Synthesis,characterization and crystal structure analysis of a novel oxo-centered mixed-metal complex containing unsaturated bridging carboxylates

A novel oxo-centered trinuclear mixed-metal carboxylate complex with unsaturated bridging ligands [Fe₂Cr(μ₃-O)(C₃H₃O₂)₆(H₂O)₃]·NO₃·4H₂O has been synthesized and characterized by means of Elemental analyses, Infrared spectroscopy and Crystal structure analysis. The compound crystallizes isotypically in the monoclinic space group type P2₁/c. In the compound, each M^(III) cation is coordinated by six O atoms from four unsaturated carboxylate groups as bridging ligands, one water molecule as the terminal ligand, and a μ₃-oxygen atom in the center of an equilateral triangle. The infrared spectra show resolved bands arising from ν_asym(COO) and ν_sym(COO) vibration of bridging carboxylate ligands along with those of ν_asym(M₂M′O) vibration in the complex. The difference between symmetrical and asymmetrical (COO) ligands indicate that the acrylate bridge is present in the structure of complex.      

Systematic studies on the determination of Hg-labelled proteins using laser ablation-ICPMS and isotope dilution analysis

A method was developed for the precise and accurate determination of ovalbumin labelled with p-hydroxy-mercuribenzoic acid (pHMB) using polyacrylamide gel electrophoresis with ns-laser ablation–inductively coupled plasma mass spectrometry. Following systematic optimisation of the ablation process in terms of detection sensitivity, two different quantification strategies were applied: external calibration using standards of the derivatized protein after ¹³C⁺ normalization and, as a proof of concept, label-specific isotope dilution analysis (IDA) using pHMB enriched in the isotope ¹⁹⁹Hg. Due to the inhomogeneous distribution of the protein within the gel bands, it could be demonstrated that the IDA approach was superior in terms of precision and accuracy. Furthermore, it permits a reliable quantification, if more complex separation protocols are applied, as typically occurring analyte loss and degradation can be compensated for as soon as complete mixture of spike and sample is achieved. The estimated limit of detection was 160 fmol in the case of ovalbumin. In contrast to earlier studies using metals naturally present in proteins, no loss of mercury was observed during separation under denaturing conditions and other sample preparation steps. Using label-specific IDA, the measured isotope ratios in the gel corresponded to recoveries between 95% and 103%.     

Fabrication of carbon nanofiber by pyrolysis of freeze-dried cellulose nanofiber

The possibility of fabricating carbon nanofibers from cellulose nanofibers was investigated. Cellulose nanofiber of ~50 nm in diameter was produced using ball milling in an eco-friendly manner. The effect of the drying techniques of cellulose nanofibers on the morphology of carbon residue was studied. After pyrolysis of freeze-dried cellulose nanofibers below 600 °C, amorphous carbon fibers of ~20 nm in diameter were obtained. The pyrolysis of oven-dried precursors resulted in the loss of original fibrous structures. The different results arising from the two drying techniques are attributed to the difference in the spatial distance between cellulose nanofiber precursors.     

Novel bound states treatment of the two dimensional Schrödinger equation with pseudocentral potentials plus multiparameter noncentral potential

By converting the rectangular basis potential V(x, y) into the form as \({V({r}) + V({r},\varphi)}\) described by the pseudo central plus noncentral potential, particular solutions of the two dimensional Schrödinger equation in plane-polar coordinates have been carried out through the analytic approaching technique of the Nikiforov and Uvarov. Both the exact bound state energy spectra and the corresponding bound state wavefunctions of the complete system are determined explicitly and in closed forms. Our presented results are identical to those of the previous works and they may also be useful for investigation and analysis of structural characteristics in a variety of quantum systems.     

Development and validation of a rapid LC-ESI-MS/MS method for quantification of fluoxetine and its application to MS binding assays

In the present study, a rapid and sensitive LC-ESI-MS/MS method for quantification of (S)-fluoxetine as a native marker in mass spectrometry (MS) binding assays addressing the human serotonin transporter (hSERT) was developed and validated. The concept of MS binding assays based on mass spectrometric quantification of a nonlabeled marker recently introduced by us represents a promising alternative to conventional radioligand binding without the drawbacks inherently connected with radioisotope labeling. For high-performance liquid chromatography (HPLC), a 20 × 2-mm RP-18 column with a mobile phase composed of acetonitrile and ammonium bicarbonate buffer (5 mmol L⁻¹, pH 9.5) at a ratio of 80:20 (v/v) and a flow rate of 800 μL min⁻¹ in an isocratic mode were used, resulting in a chromatographic cycle time of 60 s. Employing [²H₅]fluoxetine as internal standard enabled ESI-MS/MS quantification of (S)-fluoxetine between 3 nmol L⁻¹ and 50 pmol L⁻¹ (LLOQ) in matrix obtained from binding experiments without the need of any sample preparation. Validation of the method showed that linearity, intra-, and inter-batch accuracy as well as precision meet the requirements of the FDA guidance for bioanalytical method validation. Considering sensitivity and speed, the established method is clearly superior to those published for biological matrices so far. Furthermore, the method was transferred to other RP-18 columns of different lengths and respective validation experiments demonstrated its versatility and chromatographic robustness. Finally, the newly developed method was successfully applied to MS binding assays for hSERT. The affinity determined for (S)-fluoxetine in saturation experiments was in good agreement with literature data obtained in respective radioligand binding assays.     

Selective detection of dopamine with poly(diphenylamine sulfonic acid) modified electrode in the presence of ascorbic acid

A glassy carbon electrode was modified with electropolymerized film of diphenylamine sulfonic acid (DPASA). Electropolymerization was performed by cyclic voltammetry in 0.1 M KCl solution. The modified electrode showed an excellent electrocatalytic effect towards oxidation of dopamine (DA) and ascorbic acid (AA). Electrostatic interaction between the negatively charged poly(DPASA) film and either cationic DA species or anionic AA species favorably contributed to the redox response of DA and AA. Anodic peaks of DA and AA in their mixture were well separated by ca 168 and −11.8 mV. The proposed modified electrode was utilized for selective determination of dopamine in the concentration range of 5.0 × 10^t7–2.0 × 10⁻⁵ M in the presence of high concentration of ascorbic acid. Detection limit was 6.5 × 10⁻⁹ M.     

A liquid drop RC filter apparatus for detection

A new analytical detector based on a liquid drop resistor–capacitor (RC) filter is described, in which transformed gain vs. frequency curves are used to analyze compounds. This detector can be used to detect either charged or neutral species (that are dielectrically different) which are dissolved in a liquid (e.g., water, alcohol, solvent mixtures, etc.). This device was fabricated by modifying an electrowetting on dielectric (EWOD)-based experimental setup. When a liquid drop is placed on a dielectric surface, the system acts as a RC filter. At a given frequency, gain is a function of conductivity, surface tension, dielectric constant, double-layer thickness of the solid–liquid drop interface, as well as the applied voltage. Since different liquids and solutions have different physical properties, each liquid/solution has a unique curve (peak) in gain vs. frequency plot. This is the basic principle behind the detector. Different amounts of zinc chloride dissolved in water, benzalkonium chloride in water, 1-methylimidazole in water, cetyltrimethyl-ammonium chloride (CTAC) in water, and CTAC dissolved in ethylene glycol solutions were tested with the detector as proof of principle. The device can be used as a stand-alone detector or can easily be coupled with droplet based microfluidic lab-on-a-chip systems such as EWOD-based microfluidic chips.     

Intriguing Differences in the Gas-Phase Dissociation Behavior of Protonated and Deprotonated Gonyautoxin Epimers

The aim of this study was to investigate the unusual gas-phase dissociation behavior of two epimer pairs of protonated gonyautoxins (GTX) following electrospray ionization in comparison to their deprotonated counterparts. The chemical structures of the investigated GTX1-4 variants vary in their substitution pattern at N-1 and the stereochemical orientation of the hydroxysulfate group at C-11 (11α for GTX1/2 versus 11β for GTX3/4). The direct comparison of mass spectra in positive and negative ion modes illustrated two distinct features: first, an intriguing difference between protonated 11α and 11β species, where 11α conformations exhibited almost complete dissociation of [M + H]⁺ ions via facile SO₃ elimination, while 11β species remained mostly intact as [M + H]⁺; and second, the lack of such differences for the deprotonated counterparts. In this study, we propose an acid-catalyzed elimination mechanism from density functional theory calculations, initiated by a proton transfer of a guanidinium proton to the hydroxysulfate group with simultaneous SO₃ release, which is only possible for the 11α conformation based on intramolecular distances. The same mechanism explains the lack of a comparable SO₃ loss in the negative ion mode. CID experiments supported this proposed mechanism for GTX1 and GTX2. Computational modeling of product ions seen in the CID spectra of GTX3 and GTX4 established that the lowest energy dissociation pathway for the 11β epimers is elimination of water with the possibility for further SO₃ release from the intermediate product. Experimental data for structurally analogous decarbamoyl gonyautoxins confirmed the evidence for the GTX compounds as well as the proposed elimination mechanisms.     

Reply to the comment by F. Malatesta on: “Factorizing of a concentration function for the mean activity coefficients of aqueous strong electrolytes into individual functions for ionic species” by A. Ferse and H.-O. Müller

The arguments of Malatesta (J Solution Chem 29:771–779, 2000; Fluid Phase Equil 295:244–248, 2010) exclude the experimental determination of individual ion activity coefficients. I agree that a measurement of single-ion activity coefficients is impossible. But the comment of Malatesta (J Solid State Electrochem (in press), 2011) in the connection with the purely mathematical procedure developed by Ferse and Müller (J Solid State Electrochem (in press), 2011) is senseless because there is no new aspect which is not also given in the paper of Ferse and Müller (J Solid State Electrochem (in press), 2011). All of the mentioned problems are already discussed and clarified in the publication by Ferse and Müller (J Solid State Electrochem (in press), 2011). The purely mathematical method is a possibility to obtain the concentration functions for the individual activity coefficients of the complementary ion species by factorizing a product function of the experimentally accessible concentration dependence of the mean activity coefficients to the required power.     

Mapping a Noncovalent Protein–Peptide Interface by Top-Down FTICR Mass Spectrometry Using Electron Capture Dissociation

Noncovalent protein–ligand and protein–protein complexes are readily detected using electrospray ionization mass spectrometry (ESI MS). Furthermore, recent reports have demonstrated that careful use of electron capture dissociation (ECD) fragmentation allows covalent backbone bonds of protein complexes to be dissociated without disruption of noncovalent protein–ligand interactions. In this way the site of protein–ligand interfaces can be identified. To date, protein–ligand complexes, which have proven tractable to this technique, have been mediated by ionic electrostatic interactions, i.e., ion pair interactions or salt bridging. Here we extend this methodology by applying ECD to study a protein–peptide complex that contains no electrostatics interactions. We analyzed the complex between the 21 kDa p53-inhibitor protein anterior gradient-2 and its hexapeptide binding ligand (PTTIYY). ECD fragmentation of the 1:1 complex occurs with retention of protein–peptide binding and analysis of the resulting fragments allows the binding interface to be localized to a C-terminal region between residues 109 and 175. These finding are supported by a solution-phase competition assay, which implicates the region between residues 108 and 122 within AGR2 as the PTTIYY binding interface. Our study expands previous findings by demonstrating that top-down ECD mass spectrometry can be used to determine directly the sites of peptide–protein interfaces. This highlights the growing potential of using ECD and related top-down fragmentation techniques for interrogation of protein–protein interfaces.