metal substrates, D/I efficiency and deposit homogeneity were evaluated according to steel plate fabrication processes. METHODS: The studied surfaces were nanostructured steel plates that were chemically modified by perfluorosilane and textured according to different frequencies and laser writing powers. The capacity of each tested 100 surfaces was demonstrated by comparative analyses of a mixture of standard peptides (m/z 600-3000) performed with a MALDI-TOF instrument enabling MALDI, SALDI and imaging experiments. RESULTS: A peptide mix was used to screen the different surfaces depending on their D/I efficiency and their ability to ensure homogeneous deposit of the samples. For that purpose, deposition homogeneity was visualized owing to reconstructed ionic images from all protonated or sodiated ions of the 10 peptides constituting the standard mix. CONCLUSIONS: Seven surfaces were then selected satisfying the required D/I efficiency and deposit homogeneity criteria. Results obtained with these optimal surfaces were then compared with those recorded by MALDI-MS analyses used as references.

The Scavenging Effect of Myoglobin from Meat Extracts toward Peroxynitrite Studied with a Flow Injection System Based on Electrochemical Reduction over a Screen-Printed Carbon Electrode Modified with Cobalt Phthalocyanine: Quantification and Kinetics.

The scavenging activity of myoglobin toward peroxynitrite (PON) was studied in meat extracts, using a new developed electrochemical method (based on cobalt phthalocyanine-modified screen-printed carbon electrode, SPCE/CoPc) and calculating kinetic parameters of PON decay (such as half-time and apparent rate constants). As reactive oxygen/nitrogen species (ROS/RNS) affect the food quality, the consumers can be negatively influenced. The discoloration, rancidity, and flavor of meat are altered in the presence of these species, such as PON. Our new highly thermically stable, cost-effective, rapid, and simple electrocatalytical method was combined with a flow injection analysis system to achieve high sensitivity (10.843 nA µM-1) at a nanomolar level LoD (400 nM), within a linear range of 3-180 µM. The proposed biosensor was fully characterized using SEM, FTIR, Raman spectroscopy, Cyclic Voltammetry (CV), Differential Pulse Voltammetry (DPV), and Linear Sweep Voltammetry (LSV). These achievements were obtained due to the CoPc-mediated reduction of PON at very low potentials (around 0.1 V vs. Ag/AgCl pseudoreference). We also proposed a redox mechanism involving two electrons in the reduction of peroxynitrite to nitrite and studied some important interfering species (nitrite, nitrate, hydrogen peroxide, dopamine, ascorbic acid), which showed that our method is highly selective. These features make our work relevant, as it could be further applied to study the kinetics of important oxidative processes in vivo or in vitro, as PON is usually present in the nanomolar or micromolar range in physiological conditions, and our method is sensitive enough to be applied.

Influence of buried oxide layers of nanostructured SOI surfaces on matrix-free LDI-MS performances.

In this paper, we report on the nanostructuration of the silicon crystalline top layer of different "home-made" SOI substrates presenting various buried oxide (BOx) layer thicknesses. The nanostructuration was achieved via a one-step metal assisted chemical etching (MACE) procedure. The etched N-SOI substrate surfaces were then characterized by AFM, SEM and photoluminescence. To investigate their laser desorption/ionization mass spectrometry performances, the different surfaces have been assessed towards peptide mixtures. We have shown that the matrix-free LDI process occurred from surface heating after laser irradiation and was fostered by thermal confinement in the thin nanostructured Si surface layer. This thermal confinement was enhanced with the increase of the buried oxide layer thickness until an optimal thickness of 200 nm for which the best results in terms of signal intensities, peptide discrimination and spot to spot and surface to surface variations were found.

Controlled modification of electrochemical microsystems with polyethylenimine/reduced graphene oxide using electrophoretic deposition: Sensing of dopamine levels in meat samples.

Microsystems play an important role in many biological and environmental applications. The integration of electrical interfaces into such miniaturized systems provides new opportunities for electrochemical sensing where high sensitivity and selectivity towards the analyte are requested. This can be only achieved upon controlled functionalization of the working electrode, a challenge for compact microsystems. In this work, we demonstrate the benefit of electrophoretic deposition (EPD) of reduced graphene oxide/polyethylenimine (rGO/PEI) for the selective modification of a gold (Au) microelectrode in a microsystem comprising a Pt counter and a Ag/AgCl reference electrode. The functionalized microsystem was successfully applied for the sensing of dopamine with a detection limit of 50nM. Additionally, the microsystem exhibited good performance for the detection of dopamine levels in meat samples.

MoS2/TiO2/SiNW surface as an effective substrate for LDI-MS detection of glucose and glutathione in real samples.

Here, we report for the first time, the use of molybdenum disulfide/titanium oxide/silicon nanowires (MoS2/TiO2/SiNW) surfaces for SALDI-MS detection as alternative to MALDI-MS method. Silicon nanowires were fabricated by the well-known metal-assisted chemical etching process followed by the deposition of TiO2 by atomic layer deposition. MoS2 deposition was achieved through hydrothermal treatment. The MoS2/TiO2/SiNW substrate has shown high performance for the detection of small compounds of different molecular weights, including glutathione, glucose, amino acids, antibiotics to name a few. All of the tested compounds, in pure or in mixed solutions were successfully detected in positive ion mode. Therefore, we have also attempted quantitative measurements of GSH and glucose in human blood serum.

Peroxynitrite Sensor Based on a Screen Printed Carbon Electrode Modified with a Poly(2,6-dihydroxynaphthalene) Film.

For the first time the electropolymerization of 2,6-dihydroxynaphthalene (2,6-DHN) on a screen printed carbon electrode (SPCE) was investigated and evaluated for peroxynitrite (PON) detection. Cyclic voltammetry was used to electrodeposit the poly(2,6-DHN) on the carbon electrode surface. The surface morphology and structure of poly(2,6-DHN) film were investigated by SEM and FTIR analysis, and the electrochemical features by cyclic voltammetry. The poly(2,6-DHN)/SPCE sensor showed excellent electrocatalytic activity for PON oxidation in alkaline solutions at very low potentials (0-100 mV vs. Ag/AgCl pseudoreference). An amperometric FIA (flow injection analysis) system based on the developed sensor was optimized for PON measurements and a linear concentration range from 2 to 300 µM PON, with a LOD of 0.2 µM, was achieved. The optimized sensor inserted in the FIA system exhibited good sensitivity (4.12 nA·µM-1), selectivity, stability and intra-/inter-electrode reproducibility for PON determination.