Comparison of biosensors based on gold and nanocomposite electrodes for monitoring of malic acid in wine
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Abstract Amperometric biosensors based on a gold planar electrode and on two types of nanocomposite electrodes consisting of multi-walled carbon nanotubes for the determination of L-malic acid designed for wine-makers were developed. The biosensors designed for wine-makers were constructed by immobilization of L-malate dehydrogenase and diaphorase within chitosan layers on the surface of the electrodes. The coenzyme NAD+ and the electrochemical mediator ferricyanide were present in the measuring solution. The current resulting from re-oxidation of produced ferrocyanide was measured at a working potential of +300 mV against an Ag/AgCl reference electrode. The biosensor based on a gold electrode showed linearity over the range 10–520 µM with a detection limit of 5.41 µM. Calibration curves for biosensors utilizing nanocomposites were obtained both with the linear range of 10 to 610 µM. The detection limits were 1.57 and 1.77 µM, respectively. The biosensors showed satisfactory operational stability (no loss of sensitivity after 30 consecutive measurements) and storage stability (90% of the initial sensitivity after one year of storage at room temperature). The results obtained from measurements of wine samples were in a good correlation with the standard HPLC method. Satisfactory biosensor sensitivity, specificity and stability allowed their successful commercialization.Keywords:
Amperometry
Ferrocyanide
Ferricyanide
Prussian blue
Malic acid
Prussian blue
Ferricyanide
Ferrocyanide
Potassium ferricyanide
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Fourier transform infrared (FT-IR) spectrometry adds complementary structural information to sensitive electroanalytical measurements. In this work, in situ, external reflection FT-IR techniques allowed for examination of interfacial electrochemical events through a thin-layer spectroelectrochemical cell in which a sinusoidally modulated potential was applied to the electrode. The infrared spectra of electrochemically modulated species were measured by a step-scan FT-IR spectrometer. The phase-sensitive detection capability of this technique has been used to provide information about dynamic electrode processes and to enhance sensitivity to surface species. The cause of decreases in the heterogeneous rate constant, k°, of the ferricyanide/ferrocyanide redox process with time has been investigated. Previous authors offer a variety of opinions for the variation of k° for this reaction including the adsorption of polymeric hexacyanoferrate complexes, ferrocyanide, or ferricyanide on the electrode surface; other reports indicate that no species adsorb on the electrode during the electron-transfer process. Our results indicate that, with KCl as electrolyte, a hexacyanoferrate complex can form during potential cycling in the ferri-/ferrocyanide redox couple, but only under rather extreme experimental parameters. Measurements made by electrochemical modulation combined with step-scan FT-IR spectrometry show that, under other conditions, the change in rate constant of this redox process can be attributed to the adsorption of ferrocyanide ion.
Ferrocyanide
Ferricyanide
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1. Reduction of ferricyanide by the isolated perfused rat liver and by isolated rat hepatocytes was studied. 2. Ferricyanide was reduced to ferrocyanide by the perfused liver at a linear rate of 0.22μmol/min per g of liver. Ferricyanide was not taken up by the liver and the perfusate concentration of ferricyanide+ferrocyanide remained constant throughout the perfusion. Perfusate samples from livers perfused without ferricyanide did not reduce ferricyanide. 3. Isolated hepatocytes reduced ferricyanide in a biphasic manner. The initial rate of 2.3μmol/min per g of cells proceeded for approx. 3min and derived from low-affinity sites (apparent Km>1.3mm). The secondary rate of 0.29μmol/min per g of cells was maintained for the remainder of the incubation and derived from higher affinity sites (apparent Km0.13mm). Disruption of the cells resulted in an increase in the low-affinity rate and a decrease in the high-affinity rate. 4. Ferrocyanide was oxidized by isolated hepatocytes but not by perfused liver. The apparent Km for ferrocyanide oxidation by hepatocytes was 1.3mm. 5. Oxidized cytochrome c was reduced by isolated hepatocytes in the presence of 1mm-KCN but at a rate less than that of the reduction of ferricyanide. 6. Properties of the ferricyanide-reducing activities of intact hepatocytes and the perfused liver were examined. The low-affinity rate, present only in cell and broken cell preparations, was inhibited by 1μm-rotenone and 0.5mm-ferrocyanide, and stimulated by 0.1mm-KCN. The mitochondrial substrate, succinate, also stimulated this rate. The perfused liver showed only a high-affinity activity for ferricyanide reduction. This activity was also present in liver cells and was unaffected by rotenone, antimycin A, KCN, NaN3, or p-hydroxymercuribenzoate but was inhibited by 2.6mm-CaCl2, 2-heptyl-4-hydroxyquinoline-N-oxide and ferrocyanide. Overall, these results are consistent with the occurrence of a trans-plasma membrane redox system of liver that reduces extracellular ferricyanide to ferrocyanide. The reduction process shows properties which are similar to that of the NADH:ferricyanide oxidoreductase found in isolated liver plasma membranes but different from that of mitochondria.
Ferricyanide
Ferrocyanide
Potassium ferricyanide
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Prussian blue
Ferricyanide
Ferrocyanide
Potassium ferricyanide
Amperometry
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Prussian blue
Ferrocyanide
Ferricyanide
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Ferrocyanide
Ferricyanide
Potassium ferricyanide
Prussian blue
Potassium ferrocyanide
Potassium hydroxide
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The hypothesis is tested that acidification of the bulk medium during transplasmalemma electron transport to ferricyanide is due solely to a requirement for charge balance. According to this hypothesis, reduction of the trivalent anion, ferricyanide, to the tetravalent anion, ferrocyanide, results in a charge difference that is balanced by protons. A coulometric device is used that rapidly and efficiently reoxidizes ferrocyanide to ferricyanide, thus maintaining a constant charge in the bulk medium. Oat (Avena sativa L. cv Garry) mesophyll protoplasts are chosen as experimental material to facilitate ferricyanide reduction and the concomitant ferrocyanide reoxidation by the coulometric device. The kinetics of ferricyanide reduction and proton excretion by protoplasts are similar to those of other cell types and tissues. Rates of net proton excretion are identical regardless of whether the ferrocyanide is simultaneously reoxidized. We conclude that acidification may occur during transplasmalemma electron transport when there is no change in negative charge of the bulk medium.
Ferricyanide
Ferrocyanide
Protoplast
Coulometry
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Ferricyanide
Ferrocyanide
Chemisorption
Surface-Enhanced Raman Spectroscopy
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Ferrocyanide
Ferricyanide
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Prussian Blue(PB) film and copper ferricyanide composite with PB were prepared in our laboratory. Thire electrochemical properties were investigated with cyclic voltammetry and in situ spectroelectrochemistry. The results depicted that Prussian Blue film and its composite film both exhibited excellent electrochemical features with good stability. Therefore, they could be acted as a potential candidate for electrochemical display device.
Prussian blue
Ferricyanide
Composite film
Ferrocyanide
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