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Intracellular environment includes proteins, sugars, and nucleic acids interacting in restricted media. In the cytoplasm, the excluded volume effect takes up to 40% of the volume available for occupation by macromolecules. In this work, we tested several approaches modeling crowded solutions for protein diffusion. We experimentally showed how the protein diffusion deviates from conventional Brownian motion in artificial conditions modeling the alteration of medium viscosity and rigid spatial obstacles. The studied tracer proteins were globular bovine serum albumin and intrinsically disordered α-casein. Using the pulsed field gradient NMR, we investigated the translational diffusion of protein probes of different structures in homogeneous (glycerol) and heterogeneous (PEG 300/PEG 6000/PEG 40,000) solutions as a function of crowder concentration. Our results showed fundamentally different effects of homogeneous and heterogeneous crowded environments on protein self-diffusion. In addition, the applied "tracer on lattice" model showed that smaller crowding obstacles (PEG 300 and PEG 6000) create a dense net of restrictions noticeably hindering diffusing protein probes, whereas the large-sized PEG 40,000 creates a "less restricted" environment for the diffusive motion of protein molecules.
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Caseínas , Soroalbumina Bovina , Caseínas/química , Movimento (Física) , DifusãoRESUMO
Aiming toward the production and characterization of delicious and functional gel foods, this communication studies the flavor release from cinnamon-containing κ-carrageenan gel. Cinnamaldehyde, which provides the flavor of cinnamon, was released in a trace amount from the gel and detected by flame ionization detector gas chromatography. The retention of cinnamaldehyde in κ-carrageenan gel and the interaction between flavor and polysaccharide were investigated by high-resolution magic-angle spinning nuclear magnetic resonance (HR-MAS NMR) and pulsed-field gradient NMR (PFG NMR). The intact cinnamaldehyde in the gel was also observed by HR-MAS NMR. The relative mobility difference of the flavor and polysaccharide molecules was observed from the diffusion-ordered NMR spectrum of PFG NMR.
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The formulation of drug/polymer amorphous solid dispersions (ASDs) is one of the most successful strategies for improving the oral bioavailability of poorly soluble active pharmaceutical ingredients (APIs). Hot-melt extrusion (HME) is one method for preparing ASDs that is growing in importance in the pharmaceutical industry, but there are still substantial gaps in our understanding regarding the dynamics of drug dissolution and dispersion in viscous polymers and the physical stability of the final formulations. Furthermore, computational models have been built to predict optimal processing conditions, but they are limited by the lack of experimental data for key mass transport parameters, such as the diffusion coefficient. The work presented here reports direct measurements of API diffusion in pharmaceutical polymer melts, using high-temperature pulsed-field gradient NMR. The diffusion coefficient of a model drug/polymer system (paracetamol/copovidone) was determined for different drug loadings and at temperatures relevant to the HME process. The mechanisms of the diffusion process are also explored with the Stokes-Einstein and Arrhenius models. The results show that diffusivity is linked exponentially to temperature. Furthermore, this study includes rheological characterization, differential scanning calorimetry (DSC), and 1H ssNMR T1 and T1ρ measurements to give additional insights into the physical state, phase separation, and API/polymer interactions in paracetamol/copovidone ASD formulations.
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Acetaminofen/química , Composição de Medicamentos/métodos , Pirrolidinas/química , Compostos de Vinila/química , Espectroscopia de Ressonância Magnética , Polímeros/químicaRESUMO
Self-diffusivities of ethane were measured by multinuclear pulsed field gradient (PFG) NMR inside zeolitic imidazolate framework-11 (ZIF-11) crystals dispersed in several selected polymers to form mixed-matrix membranes (MMMs). These diffusivities were compared with the corresponding intracrystalline self-diffusivities in ZIF-11 crystal beds. It was observed that the confinement of ZIF-11 crystals in ZIF-11 / Torlon MMM can lead to a decrease in the ethane intracrystalline self-diffusivity. Such diffusivity decrease was observed at different temperatures used in this work. PFG NMR measurements of the temperature dependence of the intracrystalline self-diffusivity of ethylene in the same ZIF-11 / Torlon MMM revealed similar diffusivity decrease as well as an increase in the diffusion activation energy in comparison to those in unconfined ZIF-11 crystals in a crystal bed. These observations for ethane and ethylene were attributed to the reduction of the flexibility of the ZIF-11 framework due to the confinement in Torlon leading to a smaller effective aperture size of ZIF-11 crystals. Surprisingly, the intra-ZIF diffusion selectivity for ethane and ethylene was not changed appreciably by the confinement of ZIF-11 crystals in Torlon in comparison to the selectivity in a bed of ZIF-11 crystals. No ZIF-11 confinement effects leading to a reduction in the intracrystalline self-diffusivity of ethane and ethylene were observed for the other two studied MMM systems: ZIF-11 / Matrimid and ZIF-11 / 6FDA-DAM. The absence of the confinement effect in the latter MMMs can be related to the lower values of the polymer bulk modulus in these MMMs in comparison to that in ZIF-11 / Torlon MMM. In addition, there may be a contribution from possible differences in the ZIF-11/polymer adhesion in different MMM types.
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Electrophoretic nuclear magnetic resonance (eNMR) is a powerful tool in studies of nonaqueous electrolytes, such as ionic liquids. It delivers electrophoretic mobilities of the ionic constituents and thus sheds light on ion correlations. In applications of liquid electrolytes, uncharged additives are often employed, detectable via 1 H NMR. Characterizing their mobility and coordination to charged entities is desirable; however, it is often hampered by small intensities and 1 H signals overlapping with major constituents of the electrolyte. In this work, we evaluate methods of phase analysis of overlapping resonances to yield electrophoretic mobilities even for minor constituents. We use phase-sensitive spectral deconvolution via a set of Lorentz distributions for the investigation of the migration behavior of additives in two different ionic liquid-based lithium salt electrolytes. For vinylene carbonate as an additive, no field-induced drift is observed; thus, its coordination to the Li+ ion does not induce a correlated drift with Li+ . On the other hand, in a solvate ionic liquid with tetraglyme (G4) as an additive, a correlated migration of tetraglyme with lithium as a complex solvate cation is directly proven by eNMR. The phase evaluation procedure of superimposed resonances thus broadens the applicability of eNMR to application-relevant complex electrolyte mixtures containing neutral additives with superimposed resonances.
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ZSM-5 zeolite nanoboxes with accessible meso-micro-pore architecture and strong acid sites are important in relevant heterogeneous catalysis suffering from mass transfer limitations and weak acidities. Rational design of parent zeolites with concentrated and non-protective coordination of Al species can facilitate post-synthetic treatment to produce mesoporous ZSM-5 nanoboxes. In this work, a simple and effective method was developed to convert parent MFI zeolites with tetrahedral extra-framework Al into Al-enriched mesoporous ZSM-5 nanoboxes with low silicon-to-aluminium ratios of ≈16. The parent MFI zeolite was prepared by rapid ageing of the zeolite sol gel synthesis mixture. The accessibility to the meso-micro-porous intra-crystalline network was probed systematically by comparative pulsed field gradient nuclear magnetic resonance diffusion measurements, which, together with the strong acidity of nanoboxes, provided superb catalytic activity and longevity in hydrocarbon cracking for propylene production.
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The understanding of major zeolite applications is partially based on diffusion of molecules inside or outside microporous networks. However, it is still a challenge to measure such phenomena. The diffusion ordered nuclear magnetic resonance spectroscopy (DOSY) technique has been reported to measure a probe molecule's diffusion inside porous solids. Pulsed-field gradient (PFG)-NMR has been used herein to measure the self-diffusivity of different probe molecules, such as neopentane, benzene, toluene and 1-dodecene with increasing dynamic diameter, respectively, on a series of H-ZSM-5 zeolites. The latter materials exhibit different crystal sizes, Si/Al ratios and the presence (or absence) of crystalline defects. In addition, shaped zeolite bodies representing industrial catalysts were compared with the afore-mentioned samples.
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Self-diffusion of pure gases including carbon dioxide, methane, ethylene, ethane, and xenon as well as selected two-component mixtures was studied in hybrid zeolitic imidazolate framework-7-8 (ZIF-7-8) crystals using pulsed field gradient (PFG) NMR. This material was formed by mixing 2-methylimidazolate (ZIF-8 linker) and bulkier benzimidazolate (ZIF-7 linker) in the same framework. The intracrystalline diffusion data measured in mixed-linker ZIF-7-8 was compared with the corresponding data in the parent ZIF-8 material. It was found that under the same or comparable experimental conditions the intracrystalline gas diffusion was always slower in ZIF-7-8 than in ZIF-8. This observation is consistent with the expected lower pore aperture size in ZIF-7-8 than in ZIF-8. At the same time, the ethane/ethylene diffusion selectivity was found to be similar in both ZIFs. It was also observed that for the pure studied gases larger than carbon dioxide the diffusivity ratios in ZIF-8 and ZIF-7-8 do not increase with increasing gas size at all loading pressures used. All these data are attributed to greater framework flexibility effects in ZIF-7-8 than ZIF-8. Such effects manifest themselves in a distortion and/or increase in the aperture size in the presence of large sorbates due to linker flexibility.
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Oil is a prominent, but multifaceted material class with a wide variety of applications. Technical oils, crude oils as well as edibles are main subclasses. In this review, the question is addressed how low-field NMR can contribute in oil characterization as an analytical tool, mainly with respect to quality control. Prerequisite in the development of a quality control application, however, is a detailed understanding of the oils and of the measurement. Low-field NMR is known as a rich methodical toolbox that was and is explored and further developed to address questions about oils, their quality, and usability as raw materials, during production and formulation as well as in use.
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Developing high-performance all-solid-state batteries is contingent on finding solid electrolyte materials with high ionic conductivity and ductility. Here we report new halide-rich solid solution phases in the argyrodite Li6 PS5 Cl family, Li6-x PS5-x Cl1+x , and combine electrochemical impedance spectroscopy, neutron diffraction, and 7 Li NMR MAS and PFG spectroscopy to show that increasing the Cl- /S2- ratio has a systematic, and remarkable impact on Li-ion diffusivity in the lattice. The phase at the limit of the solid solution regime, Li5.5 PS4.5 Cl1.5 , exhibits a cold-pressed conductivity of 9.4±0.1â mS cm-1 at 298â K (and 12.0±0.2â mS cm-1 on sintering)-almost four-fold greater than Li6 PS5 Cl under identical processing conditions and comparable to metastable superionic Li7 P3 S11 . Weakened interactions between the mobile Li-ions and surrounding framework anions incurred by substitution of divalent S2- for monovalent Cl- play a major role in enhancing Li+ -ion diffusivity, along with increased site disorder and a higher lithium vacancy population.
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Pulsed-field gradient nuclear magnetic resonance has seen an increase in applications spanning a broad range of disciplines where molecular translational diffusion properties are of interest. The current study introduces and experimentally evaluates the measurement of translational diffusion coefficients of 15N-enriched biomolecules using a 1H-15N HMQC-filtered band-selective excitation short transient (BEST) sequence as an alternative to the previously described SOFAST-XSTE sequence. The results demonstrate that accurate translational diffusion coefficients of 15N-labelled peptides and proteins can be obtained using this alternative 1H-15N HMQC-filtered BEST sequence which is implementable on NMR spectrometers equipped with probes fitted with a single-axis field gradient, including most cryoprobes dedicated to bio-NMR. The sequence is of potential use for direct quantification of protein or peptide translational diffusion within complex systems, such as in mixtures of macromolecules, crowded solutions, membrane-mimicking media and in bicontinuous cubic phases, where conventional sequences may not be readily applicable due to the presence of intense signals arising from sources other than the protein or peptide under investigation.
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Ressonância Magnética Nuclear Biomolecular , Sequência de Aminoácidos , Difusão , Fosfopeptídeos/química , Dodecilsulfato de Sódio/química , SoluçõesRESUMO
To gain new insights into the interaction of proteins and disaccharides, we investigated the hydrodynamic radii, RhProt, of lysozyme molecules in solution and in a ternary protein-sugar-water system by PFG-NMR. Our approach is based on the assumption that the anhydrobiotic properties of disaccharides like trehalose are based on aggregation of sugar molecules to the proteins, i.e., accumulation of sugar molecules close to the protein, and that this process can be investigated by the experimentally detectable RhProt value of the protein. The Rh values are calculated from the experimentally determined diffusion coefficients and the application of a viscosity correction using the inert molecule dioxane as an internal viscosity reference. The experiments were performed as a function of sugar concentration, the overall particle concentration and the pH value. We investigated the disaccharides trehalose and sucrose, mainly for the reason that trehalose has well know cryptobiotic properties while sucrose, which is similar in size and structure, lacks these properties. The results show the formation of a protective sugar shell around the proteins over a wider range of concentrations and pH values in the case of trehalose.
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Muramidase/metabolismo , Sacarose/metabolismo , Trealose/metabolismo , Difusão , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Muramidase/química , Ligação Proteica , Sacarose/química , Trealose/química , ViscosidadeRESUMO
Mass transfer in biofilms is determined by diffusion. Different mostly invasive approaches have been used to measure diffusion coefficients in biofilms, however, data on heterogeneous biomass under realistic conditions is still missing. To non-invasively elucidate fluid-structure interactions in complex multispecies biofilms pulsed field gradient-nuclear magnetic resonance (PFG-NMR) was applied to measure the water diffusion in five different types of biomass aggregates: one type of sludge flocs, two types of biofilm, and two types of granules. Data analysis is an important issue when measuring heterogeneous systems and is shown to significantly influence the interpretation and understanding of water diffusion. With respect to numerical reproducibility and physico-chemical interpretation, different data processing methods were explored: (bi)-exponential data analysis and the Γ distribution model. Furthermore, the diffusion coefficient distribution in relation to relaxation was studied by D-T2 maps obtained by 2D inverse Laplace transform (2D ILT). The results show that the effective diffusion coefficients for all biofilm samples ranged from 0.36 to 0.96 relative to that of water. NMR diffusion was linked to biofilm structure (e.g., biomass density, organic and inorganic matter) as observed by magnetic resonance imaging and to traditional biofilm parameters: diffusion was most restricted in granules with compact structures, and fast diffusion was found in heterotrophic biofilms with fluffy structures. The effective diffusion coefficients in the biomass were found to be broadly distributed because of internal biomass heterogeneities, such as gas bubbles, precipitates, and locally changing biofilm densities. Thus, estimations based on biofilm bulk properties in multispecies systems can be overestimated and mean diffusion coefficients might not be sufficiently informative to describe mass transport in biofilms and the near bulk.
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Bactérias/química , Biofilmes , Espectroscopia de Ressonância Magnética/métodos , Modelos Biológicos , Modelos Químicos , Água/química , Absorção Fisiológica , Simulação por Computador , DifusãoRESUMO
Magnetic impurities are ubiquitous in natural porous media such as sand and soil. They generate internal magnetic field gradients because of increased magnetic susceptibility differences between solid and liquid phase in the pore space and because of the presence of magnetic centers. These internal gradients accelerate NMR relaxation rates and thus might limit the possibility of pore space characterization using NMR. In this study, we investigate the effects of coating the surface of natural sands by the antiferromagnetic iron oxyhydroxide goethite on NMR relaxation and diffusion properties. We found a non-quadratic dependence of the relaxation time distributions on the echo time indicating that the relaxation experiments were not performed in the fast diffusion limit, while the weak dependence on the external magnetic field strength is explained by the preponderance of the surface relaxation over the effect of diffusion in internal gradients. The surface to volume ratio of the pore space, determined by NMR diffusimetry ((S/V)NMR ) remains approximately constant, whereas the same quantity, determined from gas adsorption ((S/V)BET ) increases proportional to the coating density. This is because gas adsorption measures surface roughness on sub-nanometer scale, whereas NMR diffusimetry averages over structures smaller than few microns. This has consequences for the calculation of the surface relaxivities. The usage of the (S/V)NMR leads to constant values, whereas the usage of (S/V)BET leads to apparently decreasing relaxivities with increasing coating, which is unrealistic. Copyright © 2016 John Wiley & Sons, Ltd.
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Espectroscopia de Ressonância Magnética/métodos , Quartzo/química , Dióxido de Silício/química , Algoritmos , Difusão , Campos Eletromagnéticos , Gases , Sedimentos Geológicos , Compostos de Ferro , Minerais , PorosidadeRESUMO
This is a comparative study to clarify the effect of the introduction of layered double hydroxide (LDH) into various polymer matrices. One perfluorosulfonic acid polymer, i.e., Nafion, and two polyaromatic polymers such as sulfonated polyether ether ketone (sPEEK) and sulfonated polysulfone (sPSU), were used for the preparation of nanocomposite membranes at 3 wt.% of LDH loading. Thereafter, the PEMs were characterized by X-ray diffraction (XRD) and dynamic mechanical analysis (DMA) for their microstructural and thermomechanical features, whereas water dynamics and proton conductivity were investigated by nuclear magnetic resonance (PFG and T1) and EIS spectroscopies, respectively. Depending on the hosting matrix, the LDHs can simply provide additional hydrophilic sites or act as physical crosslinkers. In the latter case, an impressive enhancement of both dimensional stability and electrochemical performance was observed. While pristine sPSU exhibited the lowest proton conductivity, the sPSU/LDH nanocomposite was able to compete with Nafion, yielding a conductivity of 122 mS cm-1 at 120 °C and 90% RH with an activation energy of only 8.7 kJ mol-1. The outcome must be ascribed to the mutual and beneficial interaction of the LDH nanoplatelets with the functional groups of sPSU, therefore the choice of the appropriate filler is pivotal for the preparation of highly-performing composites.
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The development of nanocomposite membranes based on hydrocarbon polymers is emerging as one of the most promising strategies for overcoming the performance, cost, and safety limitations of Nafion, which is the current benchmark in proton exchange membranes for fuel cell applications. Among the various nanocomposite membranes, those based on sulfonated polysulfone (sPSU) and Layered Double Hydroxides (LDHs) hold promise regarding their successful utilization in practical applications due to their interesting electrochemical performance. This study aims to elucidate the effect of LDH introduction on the internal arrangement of water molecules in the hydrophilic clusters of sPSU and on its proton transport properties. Swelling tests, NMR characterization, and Electrochemical Impedance Spectroscopy (EIS) investigation allowed us to demonstrate that LDH platelets act as physical crosslinkers between -SO3H groups of adjacent polymer chains. This increases dimensional stability while simultaneously creating continuous paths for proton conduction. This feature, combined with its impressive water retention capability, allows sPSU to yield a proton conductivity of ca. 4 mS cm-1 at 90 °C and 20% RH.
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In this article, the specific features of competitive ionic and molecular transport in nanocomposite systems based on network membranes synthesized by radical polymerization of polyethylene glycol diacrylate in the presence of LiBF4, 1-ethyl-3-methylimidazolium tetrafluoroborate, ethylene carbonate (EC), and TiO2 nanopowder (d~21 nm) were studied for 1H, 7Li, 11B, 13C, and 19F nuclei using NMR. The membranes obtained were studied through electrochemical impedance, IR-Fourier spectroscopy, DSC, and TGA. The ionic conductivity of the membranes was up to 4.8 m Scm-1 at room temperature. The operating temperature range was from -40 to 100 °C. Two types of molecular and ionic transport (fast and slow) have been detected by pulsed field gradient NMR. From quantum chemical modeling, it follows that the difficulty of lithium transport is due to the strong chemisorption of BF4- anions with counterions on the surface of TiO2 nanoparticles. The theoretical conclusion about the need to increase the proportion of EC in order to reduce the influence of this effect was confirmed by an experimental study of a system with 4 moles of EC. It has been shown that this approach leads to an increase in lithium conductivity in an ionic liquid medium, which is important for the development of thermostable nanocomposite electrolytes for Li//LiFePO4 batteries with a base of lithium salts and aprotonic imidasolium ionic liquid.
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Charge transport, diffusion properties, and glassy dynamics of blends of imidazolium-based ionic liquid (IL) and the corresponding polymer (polyIL) were examined by Pulsed-Field-Gradient Nuclear Magnetic Resonance (PFG-NMR) and rheology coupled with broadband dielectric spectroscopy (rheo-BDS). We found that the mechanical storage modulus (G') increases with an increasing amount of polyIL and G' is a factor of 10,000 higher for the polyIL compared to the monomer (GIL'= 7.5 Pa at 100 rad s-1 and 298 K). Furthermore, the ionic conductivity (σ0) of the IL is a factor 1000 higher than its value for the polymerized monomer with 3.4×10-4 S cm-1 at 298 K. Additionally, we found the Haven Ratio (HR) obtained through PFG-NMR and BDS measurements to be constant around a value of 1.4 for the IL and blends with 30 wt% and 70 wt% polyIL. These results show that blending of the components does not have a strong impact on the charge transport compared to the charge transport in the pure IL at room temperature, but blending results in substantial modifications of the mechanical properties. Furthermore, it is highlighted that the increase in σ0 might be attributed to the addition of a more mobile phase, which also possibly reduces ion-ion correlations in the polyIL.
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Conventional Nafion membranes demonstrate a strong affinity for methanol, resulting in a high fuel crossover, poor mechanical stability, and thus poor performance in direct methanol fuel cells (DMFCs). This study involves the synthesis and physiochemical characterization of an alternative polymer electrolyte membrane for DMFCs based on sulfonated poly(ether ether ketone) and a layered double hydroxide (LDH) material. Nanocomposite membranes (sPL), with filler loading ranging between 1 wt% and 5 wt%, were prepared by simple solution intercalation and characterized by XRD, DMA, swelling tests, and EIS. For the first time, water and methanol mobility inside the hydrophilic channels of sPEEK-LDH membranes were characterized by NMR techniques. The introduction of LDH nanoplatelets improved the dimensional stability while having a detrimental effect on methanol mobility, with its self-diffusion coefficient almost two orders of magnitude lower than that of water. It is worth noting that anionic lamellae are directly involved in the proton transport mechanism, thus enabling the formation of highly interconnected paths for proton conduction. In this regard, sPL3 yielded a proton conductivity of 110 mS cm-1 at 120 °C and 90% RH, almost attaining the performance of the Nafion benchmark. The nanocomposite membrane also showed an excellent oxidative stability (over more than 24 h) during Fenton's test at 80 °C. These preliminary results demonstrate that an sPL3 nanocomposite can be potentially and successfully applied in DMFCs.
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Sulfonated Polysulfone (sPSU) is emerging as a concrete alternative to Nafion ionomer for the development of proton exchange electrolytic membranes for low cost, environmentally friendly and high-performance PEM fuel cells. This ionomer has recently gained great consideration since it can effectively combine large availability on the market, excellent film-forming ability and remarkable thermo-mechanical resistance with interesting proton conductive properties. Despite the great potential, however, the morphological architecture of hydrated sPSU is still unknown. In this study, computational and experimental advanced tools are combined to preliminary describe the relationship between the microstructure of highly sulfonated sPSU (DS = 80%) and its physico-chemical, mechanical and electrochemical features. Computer simulations allowed for describing the architecture and to estimate the structural parameters of the sPSU membrane. Molecular dynamics revealed an interconnected lamellar-like structure for hydrated sPSU, with ionic clusters of about 14-18 Å in diameter corresponding to the hydrophilic sulfonic-acid-containing phase. Water dynamics were investigated by 1H Pulsed Field Gradient (PFG) NMR spectroscopy in a wide temperature range (20-120 °C) and the self-diffusion coefficients data were analyzed by a "two-sites" model. It allows to estimate the hydration number in excellent agreement with the theoretical simulation (e.g., about 8 mol H2O/mol SO3- @ 80 °C). The PEM performance was assessed in terms of dimensional, thermo-mechanical and electrochemical properties by swelling tests, DMA and EIS, respectively. The peculiar microstructure of sPSU provides a wider thermo-mechanical stability in comparison to Nafion, but lower dimensional and conductive features. Nonetheless, the single H2/O2 fuel cell assembled with sPSU exhibited better features than any earlier published hydrocarbon ionomers, thus opening interesting perspectives toward the design and preparation of high-performing sPSU-based PEMs.