María Cuartero

Assay of acetylcholinesterase activity by potentiometric monitoring of acetylcholine

An acetylcholine-selective electrode based on a plasticized polymeric membrane has been developed. The electrode exhibited good selectivity for acetylcholine (ACh) over choline and some common ions, low drift, and a fast response to ACh. The response was linear over an ACh concentration range of 1×10(-6) to 1×10(-3) M with a slope of 59.1±0.1 and a detection limit of 1.5×10(-7)±1.2×10(-8) M. The electrode was used to monitor enzymatic ACh hydrolysis catalyzed by acetylcholinesterase (AChE) at different substrate and enzyme concentrations. A kinetic data analysis permitted the determination of the Michaelis-Menten constant of the enzymatic hydrolysis and AChE activity in the range of 2×10(-5) to 3.8×10(-1)U ml(-1).

Rubber-based substrates modified with carbon nanotubes inks to build flexible electrochemical sensors

The development of a solid-contact potentiometric sensor based on conducting rubbers using a carbon nanotubes ink is described here. Commercial rubbers are turned into conductive ones by a simple and versatile method, i.e. painting an aqueous dispersion of single-walled carbon nanotubes on the polymer surface. On this substrate, both the working ion-selective electrode and the reference electrode are built in order to form an integrated potentiometric cell. As a proof-of-principle, selective potassium electrodes are fully characterized giving comparable performances to conventional electrodes (sensitivity, selectivity, stability, linear range, limit of detection and reproducibility). As an application of the rubber-based electrodes, a bracelet was constructed to measure potassium levels in artificial sweat. Since rubbers are ubiquitous in our quotidian life, this approach offers great promise for the generation of chemical information through daily objects.

New approach for the potentiometric-enzymatic assay of reversible-competitive enzyme inhibitors. Application to acetylcholinesterase inhibitor galantamine and its determination in pharmaceuticals and human urine

A new kinetic-potentiometric method for the characterization and analytical determination of competitive reversible enzyme inhibitors was developed. The method is based on a mathematical approach, assuming that the reaction proceeds at the steady state, which permits calculation of a tentative substrate concentration to be used to determine low inhibitor concentrations and to obtain the value of inhibition constant corresponding to the inhibitor. The mathematical approach predicts a linear relationship between the inverse of the relative inhibition and the inverse of the inhibitor concentration. The method developed is applied to the acetylcholinesterase inhibitor galantamine, using an acetylcholine-selective electrode. A linear relationship for galantamine concentration from 2×10(-8) to 1×10(-6)M and a limit of detection of 5.4×10(-9)M was found. A value for KI(Gal) of 2.0×10(-7)±0.1×10(-7)M was obtained. The effect of several other drugs and of the main galantamine metabolite excreted in urine was studied. The method was satisfactorily applied to the determination of galantamine in pharmaceuticals and human urine.

New carbazolo[1,2-a]carbazole derivative as ionophore for anion-selective electrodes: remarkable recognition towards dicarboxylate anions.

A new carbazolo[1,2-a]carbazole derivative was synthesized by expanding the binding cavity to explore the possibility of hosting larger anions such as dicarboxylate anions. The compound was incorporated as an ionophore into a membrane for an anion-selective electrode. The response of the electrode was evaluated for oxalate, malonate, succinate, glutarate and adipate in terms of calibration characteristics (slope, limit of detection and linear range of the response), response time, repeatability, reproducibility and selectivity. Nernstian reproducible responses, with very good detection limits, fast responses and selectivity not previously observed, were found for all the dicarboxylates anions, and the results were especially good in the case of glutarate. In order to obtain additional structural information about the complex formed between the ionophore and the dicarboxylate anions, (1)H NMR and fluorescence studies were carried out. The observed potentiometric selectivity depends on the good correspondence between the size of the carbazolocarbazole cavity and the length of the dicarboxylate anion, as supported by the NMR and fluorescence studies.

A SO2-selective electrode based on a Zn-porphyrin for wine analysis.

This work describes the assessment of a SO2-selective electrode based on the use of the neutral carrier 5,10,15,20-tetraphenyl(porphyrinate)zinc(II) in a PVC membrane plasticized with 2-nitrophenyl phenyl ether. After being conditioned in 2 mol L(-1) diethylamine solution for 24 h, the electrode exhibited selective anionic response toward the analyte in a concentration interval of more than four decades, with an slope of -59.5 mV dec(-1), a practical detection limit of 3.7×10(-6) mol L(-1) and a low limit of linear range of 7.2×10(-6) mol L(-1). The response mechanism is based on the displacement of the diethylamine:metalloporphyrin complex equilibrium within membrane bulk, inducing a variation in the cationic-sites to ionophore ratio. In turn, free hydroxyl ions are complexed by the displaced ionophore in a ratio 1:1 and translated as single negative charge nernstian response. Finally, the selectivity of the electrode is evaluated in view of its application to wine analysis. Results had high accuracy and precision when compared with a reference method.

Benzodipyrrole derivates as new ionophores for anion-selective electrodes: Improving potentiometric selectivity towards divalent anions

Two open substituted benzodipyrroles were tested as hydrogen-bond-forming anion ionophores for the development of anion-selective electrodes. These compounds were incorporated in plasticized polymeric membranes with different plasticizers, using different membrane compositions to explore their response towards several anions. The electrodes constructed with membranes containing 2-nitrophenyl octyl ether and a 0.5 molar ratio ionic additive/ionophore showed pronounced anti-Hofmeister behaviour, providing a significantly enhanced response towards the divalent anions sulfate, sulfite, thiosulfate and oxalate. The selected electrodes were also evaluated in terms of detection limits and selectivity. (1)H NMR experiments were carried out in an attempt to explain some aspects of the behaviour observed.

Terminal carbohydrates abundance, immune related enzymes, bactericidal activity and physico-chemical parameters of the Senegalese sole (Solea senegalensis, Kaup) skin mucus

ABSTRACT Recently, interest in mucosal surfaces, more specifically fish skin and its secreted mucus, has greatly increased among immunologists. The abundance of terminal carbohydrates, several enzymes (proteases, lysozyme, peroxidase, alkaline phosphatase, esterases and ceruloplasmin), bactericidal activity against fish pathogenic and non‐pathogenic bacteria and several physico‐chemical parameters (protein concentration, pH, conductivity, redox potential, osmolarity, density and viscosity) in the skin mucus of Senegalese sole (Solea senegalensis, Kaup) have been evaluated. Present results evidence the abundance of N‐acetylneuraminic acid, mannose, glucose and N‐acetyl‐galactosamine in skin mucus. The levels of lysozyme, proteases, esterases and alkaline phosphatase were very similar (from 20 to 30 Units mg−1 protein). However, 93 Units mg−1 protein were detected of ceruloplasmin and only 4′88 Units mg−1 protein of peroxidase. Skin mucus of S. senegalensis showed high bactericidal activity against the tested pathogen bacteria but weak activity against non‐pathogenic bacteria. Finally, a clear relationship between mucus density and temperature was detected, while viscosity showed a direct shear‐ and temperature‐dependent behaviour. These results could be useful for better understanding the role of the skin mucus as a key component of the innate immune system, as well as, for elucidating possible relationships between biological and physico‐chemical parameters and disease susceptibility. HIGHLIGHTSN‐acetylneuraminic acid, mannose, glucose and N‐acetyl‐galactosamine are present in skin mucus of Solea senegalensis.Levels of ceruloplasmin and peroxidase were higher and lower, respectively, than the levels of the other studied enzymes.Skin mucus showed high bactericidal activity against the tested pathogen bacteria but weak activity against non‐pathogenic bacteria.Mucus density and temperature were related. Viscosity showed a direct shear‐ and temperature‐dependent behaviour.The results could be useful for elucidating possible relationships between biological and physico‐chemical parameters.

Use of a New Ziprasidone-Selective Electrode in Mixed Solvents and Its Application in the Analysis of Pharmaceuticals and Biological Fluids

The construction and characterization of a new ion-selective electrode for the determination of the antipsychotic ziprasidone in mixed solvents is presented. The electrode contains a plasticized polymeric membrane based on a ziprasidone-tetraphenylborate ion-exchanger. The influence of membrane composition on the electrode response towards ziprasidone in hydroalcoholic solutions was studied. The electrode displayed a stable response in a 2:3 (v/v) methanol/water medium from a ziprasidone concentration of 3 × 10−6 M with a fast response time of less than 20 s. The electrode also showed good selectivity towards ziprasidone over common inorganic and organic compounds and several species with pharmacological activity. The electrode was successfully applied to the determination of ziprasidone in pharmaceuticals and human urine and serum.