Recent advances in the control of morphology and surface chemistry of porous polymer-based monolithic stationary phases and their application in CEC

This review focuses on developments in the field of polymer-based monolithic columns for CEC published in the literature since the beginning of the year 2005. The possibility of in-situ preparation as well as easy control over their porous properties and surface chemistries clearly make monolithic separation media an attractive alternative to capillary columns packed with particles. Different variables such as polymerization conditions, morphology, and surface chemistry are shown to directly affect performance of monolithic capillary columns in terms of efficiency, analysis time, and retention.     

Photopolymerized monolithic capillary columns for rapid micro high-performance liquid chromatographic separation of proteins

The preparation of monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns using photoinitiated in situ polymerization within 200 microm i.d. capillaries and their application for microHPLC separations of proteins have been studied. The low resistance to flow characteristic of monolithic columns, enabled the use of very high flow rates of up to 100 microL/min representing a flow velocity of 87 mm/s. Very good separations of a model protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin was achieved in less than 40 s using a very simple single step gradient of the mobile phase. Interestingly, no effect of the pore size on the separations of proteins was observed for these monolithic columns within the size range of 0.66-2.2 microm. The monolithic microHPLC columns are found very robust and no changes in the long term separation performance and back pressure were observed.     

An analysis of avidin, biotin and their interaction at attomole levels by voltammetric and chromatographic techniques

The electroanalytical determination of avidin in solution, in a carbon paste, and in a transgenic maize extract was performed in acidic medium at a carbon paste electrode (CPE). The oxidative voltammetric signal resulting from the presence of tyrosine and tryptophan in avidin was observed using square-wave voltammetry. The process could be used to determine avidin concentrations up to 3 fM (100 amol in 3 l drop) in solution, 700 fM (174 fmol in 250 l solution) in an avidin-modified electrode, and 174 nM in a maize seed extract. In the case of the avidin-modified CPE, several parameters were studied in order to optimize the measurements, such as electrode accumulation time, composition of the avidin-modified CPE, and the elution time of avidin. In addition, the avidin-modified electrode was used to detect biotin in solution (the detection limit was 7.6 pmol in a 6 l drop) and to detect biotin in a pharmaceutical drug after various solvent extraction procedures. Comparable studies for the detection of biotin were developed using HPLC with diode array detection (HPLC-DAD) and flow injection analysis with electrochemical detection, which allowed biotin to be detected at levels as low as 614 pM and 6.6 nM, respectively. The effects of applied potential, acetonitrile content, and flow rate of the mobile phase on the FIA-ED signal were also studied.     

Electron super-rich radicals in the gas phase. A neutralization-reionization mass spectrometric and ab initio/RRKM study of diaminohydroxymethyl and triaminomethyl radicals

Diaminohydroxymethyl (1) and triaminomethyl (2) radicals were generated by femtosecond collisional electron transfer to their corresponding cations (1+ and 2+, respectively) and characterized by neutralization-reionization mass spectrometry and ab initio/RRKM calculations at correlated levels of theory up to CCSD(T)/aug-cc-pVTZ. Ion 1+ was generated by gas-phase protonation of urea which was predicted to occur preferentially at the carbonyl oxygen with the 298 K proton affinity that was calculated as PA = 875 kJ mol-1. Upon formation, radical 1 gains vibrational excitation through Franck-Condon effects and rapidly dissociates by loss of a hydrogen atom, so that no survivor ions are observed after reionization. Two conformers of 1, syn-1 and anti-1, were found computationally as local energy minima that interconverted rapidly by inversion at one of the amine groups with a <7 kJ mol-1 barrier. The lowest energy dissociation of radical 1 was loss of the hydroxyl hydrogen atom from anti-1 with ETS = 65 kJ mol-1. The other dissociation pathways of 1 were a hydroxyl hydrogen migration to an amine group followed by dissociation to H2N-C=O* and NH3. Ion 2+ was generated by protonation of gas-phase guanidine with a PA = 985 kJ mol-1. Electron transfer to 2+ was accompanied by large Franck-Condon effects that caused complete dissociation of radical 2 by loss of an H atom on the experimental time scale of 4 mus. Radicals 1 and 2 were calculated to have extremely low ionization energies, 4.75 and 4.29 eV, respectively, which belong to the lowest among organic molecules and bracket the ionization energy of atomic potassium (4.34 eV). The stabilities of amino group containing methyl radicals, *CH2NH2, *CH(NH2)2, and 2, were calculated from isodesmic hydrogen atom exchange with methane. The pi-donating NH2 groups were found to increase the stability of the substituted methyl radicals, but the stabilities did not correlate with the radical ionization energies.     

Photo-induced ligand substitution at a remote site via electron transfer in a porphyrin-appended rhenium carbonyl supermolecule

The photochemical and electrochemical properties of a Zn-porphyrin appended rhenium(I) tricarbonyl bipyridine 3-Me-pyridine complex have been investigated; visible-light sensitisation of electron transfer results in ligand substitution at a site remote from the chromophore.     

A convenient route to diazepines by intramolecular cyclisation of carbonylazides

The appropriate substituted carbonylazides I treated with warm acetic acid give the corresponding acetamide IV or the cyclic lactam VI . This is a convenient route to 10,11-Dihydro-5H-pyrrolo[1,2-a][1,4]-benzodiazepin-11-one and 10,11-Dihydro-4H-thieno[2,3-e]pyrrolo[1,2-a][1,4]diazepin-10-one.     

Microscopy and single molecule detection in photosynthesis

Progress in various fields of microscopy techniques brought up enormous possibilities to study the photosynthesis down to the level of individual pigment-protein complexes. The aim of this review is to present recent developments in the photosynthesis research obtained using such highly advanced techniques. Three areas of microscopy techniques covering optical microscopy, electron microscopy and scanning probe microscopy are reviewed. Whereas the electron microscopy and scanning probe microscopy are used in photosynthesis mainly for structural studies of photosynthetic pigment-protein complexes, the optical microscopy is used also for functional studies.     

Response of a Si-diode-based device to fast neutrons

Semiconductor devices based on a Si-detector are frequently used for charged particle's detection; one application being in the investigation of cosmic radiation fields. From the spectra of energy deposition events in such devices, the total energy deposited by the radiation in silicon can be derived. This contribution presents the results of studies concerning the response of this type of detector to fast neutrons. First, the spectrum of energy deposition was established in fast neutron radiation fields with average energies from 0.5 to 50 MeV. It was found that these spectra vary significantly with the neutron energy. The comparison with the spectra registered in photon beams permitted an estimation of the part of energy deposited that could be attributed to neutrons. It was found that this part increases rapidly with neutron energy. The possibilities to use this type of detector for neutron detection and dosimetry for radiation protection are analysed and discussed.     

Monolithic columns with a gradient of functionalities prepared via photoinitiated grafting for separations using capillary electrochromatography

Stationary phases for capillary electrochromatography with a longitudinal gradient of functionalities have been prepared via photoinitiated grafting of polymer chains onto the pore surface of a porous polymer monolith. In order to achieve the desired retention and electroosmotic flow, the hydrophobic poly(butyl methacrylate-co-ethylene dimethacrylate) monolith with optimized porous properties was grafted with a layer of ionizable polymer, poly(2-acrylamido-2-methyl-1-propanesulfonic acid). A moving shutter and a neutral density filter were used to control the dose of UV light received at different locations along the monolith in order to create the longitudinal gradient of functionalities. Formation of the desired gradients was confirmed using electron probe microanalysis of different locations along the column. The preparation technique significantly affects performance in the CEC mode as demonstrated on the separations of a model mixture using columns both with homogeneous distribution of grafts and with a gradient of functionality. Columns grafted with the gradient of functionalities were found superior to those functionalized uniformly. A comparison of the performance of the gradient column with another containing evenly distributed functionalities showed the performance benefits of the "gradient" column.     

A Pencil Graphite Electrode In Situ Modified by Monovalent Copper: a Promising Tool for the Determination of Methylxanthines

A pencil graphite electrode (PeGE) exhibits a promising tool for the electrochemical analysis of xanthine (Xan) and its N‐methyl derivatives (1‐, 3‐, 7‐ and 9‐mXan). The changes in their level in blood, serum, urine, as products of purine catabolism, can indicate the development of some diseases. Sensitivity‐enhanced voltammetric detection of mXans was achieved by forming of complex with Cu(I) and application of elimination procedure. The Cu(I)‐mXan complex was identified by means of titration of electrochemically produced cuprous ions by mXan. Our approach enables separation of overlapped mXan oxidation signals. Based on the obtained results, we found that the effect of methyl group position on the xanthine skeleton was significant and it was also discussed.