粗糙化银电极上氰基钴胺素的电化学行为

比较研究氰基钴胺素在光滑和粗糙化银电极上的电化学行为，发现在光滑银电极上氰基钴胺素的还原为一个二电子过程，而在粗糙化银电极上则为两个一电子过程，其原因是由于粗糙化银电极对氰根的强吸附作用。     

A novel poly(cyanocobalamin) modified glassy carbon electrode as electrochemical sensor for voltammetric determination of peroxynitrite

This report described the direct voltammetric detection of peroxynitrite (ONOO⁻) at a novel cyanocobalamin modified glassy carbon electrode prepared by electropolymeriation method. The electrochemical behaviors of peroxynitrite at the modified electrode were studied by cyclic voltammetry. The results showed that this new electrochemical sensor exhibited an excellent electrocatalytic activity to oxidation of peroxynitrite. The mechanism of catalysis was discussed. Based on electrocatalytic oxidation of peroxynitrite at the poly(cyanocobalamin) modified electrode, peroxynitrite was sensitively detected by differential pulse voltammetry. Under optimum conditions, the anodic peak current was linear to concentration of peroxynitrite in the range of 2.0 × 10⁻⁶ to 3.0 × 10⁻⁴ mol L⁻¹ with a detection limit of 1.0 × 10⁻⁷ mol L⁻¹ (S/N of 3). The proposed method has been applied to determination of peroxynitrite in human serum with satisfactory results. This poly(cyanocobalamin) modified electrode showed high selectivity and sensitivity to peroxynitrite determination, which could be used in quantitative detection of peroxynitrite in vivo and in vitro.     

Investigation of Cyanocobalamin Interferences to an Electrochemical Based Hemoglobin Test System

The structure of vitamin B-12 with a cobalt ion in the center is similar to the structure of heme with an iron center in the hemoglobin. The suspicion that the presence of vitamin B-12 in venous blood samples caused interference in readings of hemoglobin levels when using the BeneCheck hemoglobin test system, an electrochemical based strip and meter, was investigated in this study. Venous blood samples with 5 mg/dL of vitamin B-12 induced a positive error of 13% in the hemoglobin measurement, while 10 mg/dL of vitamin B-12 caused a positive error of 30% in the hemoglobin measurement. However, 1 mg/dL of vitamin B-12, a concentration much higher than that of venous blood concentrations in patients under B-12 treatment, did not cause an error in the hemoglobin measurement. These results suggest that the BeneCheck hemoglobin test system is suitable for clinical application for patients under B-12 treatment because the dosage is below 1 mg/dL.     

Modulation of mobile phase composition in flow-injection/sequential-injection chromatography exploiting multisyringe flow analysis

In this paper, a time-based multicommutated flow system is proposed for appropriate selection and modulation of mobile phase composition in flow-injection (FI)/sequential-injection (SI) chromatography. The novel flow assembly involves the on-line coupling of a short monolithic reversed-phase chromatographic column with a multisyringe flow injection set-up furnished with a set of solenoid valves. The proposed hyphenated technique was applied to the simultaneous spectrophotometric determination of thiamine (B₁), pyridoxine (B₆) and cyanocobalamin (B₁₂) which were taken as model analytes. The separation method capitalizes on a dual isocratic elution protocol involving the use of a single forward stroke of the multisyringe pump for initial delivery of 50 mmol L⁻¹ ammonium acetate (pH 7.0) for 2.4 min followed by 50 mmol L⁻¹ ammonium acetate–methanol (80:20, v/v) for 6.4 min at 0.5 mL min⁻¹ and room temperature. Detection was performed at the maximum wavelength for each target vitamin—280 nm for B₁, 325 nm for B₆, and 360 nm for B₁₂. A first-order, two-level full-factorial design was utilized to ascertain the significant variables influencing the chromatographic separation and the magnitude of the interaction effects. The experimental design method revealed that resolution of the target vitamins is highly dependent on the pH, percentage of organic modifier, and their second-order interaction. The multisyringe flow-injection-based monolithic column separation method, which should be viewed as an expeditious and cost-effective alternative to the high-performance liquid chromatography counterpart, was applied to the separation and determination of B₁, B₆, and B₁₂ in different pharmaceutical dosage forms in less than 9 min. Statistical comparison of the results from the proposed procedure with those from the HPLC method endorsed by the US Pharmacopeia revealed there were no significant differences at the 95 % confidence level.     

Spektralphotometrische Bestimmung von Kobalt mit 2-(5-Brom-2-pyridylazo)-5-Diethylaminophenol

Zusammenfassung Eine empfindliche und selektive spektralphotometrische Kobaltbestimmung (<0,9 g ml^–1) ist mit 2-(5-Brom-2-pyridylazo)-5-diethylaminophenol (5-Brom-PADAP) auf Grund des kinetisch stabilen Co(III)L ₂ ^– -Chelates möglich, das nach seiner Bildung bei pH 7 (Ammoniumacetat) in Gegenwart von 0,1% Triton X-100, 5,10^–3 M Ammoniumperoxodisulfat und 10% (v/v) Dimethylformamid (auch nach Zugabe von 1,25 M H₂SO₄, 1,80 M HNO₃ oder auch 0,1–0,01 M EDTA) stabil ist. Eine Reihe von 5-Brom-PADAP-Chelaten störender Ionen werden dabei zersetzt. V(V), Hg²⁺, Ni²⁺, Cu²⁺, Pd²⁺ können stören. Die Kobaltbestimmung in Cyanocobalamin und auch Trinkwasser ist auf diesem Wege möglich. Die Fehler lagen im Bereich von — 3 bis — 7 % (1,8–9 g Co/l).Herrn Prof. Dr. F. Umland, Westfälische Universität, Münster/Westfalen, sind wir für das 5-Brom-PADAP mit herzlichem Dank verpflichtet.     

Excited electronic states and internal conversion in cyanocobalamin

Cyanocobalamin (CNCbl) is a paradigm system for the study of excited electronic states and biological cofactors including the B12 vitamers. The photophysics of CNCbl has been thoroughly investigated using both ultrafast spectroscopy and time dependent density functional theory (TD-DFT). Herewe review the spectroscopic and theoretical investigations of CNCbl with emphasis on the nature of S1, the lowest excited electronic state, and extend the spectroscopic measurements to include the ultraviolet region of the spectrum. Ultrafast transient absorption measurements in the visible αβ band region and in the midinfrared led to assignment of the S1 state to a ligand-to-metal charge transfer (LMCT) with lengthened axial bonds and a ~3 kcal/mol barrier for internal conversion to the ground state. The present measurements encompassing the γ band region of the spectrum provide further support for the assignment of the S1 state. The experiments are in good agreement with the results of TD-DFT calculations which confirm the expected lengthening of the axial bonds in S1 and account for the observed barrier for internal conversion back to the ground state.