Gas-phase models for catalysis: alkane activation and olefin epoxidation by the triatomic cation Ag2O+

Electrospray ionization of aqueous silver nitrate is used for the preparation of the disilver-oxide cation Ag2O+ in the gas phase. The mass-selected cation is capable of activating C-H bonds of simple alkanes other than methane via H-atom abstraction, i.e., Ag2O+ + R-H --> Ag2OH+ + R* (R = C2H5, C3H7, C4H9). Clean O-atom transfer from Ag2O+ is observed with ethene as a neutral reagent, whereas oxygenation and allylic C-H abstraction compete in the case of propene. The gaseous Ag2O+ cation can thus be regarded as a minimalist model for the problems associated with the silver-mediated epoxidation of olefins more complex than ethene itself. The experimental findings are fully supported by the results of quantum chemical studies, thereby providing deep mechanistic insight into the reactions in the idealized gas phase, which also might have implications for further improvements in applied catalysis.     

Bond-formation versus electron transfer: C-C-coupling reactions of hydrocarbon dications with benzene

The bimolecular reactions of several hydrocarbon dications C(m)H(n)(2+) (m = 6-10, n = 4-9) with neutral benzene are investigated by tandem mass spectrometry using a multipole instrument. Not surprisingly, the major reaction of C(m)H(n)(2+) with benzene corresponds to electron transfer from the neutral arene to the dication resulting in the pair of monocationic products C(m)H(n)(+) + C(6)H(6)(+). In addition, also dissociative electron transfer takes place, whereas proton transfer from the C(m)H(n)(2+) dication to neutral benzene is almost negligible. Interestingly, the excess energy liberated upon electron transfer from the neutral arene to the C(m)H(n)(2+) dication is not equally partitioned in the monocationic products in that the cations arising from the dicationic precursor have a higher internal energy content than the monocations formed from the neutral reaction partner. In addition to the reactions leading to monocationic product ions, bond-forming reactions with maintenance of the two-fold charge are observed, which lead to a condensation of the C(m)H(n)(2+) dications with neutral benzene under formation of intermediate C(m+6)H(n+6)(2+) species and then undergo subsequent losses of molecular hydrogen or neutral acetylene. This reaction complements a recently proposed dicationic route for the formation of polycyclic aromatic hydrocarbons under extreme conditions such as they exist in interstellar environments.     

Bimolecular reactions of molecular dications: reactivity paradigms and bond-forming processes

The bimolecular reactivity of molecular dications in the gas phase is reviewed from an experimental point of view. Recent research has demonstrated that in addition to the ubiquitous occurrence of electron transfer in the reactions of gaseous dications with neutral molecules, bond-forming reactions play a much larger role than anticipated before. Thus, quite a number of hydrogen-containing dications show proton transfer to neutral reagents as an abundant or even as the major pathway, and also the nature of the neutral reagent itself is decisive for the amount of proton transfer which takes place. Further, several hydrocarbon dications C(m)H(n)(2+) of medium size (m = 6-14, n = 6-10) undergo bond-forming reactions with unsaturated hydrocarbons such as acetylene or benzene, thereby offering new routes for the formation of larger aromatic compounds under extreme conditions such as interstellar environments. Likewise, recent results on the bimolecular reactivity of multiply charged metal ions have revealed the occurrence of a number of new bond-forming reactions which open promising prospects for further research.     

A novel microbial biosensor based on cells of <Emphasis Type="Italic">Gluconobacter oxydans</Emphasis> for the selective determination of 1,3-propanediol in the presence of glycerol and its application to bioprocess monitoring

Novel and selective microbial amperometric biosensors that use Gluconobacter oxydans cells to monitor the bacterial bioconversion of glycerol (Gly) to 1,3-propanediol (1,3-PD) are described. Two different mediators, ferricyanide and flexible polyvinylimidazole osmium functionalized polymer (Os-polymer), were employed to prepare two different microbial biosensors, both of which gave high detection performance. The good operational stabilities of both types of biosensor were underlined by the ability to detect 1,3-PD throughout 140 h of continuous operation. Both microbial biosensor systems showed excellent selectivity for 1,3-PD in the presence of a high excess of glycerol [selectivity ratios (1,3-PD/Gly) of 118 or 245 for the ferricyanide and Os-polymer systems, respectively]. Further, the robustness of each microbial biosensor was highlighted by the high reliability of 1,3-PD detection achieved (average RSD of standards <2%, and well below 4% for samples). The biosensor implementing the Os-polymer mediator exhibited high selectivity towards 1,3-PD detection and allowed moderate sample throughput (up to 12 h⁻¹) when integrated into a flow system. This system was used to monitor the concentration of 1,3-PD during a real bioprocess. Results from biosensor assays of 1,3-PD in bioprocess samples taken throughout the fermentation were in a very good agreement with results obtained from reference HPLC assays (R ² = 0.999).     

Controlled photostability of luminescent nanocrystalline ZnO solution for selective detection of aldehydes

Water-soluble, silane-functionalized ZnO nanocrystals were synthesized with improved colloidal stability, and their photostability was controlled for the selective detection of aldehydes.     

Low-frequency magnetic field effect on cytoskeleton and chromatin

The effect of magnetic fields on the living systems is studied in vivo or in vitro in very broad spectrum of organisms, cells and tissues. The mechanism of their acting is not known until now. We studied low-frequency magnetic field effect on cytoskeleton and on the structure of chromatin in human cells. We used cell line of small lung carcinoma (A549) and the effects of magnetic field on cytoskeleton and higher-order chromatin structure were analyzed 96 h of magnetic field exposure. Magnetic field generated by the cylindrical soil was homogenous and the cells were cultivated at 37 degrees C in humidified atmosphere containing 5% CO(2). Magnetic field induction was B(m)=2 mT and the net frequency f=50 Hz. In such affected and control cells the F-actin was estimated using FITC-conjugated Phalloidin and mitochondria were studied using MitoTracker (Molecular Probes). Images of cytoskeleton and genetic loci were acquired using confocal microscopy and analysis was performed by FISH 2.0 software. Slight morphological changes of F-actin filaments and mitochondria were observed in affected cells and nuclear condensation was found. These effects could be related to the process of cell death apoptosis probably induced by magnetic field. The studies aimed at centromeric heterochromatin (9cen) did not show statistically significant changes. Therefore, we suggest that magnetic field has no influence on higher order chromatin structure but certain changes could be observed on the level of cytoskeleton. However, these statements need a thorough verification. Our preliminary experiments will be extended and the effect of magnetic field on another structures of cytoskeleton and cell nuclei will be further studied.     

Novel approaches to the analysis of steroid estrogens in river sediments

A wide range of estrogenic contaminants has been detected in the aquatic environment. Among these, natural and synthetic steroid estrogens, typically present in municipal sewage-treatment plant (STP) effluents, are the most potent. In this study a new GC–MS method has been developed for direct analysis of five major steroid estrogens (estrone, 17β-estradiol, 17α-ethinylestradiol, dienestrol, and diethylstilbestrol) in river sediments. Four GC–MS systems used for analysis of underivatized analytes in purified extracts were compared. Relatively low detection limits (1.5–5 ng g⁻¹ dried sediment) and good repeatability of GC splitless injection (RSD 1–2%) were achieved by use of a system combining low-pressure gas chromatography with a single-quadrupole mass analyzer (LP-GC–MS). Use of orthogonal gas chromatography (GC×GC) hyphenated with high-speed time-of-flight mass spectrometry (HSTOF-MS) enabled not only significantly better resolution of target analytes, and their unequivocal identification, but also further improvement (decrease) of their detection limits. In addition to these outcomes, use of this unique GC×GC–TOF-MS system enabled identification of several other non-target chemicals, including pharmaceutical steroids, present in purified sediment extracts.     

On-line preconcentration of weak electrolytes by electrokinetic accumulation in CE: experiment and simulation

A new on-line preconcentration technique was developed that makes possible to determine nanomolar concentrations of weak acidic analytes in CE. The method consists of long-running electrokinetic sample injection and stacking (electrokinetic immobilization) of the analytes at a boundary of two electrolytes with different pH values (pH 9.5 and 2.5) and consequent mobilization of the stacked uncharged analytes in a micellar system (containing SDS micelles). Several factors including buffer concentration, pH, applied voltage, time of preconcentration, and SDS concentration were tested to optimize the analysis method. An about 4600-fold increase of the sample concentration (in comparison with the standard CZE) can be achieved during the preconcentration step. Two preservatives applied in food industry -- benzoic acid and sorbic acid were used as model samples. The applicability of the proposed method in food analysis was demonstrated by determination of nanomolar concentrations of benzoic acid in sunflower oil. An extended version of the computer program Simul was used for modeling both the preconcentration and mobilization processes taking place in the capillary.     

Assay of tyrosine hydroxylase based on high-performance liquid chromatography separation and quantification of L-dopa and L-tyrosine

An assay of L-tyrosine (Tyr) hydroxylating activity operating in lincomycin biosynthesis is described. The assay development consisted of HPLC procedure development, assessing the effect of reaction mixture components on non-enzymatic Dopa and Tyr oxidation, and sample stability evaluation. The HPLC procedure with isocratic elution and fluorescence detection was developed and validated. The method showed a wide linear range of Dopa determination of 0.125-25 micromol/L with lower limit of quantification (LLOQ) of 0.125 micromol/L, RSD of 7.2% and accuracy of 101.7%. The studied linear range of Tyr was 15.625 mmol/L to 500 mmol/L with LLOQ of 15.625 mmol/L, RSD of 1.1%, and accuracy of 98.1%. Recoveries for Dopa and Tyr were 100.66 +/- 0.89% and 94.76 +/- 0.94%, respectively. The inter- and intra-day accuracies and precisions were all within 10%. Samples of the reaction mixture were stable for at least 24 h at room temperature (RT) and 28 days at -20 degrees C. The method was tested for the enzyme activity monitoring in purified as well as crude preparations and enabled micro preparation of the enzyme product during confirmation of its identity. The influence of pH and ascorbic acid content in reaction mixture was studied with respect to non-enzymatic Tyr oxidation.     

Chiral superbases: the proton affinities of 1- and 2-aza[6]helicene in the gas phase

The proton affinities (PAs) of 1- and 2-azahelicene were determined using various mass spectrometric techniques and complementary results from density functional theory. With PAs of about 1000 kJ mol(-1), the helical backbone of both compounds offer promising perspectives for future research on enantioselective reactions of these helical nitrogen bases.