Separation of DABS derivatives of amino acids by multiple gradient development (MGD) in thin-layer chromatography

Summary A mixture of 13 DABS-amino acids has been chromatographed on high-performance silica gel layers developed with eluents containing increasing concentrations of ethyl acetate in heptane + chloroform, using a modification of stepwise multiple development MGD described in a earlier paper. Densitograms were obtained at 485 nm. The MGD method was very efficient, separating all 13 DABS-amino acids, and rapid, owing to the use of a non-aqueous mobile phase.     

Optimized Method for the Determination of Flavonoid Glycosides and Aglycones

The relationship R_M=f (log % S; S- modifier) of several flavonoids and their sugar derivatives was examined by using chromatographic systems chosen by preliminary experiments. The dependence for flavonoid aglycones was flat in the whole mobile phase concentration. Two types of the R_M=f (log % S) plots, related to the size of sugar molecule, were found. For glycosides with smaller sugar moiety (arabinose or glucose) the plot was almost parallel, while plots of derivatives with larger sugar moiety (rutose) were convergent. After plotting the relationship R_M=f (log % S), even if standards are not available, it is possible to qualify the examined compound as containing single or complex sugar moiety. For all mentioned above compounds the R_M=f (log %S) values were given. Silica gel Si60 was used as polar adsorbent. Mobile phase composition was determined in preliminary experiments; it was composed of two solvents: methanol in ethyl acetate in increasing concentration of methanol: 5 – 20%.     

Investigations on the current oscillations during the electrolytic reduction of dichromates in acetic solutions

The reduction mechanism at a mercury electrode of Eriochrome Cyanine R, in a 0.9 M NaClO₄+0.1 M HClO₄ supporting electrolyte, has been investigated by several electrochemical techniques. By means of coulometry at constant potential and cyclic voltammetry it was demonstrated that a radical is formed, which suffers disproportionation, after the first electron transfer. The results of the cyclic voltammetric investigation and the impedance measurements evidenced the intrinsic reversible nature of the electron transfers. Apparent irreversible polarographic behaviour is a consequence of the existence of chemical reactions following the electron transfers. The impedance measurements demonstrated the strong adsorption of the Ox and Red form of Eriochrome Cyanine R as well as of the radical formed. On the basis of the experimental data a reduction mechanism is proposed.     

Polarographic studies of catalytic reduction of hydrogen in Cr(VI)−As(III) and Cr(III)—As(III) systems

In the presence of traces of Cr(VI) or Cr(III) ions in ammonia or borate buffers containing the As(III) ions a catalytic hydrogen wave arises in the dc polarogram. It was established that the complex Cr(H₂AsO₃)_n^+3?n is formed in the solution, and that its reduced form adsorbed at DME is of catalytic activity. The wave can be employed for the determination of low concentrations (2×10^?8×10^?7M) of Cr(VI) and Cr(III) ions.     

Combining amperometry and mass spectrometry as a dual detection approach for capillary electrophoresis

Dual detection concepts (DDCs) are becoming more and more popular in analytical chemistry. In this work, we describe a novel DDC for capillary electrophoresis (CE) consisting of an amperometric detector (AD) and a mass spectrometer (MS). This detector combination has a good complementarity as the AD exhibits high sensitivity, whereas the MS provides excellent selectivity. Both detectors are based on a destructive detection principle, making a serial detector arrangement impossible. Thus, for the realization of the DDC, the CE flow was divided into two parts with a flow splitter. The DDC was characterized in a proof-of-concept study with ferrocene derivates and a nonaqueous background electrolyte. We could show that splitting the CE flow was a suitable method for the instrumental realization of the DDC consisting of two destructive detectors. By lowering the height of the AD compared to the MS, it was possible to synchronize the detector responses. Additionally, for the chosen model system, we confirmed that the AD was much more reproducible and had lower limits of detection (LODs) than the MS. The LODs were identical for the DDC and the single-detection arrangements, indicating no sensitivity decrease due to the CE flow splitting. The DDC was successfully applied to determine the drug and doping agent trimetazidine.     

Magnetic field dependence of photo-CIDNP MAS NMR on photosynthetic reaction centers of Rhodobacter sphaeroides WT

Photochemically induced dynamic nuclear polarization (photo-CIDNP) is observed in frozen and quinone depleted photosynthetic reaction centers of the purple bacteria Rhodobacter sphaeroides wild type (WT) by (13)C solid-state NMR at three different magnetic fields. All light-induced signals appear to be emissive at all three fields. At 4.7 T (200 MHz proton frequency), the strongest enhancement of NMR signals is observed, which is more than 10 000 above the Boltzmann polarization. At higher fields, the enhancement factor decreases. At 17.6 T, the enhancement factor is about 60. The field dependence of the enhancement appears to be the same for all nuclei. The observed field dependence is in line with simulations that assume two competing mechanisms of polarization transfer from electrons to nuclei, three-spin mixing (TSM) and differential decay (DD). These simulations indicate a ratio of the electron spin density on the special pair cofactors is 3:2 in favor of the L-BChl during the radical cation state. The good agreement of simulations with the experiments raises expectations that artificial solid reaction centers can be tuned to show photo-CIDNP in the near future.     

隐花色素磁感应模型体系的研究进展

半个世纪以来,动物地磁导航得到了广泛而深入的研究,然而其背后的磁感应机理还不清楚.1978年,Schulten等提出自由基对机理可能是生物磁感应的内在机制.近年来,这一假设得到发展和完善,提出隐花色素蛋白是生物的磁感应分子的观点.由于生物体内环境的复杂性和隐花色素蛋白的易光解特性,生物磁感应自由基对机理的研究受到很大限制.为了深入研究生物磁感应自由基对机理,人们构建了磁感应模型体系,大大简化了生物环境的复杂性,从不同方面开展了磁感应研究.基于国内外学者的研究成果,主要介绍隐花色素磁感应和各种模型体系相关研究进展,简要总结并提出生物磁感应自由基对机理研究中亟待解决的问题.     

15N Photo-CIDNP MAS NMR To Reveal Functional Heterogeneity in Electron Donor of Different Plant Organisms

In plants and cyanobacteria, two light-driven electron pumps, photosystems I and II (PSI, PSII), facilitate electron transfer from water to carbon dioxide with quantum efficiency close to unity. While similar in structure and function, the reaction centers of PSI and PSII operate at widely different potentials with PSI being the strongest reducing agent known in living nature. Photochemically induced dynamic nuclear polarization (photo-CIDNP) in magic-angle spinning (MAS) nuclear magnetic resonance (NMR) measurements provides direct excess to the heart of large photosynthetic complexes (A. Diller, Alia, E. Roy, P. Gast, H.J. van Gorkom, J. Zaanen, H.J.M. de Groot, C. Glaubitz, J. Matysik, Photosynth. Res. 84, 303–308, 2005; Alia, E. Roy, P. Gast, H.J. van Gorkom, H.J.M. de Groot, G. Jeschke, J. Matysik, J. Am. Chem. Soc. 126, 12819–12826, 2004). By combining the dramatic signal increase obtained from the solid-state photo-CIDNP effect with ¹⁵N isotope labeling of PSI, we were able to map the electron spin density in the active cofactors of PSI and study primary charge separation at atomic level. We compare data obtained from two different PSI proteins, one from spinach (Spinacia oleracea) and other from the aquatic plant duckweed (Spirodella oligorrhiza). Results demonstrate a large flexibility of the PSI in terms of its electronic architecture while their electronic ground states are strictly conserved.     

Exploring Chromophore-Binding Pocket: High-Resolution Solid-State 1H–13C Interfacial Correlation NMR Spectra with Windowed PMLG Scheme

High-resolution two-dimensional (2D) ¹H–¹³C heteronuclear correlation spectra are recorded for selective observation of interfacial 3–5.5 ? contacts of the uniformly ¹³C-labeled phycocyanobilin (PCB) chromophore with its unlabeled binding pocket. The experiment is based on a medium- and long-distance heteronuclear correlation (MELODI–HETCOR) method. For improving ¹H spectral resolution, a windowed phase-modulated Lee–Goldburg (wPMLG) decoupling scheme is applied during the t ₁ evolution period. Our approach allows for identification of chromophore–protein interactions, in particular for elucidation of the hydrogen-bonding networks and charge distributions within the chromophore-binding pocket. The resulting pulse sequence is tested on the cyanobacterial (Cph1) phytochrome sensory module (residues 1–514, Cph1Δ2) containing uniformly ¹³C- and ¹⁵N-labeled PCB chromophore (u-[¹³C,¹⁵N]-PCB-Cph1Δ2) at 17.6 T.     

Role of the Protein Cavity in Phytochrome Chromoprotein Assembly and Double-bond Isomerization: A Comparison with Model Compounds

Difference patterns of ¹³C NMR chemicals shifts for the protonation of a free model compound in organic solution, as reported in the literature (M. Stanek, K. Grubmayr [1998] Chem. Eur. J. 4 , 1653–1659), were compared with changes in the protonation state occurring during holophytochrome assembly from phycocyanobilin (PCB) and the apoprotein. Both processes induce identical changes in the NMR signals, indicating that the assembly process is linked to protonation of the chromophore, yielding a cationic cofactor in a heterogeneous, quasi-liquid protein environment. The identity of both difference patterns implies that the protonation of a model compound in solution causes a partial stretching of the geometry of the macrocycle as found in the protein. In fact, the similarity of the difference pattern within the bilin family for identical chemical transformations represents a basis for future theoretical analysis. On the other hand, the change of the ¹³C NMR chemical shift pattern upon the Pr → Pfr photoisomerization is very different to that of the free model compound upon ZZZ → ZZE photoisomerization. Hence, the character of the double-bond isomerization in phytochrome is essentially different from that of a classical photoinduced double-bond isomerization, emphasizing the role of the protein environment in the modulation of this light-induced process.