Voltamperometric determination of kinetin with a carbon paste modified electrode

The electrochemical behaviour of kinetin (6-furfurylaminopurine) on a carbon paste modified with OV-17 silicone electrode, is studied. The determination of kinetin is possible working in square wave voltammetric techniques, reaching limits of determination of 38.7 ng ml⁻¹. The proposed method was successfully applied to determine the cytokinin in extracts of apples (previously spiked with kinetin) and the obtained results were in accordance with the results obtained with HPLC-UV.     

Relation between structure and enthalpy for stacking interactions of aromatic molecules

An approximately linear correlation has been found between the enthalpy of complexation and the area of overlap of the chromophores using published structural and thermodynamical data on the self- and hetero-association of aromatic molecules measured under similar solution conditions. This finding is consistent with the assumption that short-range van-der-Waals forces dominate over other contributions to the enthalpy of stacking of aromatic molecules. It provides a ‘model-independent’ approach for a priori estimation of the enthalpy of aromatic–aromatic stacking interactions from knowledge of the structural properties or vice versa.     

Polarographic determination of glycine with a dropping copper amalgam electrode

The anodic oxidation of a dropping copper amalgam electrode in presence of dilute solutions of glycine in 0.50 M NaClO₄ has been studied. An anodic wave at ?0.28V (SCE) is observed, yielded by diffusion of glycinate anion in the solution towards the electrode surface. The wave-heights increase with the glycinate concentration (function of glycine concentration and pH value) until the anodic oxidation is controlled by the metal diffusion into mercury. The effect of pH is interpreted by attributing it to the depolarizer effects at glycinate anion even though the zwitterion is present in much larger concentrations. The applicability of anodic oxidation of a dropping copper amalgam electrode in the determination of glycine in the range of concentrations 10^?4–10^?2M with a rigorous control of pH (8.0<pH<10.5) is shown. The standard deviation of the proposed method is 4.1% and the minimum concentration determinable is in the 1×10^?4M range.     

Determination of fat-soluble vitamins in yogurt by HPLC with electrochemical detection

A method employing HPLC with electrochemical detection for the rapid and simultaneous determination of vitamins A, D(3) and E is described. The method uses a C-18 reverse phase column and 2.5 mM HAcO-NaAcO in methanol-water (99:1, v/v) solution as the mobile phase. The compounds are quantified using amperometric detection with a glassy carbon electrode at a potential of + 1300 mV (vs. Ag/AgCl) and the results are compared with those obtained using UV detection at a wavelength of 280 nm. The method was successfully applied to the analysis of vitamins A, D(3) and E in yogurt samples. After saponification, fat-soluble vitamins were extracted and the methanolic solution of the extracts was injected directly into the chromatographic system, avoiding the clean-up step which is necessary when no electrochemical detection is used. Good recovery percentages were obtained.     

Structural characterization of Acetobacter diazotropicus levansucrase by matrix-assisted laser desorption/ionization mass spectrometry: identification of an N-terminal blocking group and a free-thiol cysteine residue

High-resolution matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to characterize the primary structure of the levansucrase (EC 2.4.1.10) secreted by Acetobacter diazotropicus SRT4. The technique permitted not only the reading frame of this enzyme, the amino acid sequence of which was deduced from DNA, but also the elucidation of an N-terminal blocking group and the position of a disulfide bridge between Cys309 and Cys365 among the three Cys residues. A free cysteine (Cys127) was identified by modifying an intact molecule with a sulfhydryl reagent, 5-(octyldithio)-2-nitrobenzoic acid, under non-reducing conditions. In addition, the enzyme obtained by site-directed mutagenesis at Asp279 to Asn279 was also identified by the above methods. Post-source decay analysis of the tryptic peptide containing the mutation site unequivocally revealed an Asn residue at position 279.     

Use of a partial-filling technique in affinity capillary electrophoresis for determining binding constants of ligands to receptors

This work evaluates the concept of a partial-filling technique in affinity capillary electrophoresis (ACE) using two model systems: vancomycin from Streptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). In this technique the capillary is first partially-filled with ligand followed by a sample of receptor and non-interacting standard and electrophoresed. Analysis of the change in the mobility ratio, M, of the receptor, relative to the non-interacting standard, as a function of the concentration of the ligand, yields a value for the binding constant. These values agree well with those estimated using other binding and ACE techniques. Data demonstrating the quantitative potential of this method is presented.