Spatially resolved ultra-trace analysis of elements combining resonance ionization with a MALDI-TOF spectrometer

A combined setup for spatially resolved mass analysis of trace amounts of elements and macromolecules is presented. Using a MALDI-TOF mass spectrometer, a laser spectroscopic setup for resonant ionization of neutral atoms has been implemented. This allows for an efficient and selective detection of trace elements by means of resonance ionization mass spectrometry (RIMS). The instrumental scheme is described, and methodological developments are presented. In a first application pure, laser desorption/ionization with TOF-MS was used to measure mass distributions of cosmic nanodiamonds. For further applications regarding the spatially resolved ultra-trace analysis of elements in solid samples, an implanted target was used to characterize both laser desorption/ionization and laser desorption/resonance ionization for the detection of trace elements within. A perspective of the setup is given and future investigations are outlined.     

A New Method to Examine Interfacial Reactions of a Multilayered System NiAl–Hf–hBN on a Sapphire Fibre

Fibre reinforced NiAl offers new possibilities for the development of high strength structural materials of low density applicable in gas turbines at high operating temperatures. The properties of composite materials are strongly influenced by the strength of the fibre–matrix interface. In addition, if fibre and matrix differ in their thermal expansion coefficients, a well controlled interface reaction at high temperature changes is demanded. Therefore, two layers consisting of BN and Hf were embedded between a sapphire fibre and NiAl and heated at 1350 °C to find a compromise between adhesion and ductility. The control and characterization of the reaction zone is essential for the development of these new materials. Especially, the characterization of the fibre-coating interface is a challenge. The different hardness of fibre and coating makes it nearly impossible to use a conventional cross-section preparation. Further, the small dimension of the reaction zone requires the use of analytical techniques providing high lateral resolution. In order to accomplish these requirements, a newly developed technique FIB (Focused Ion Beam)-EPMA (Electron Probe Microanalysis) was combined with XRD (X-ray diffraction). XRD was performed for the identification of the phases. The reaction zone was exposed by a special FIB preparation technique and examined by surface-sensitive EPMA. This allowed to determine the spatial distribution of the different phases.     

Straightforward Approach for Preparing Durable Antibacterial ZnO Nanoparticle Coatings on Flexible Substrates

Flexible antibacterial materials have gained utmost importance in protection from the distribution of bacteria and viruses due to the exceptional variety of applications. Herein, we demonstrate a readily scalable and rapid single-step approach for producing durable ZnO nanoparticle antibacterial coating on flexible polymer substrates at room temperature. Substrates used are polystyrene, poly(ethylene-co-vinyl acetate) copolymer, poly(methyl methacrylate), polypropylene, high density polyethylene and a commercial acrylate type adhesive tape. The deposition was achieved by a spin-coating process using a slurry of ZnO nanoparticles in toluene. A stable modification layer was obtained when toluene was a solvent for the polymer substrates, namely polystyrene and poly(ethylene-co-vinyl acetate). These coatings show high antibacterial efficiency causing >5 log decrease in the viable counts of Gram-negative bacteria Escherichia. coli and Gram-positive bacteria Staphylococcus aureus in 120 min. Even after tapping these coated surfaces 500 times, the antibacterial properties remained unchanged, showing that the coating obtained by the presented method is very robust. In contrast to the above findings, the coatings are unstable when toluene is not a solvent for the substrate.     

Reorganization free energies for long-range electron transfer in a porphyrin-binding four-helix bundle protein

To explore the possibility of electron transport in a recently designed four-helix bundle protein (Cochran, F. V.; et al. J. Am. Chem. Soc. 2005, 127, 1346), we have computed the reorganization free energy for (i) oxidation of a single Ru-porphyrin cofactor and (ii) electron self-exchange between two Ru-porphyrin cofactors binding to the solvated protein. Sampling the classical electrostatic energy gap for 20 ns, we find that the fluctuations are well described by Gaussian statistics and obtain reorganization free energies of 0.90 +/- 0.04 eV for oxidation and 1.36 +/- 0.08 eV for self-exchange. The latter is 0.1-0.2 eV higher than the experimental estimate for interprotein electron self-exchange in cytochrome b5. As in natural electron carriers, inner-sphere reorganization is very small, 88 meV for self-exchange between two model cofactors computed at the density functional level of theory. Decomposing the outer-sphere reorganization free energy, we find that the solvent (aqueous ionic solution) is the primary outer-sphere medium for oxidation, contributing 0.60 eV (69%). The protein contributes only 0.27 eV (31%). For self-exchange, the solvent contribution, 0.68 eV (54%), and the protein contribution, 0.59 eV (46%), are almost equally important. The large solvent contribution is due to the slow decay of dipole reorientation of the solvent as a function of distance to the cofactor, implying that the change in the electric field upon electron transfer is not as effectively screened by the four-helix bundle protein. However, ranking the residues according to their free energy contributions, it is suggested that the reorganization free energy can be decreased by about 0.2 eV if two glutamine residues in the vicinity of the cofactor are mutated into less polar amino acids.     

Boride-based nano-laminates with MAX-phase-like behaviour

MAX-phases being usually composed of transition metals, group A elements and carbon/nitrogen are considered interesting materials for many applications because of their tremendous bulk modulus, “reversible” plasticity, and machinability. This is mainly due to their unique kind of bonding comprising covalent, ionic as well as metallic bonds providing “easy” planes of rupture and deformability due to the layered crystal structures.In transition metal boride systems, similar types of bonding are available. In particular the W₂B₅-structure type and its stacking variations allow the synthesis of strongly layered crystal structures exhibiting unique delamination phenomena.The paper presents ab initio calculations showing the similarities of bonding between the ternary carbides and the corresponding ternary or quaternary borides. Formation of boride-based nano-laminates from auxiliary liquid phases, from the melt as well as during sintering and precipitation from supersaturated solid solutions will be discussed by means of SEM and TEM studies. The role of impurities weakening the interlayer bonding will be addressed in particular. The pronounced cleavage parallel to the basal plane gives rise for crack deflection and pull-out mechanisms if the laminates are dispersed in brittle matrices such as boron carbide, silicon carbide or other transition metal borides.     

Chemie polyfunktioneller liganden LXIII. Adamantankäfigverbindungen mit den heteroelementen arsen,stickstoff und silizium

The reaction of 1,1,1-tris(diiodarsinomethyl)ethane, CH₃C(CH₂AsI₂)₃ (I), with i-C₃H₇NH₂, n-C₄H₉NH₂, C₆H₅NH₂, p-CH₃C₆H₄NH₂ and [(CH₃)₃Si]₂NH in the presence of (C₂H₅)₃N as auxiliary base in THF gives the adamantane cage compounds CH₃C(CH₂AsNC₃H₇)₃ (III), CH₃C(CH₂AsNC₄H₉)₃ (IV), CH₃C(CH₂AsNC₆H₅)₃ (V), CH₃C(CH₂AsNC₆H₄CH₃)₃ (VI) and CH₃C[CH₂AsNSi(CH₃)₃]₃ (VII). VII is also obtained in the reaction of I with NaN[Si(CH₃)₃]₂. The by-product (CH₃)₃SiO(CH₂)₄I (VIII) could be isolated in both syntheses of VII. All compounds have been characterized by mass spectrometry and infrared, Raman and ¹H NMR spectroscopy.     

NMR spectroscopic characterization of millisecond protein folding by transverse relaxation dispersion measurements

The cold shock protein CspB adopts its native and functional tertiary structure on the millisecond time scale. We employed transverse relaxation NMR methods, which allow a quantitative measurement of the cooperativity of this fast folding reaction on a residue basis. Thereby, chemical exchange contributions to the transverse relaxation rate (R(2)) were observed for every residue of CspB verifying the potential of this method to identify not only local dynamics but also to characterize global events. Toward this end, the homogeneity of the transition state of folding was probed by comparing Chevron plots (i.e., dependence of the apparent folding rate on the denaturant concentration) determined by stopped-flow fluorescence with Chevron plots of six residues acquired by R(2) dispersion experiments. The coinciding results obtained for probes at different locations in the three-dimensional structure of CspB indicate the ability and significance of transverse relaxation NMR to determine Chevron plots on a residue-by-residue basis providing detailed insights on the nature of the transition state of folding.     

Generic method for systematic phase selection and method development of biochromatographic processes. Part I. Selection of a suitable cation-exchanger for the purification of a pharmaceutical protein

Even if the first protein therapeutics are now for more than 20 years on the market the selection of suitable adsorbents for the preparative downstream processing (DSP) of these biomolecules as well as the method development towards process conditions are still based mainly on 'trial and error'. Therefore, theses processes are not perfectly efficient, but indeed very time consuming and laborious. In this study a novel systematic method is introduced to find a suitable adsorbent (not necessarily the best one) with appropriate separation parameters for a specific separation with reduced effort. Following this strategy, the adsorbents must first be packed into columns under preparative conditions and then characterized completely with regard to, e.g. pressure drop, k'-values, plate heights (HETP curves), selectivity and capacity by using test substances, which are similar in their characteristics (molecular mass, size, charge distribution, hydrophobicity) to the target proteins. With the database once determined, a preselection of most suitable adsorbents including separation parameters is made regarding chromatographic and also economical properties. After this, preparative experiments must be conducted with a reduced number of adsorbents to figure out the individual influence of side components. This approach is demonstrated for the separation of an exemplary industrial protein mixture using cation-exchange chromatography (CEX). Characterization of different weak CEX-adsorbents is illustrated. After comparing these phases with each other, a first preselection and a prediction of suitable adsorbents is made. In the following preparative separation conditions (load, velocity, gradient) are determined for the preparative separations using the database and results of some additional experiments. The final comparison of separation performance in preparative scale confirms this selection and so the applicability of the new method.