RESUMO
This work reports a new method to evaluate the antioxidant capacity of infusions and beverages, based on superoxide radicals. Radicals produced by the enzymatic reaction between acetylcholinesterase and hypoxanthine oxidized antioxidant molecules present in commercially available samples or standard solutions, which was monitored by means of cyclic voltammetry using a carbon paste electrode. The Trolox equivalent antioxidant capacity (TEAC) of red wine, coffee and green tea determined using this method were: (1.20 ± 0.06), (0.90 ± 0.02), and (0.65 ± 0.02), respectively. This method suggested TEACred wine > TEACcoffee > TEACgreen tea, which is the same as DPPH, spectrophotometric method. However, the electrochemical one proposed here is rapid and simple.
Assuntos
Antioxidantes/química , Bebidas/análise , Técnicas Eletroquímicas/métodos , Superóxidos/química , Antioxidantes/metabolismo , Café/química , Eletrodos , Concentração de Íons de Hidrogênio , Hipoxantina/química , Hipoxantina/metabolismo , Oxirredução , Superóxidos/metabolismo , Chá/química , Vinho/análise , Xantina Oxidase/metabolismoRESUMO
The working area of a screen-printed electrode, SPE, was modified with the enzyme tyrosinase (Tyr) using different immobilization methods, namely entrapment with water-soluble polyvinyl alcohol (PVA), cross-linking using glutaraldehyde (GA), and cross-linking using GA and human serum albumin (HSA); the resulting electrodes were termed SPE/Tyr/PVA, SPE/Tyr/GA and SPE/Tyr/HSA/GA, respectively. These biosensors were characterized by means of amperometry and EIS techniques. From amperometric evaluations, the apparent Michaelis-Menten constant, Km', of each biosensor was evaluated while the respective charge transfer resistance, Rct, was assessed from impedance measurements. It was found that the SPE/Tyr/GA had the smallest Km' (57 ± 7) µM and Rct values. This electrode also displayed both the lowest detection and quantification limits for catechol quantification. Using the SPE/Tyr/GA, the Trolox Equivalent Antioxidant Capacity (TEAC) was determined from infusions prepared with "mirto" (Salvia microphylla), "hHierba dulce" (Lippia dulcis) and "salve real" (Lippia alba), medicinal plants commonly used in Mexico.
Assuntos
Antioxidantes/química , Técnicas Biossensoriais/métodos , Plantas Medicinais/química , Catecóis/química , Eletroquímica/métodos , Eletrodos , Enzimas Imobilizadas/química , Humanos , México , Monofenol Mono-Oxigenase/químicaRESUMO
This work deals with the formation of supramolecular complexes between ascorbic acid (AA), the guest, and ß-cyclodextrin (ß-CD), the host, that was first potentiodynamically immobilized on the surface of a carbon paste electrode (CPE) throughout the formation of a ß-CD-based conducting polymer (poly-ß-CD). With the bare CPE and the ß-CD-modified CPE, an electrochemical study was performed to understand the effect of such surface modification on the electrochemical response of the AA. From this study it was shown that on the modified-CPE, the AA was surface-immobilized through formation of an inclusion complex with ß-CD, which provoked the adsorption of AA in such a way that this stage became the limiting step for the electrochemical oxidation of AA. Moreover, from the analysis of the experimental voltammetric plots recorded during AA oxidation on the CPE/poly-ß-CD electrode surfaces, the Gibbs' standard free energy of the inclusion complex formed by the oxidation product of AA and ß-CD has been determined for the first time, ∆G0inclus = -36.4 kJ/mol.
Assuntos
Ácido Ascórbico/química , beta-Ciclodextrinas/química , Carbono/química , Eletroquímica , Eletrodos , Concentração de Íons de Hidrogênio , Imobilização , Oxirredução , Propriedades de SuperfícieRESUMO
Four flavonols, namely quercetin, morin, kaempferol, and myricetin, were studied using spectrophotometry (UV-vis) in aqueous solution. The study was performed varying the pH to analyze the stability of these compounds, and to estimate their acidity constants. In addition, the deprotonation mechanisms were studied using computational chemistry within the density functional theory framework. The calculations were performed in aqueous solution using the SMD continuum model, and the results are reported as deprotonation energies. Our results show that both quercetin and myricetin are highly unstable at basic pH. Kaempferol, on the other hand, is much more stable, and morin is the only one among the studied compounds that was not affected by pH. In spite of this inconvenience, their acidity constants were estimated through analysis of their decomposition kinetics, correcting the spectra accordingly, and obtaining a correlation of values between the experimentally observed pKa and the calculated ΔG of successive deprotonations.
Assuntos
Flavonóis/química , Prótons , Concentração de Íons de Hidrogênio , Cinética , Soluções , Espectrofotometria Ultravioleta , Termodinâmica , Água/químicaRESUMO
An amperometric biosensor based on laccase, from Trametes versicolor (LTV), was developed and optimized for monitoring the phenolic compounds content in tea infusions. The fungal enzyme was immobilized by entrapment within polyvinyl alcohol photopolymer PVA-AWP (azide-unit pendant water-soluble photopolymer) onto disposable graphite screen-printed electrodes (SPE). Sensitivity optimization in terms of pH, temperature and applied potential was carried out. The linear range, detection limit, operational and storage stabilities were also determined. The laccase biosensor (LTV-SPE) was calibrated for o-, m- and p-diphenol as well as caffeic acid. The highest response was found at 0.1M acetate buffer pH 4.7, though it must be added the good reproducibility and operational stability were also obtained. The useful lifetime of the biosensor is estimated to be greater than 6 months. LTV-SPE was used for the determination of the equivalent phenol content (EPC) in tea infusions by the direct addition into the electrochemical cell: the results were compared with those from the Folin-Ciocalteu spectrophotometric method. The amperometric detection exhibits some interesting advantages such as high simplicity, minimal sample preparation and shorter response time. A stable and sensitive amperometric response was obtained toward standard diphenolic compounds and herbal infusions. These biosensors are useful for easy and fast monitoring of EPC that can be related to the antioxidant capacity of natural extracts.
Assuntos
Técnicas Biossensoriais , Eletroquímica/métodos , Análise de Alimentos/métodos , Fenol/química , Chá , Antioxidantes/química , Calibragem , Eletrodos , Concentração de Íons de Hidrogênio , Polímeros/química , Sensibilidade e Especificidade , Espectrofotometria/métodos , Temperatura , Água/químicaRESUMO
In this work it is explained, by the first time, the application of programs SQUAD and HYPNMR to refine equilibrium constant values through the fit of electrophoretic mobilities determined by capillary zone electrophoresis experiments, due to the mathematical isomorphism of UV-vis absorptivity coefficients, NMR chemical shifts and electrophoretic mobilities as a function of pH. Then, the pK(a) values of tenoxicam in H(2)O/DMSO 1:4 (v/v) have been obtained from (1)H NMR chemical shifts, as well as of oxicams in aqueous solution from electrophoretic mobilities determined by CZE, at 25 degrees C. These values are in very good agreement with those reported by spectrophotometric and potentiometric measurements.
Assuntos
Eletroforese Capilar/métodos , Espectroscopia de Ressonância Magnética/métodos , Piroxicam/análogos & derivados , Algoritmos , Eletroforese Capilar/instrumentação , Concentração de Íons de Hidrogênio , Cinética , Meloxicam , Modelos Químicos , Estrutura Molecular , Piroxicam/química , Software , Estereoisomerismo , Tiazinas/química , Tiazóis/químicaRESUMO
Amperometric acetylcholinesterase biosensors have been developed for quantification of the pesticides carbofuran, carbaryl, methylparaoxon, and dichlorvos in phosphate buffer containing 5% acetonitrile. Three different biosensors were built using three different acetylcholinesterase (AChE) enzymes-AChE from electric eel, and genetically engineered (B394) and wild-type (B1) AChE from Drosophila melanogaster. Enzymes were immobilized on cobalt(II) phthalocyanine-modified electrodes by entrapment in a photocrosslinkable polymer (PVA-AWP). Each biosensor was tested against the four pesticides. Good operational stability, immobilisation reproducibility, and storage stability were obtained for each biosensor. The best detection limits were obtained with the B394 enzyme for dichlorvos and methylparaoxon (9.6 x 10(-11) and 2.7 x 10(-9) mol L(-1), respectively), the B1 enzyme for carbofuran (4.5 x 10(-9) mol L(-1)), and both the B1 enzyme and the AChE from electric eel for carbaryl (1.6 x 10(-7) mol L(-1)). Finally, the biosensors were used for the direct detection of the pesticides in spiked apple samples.
Assuntos
Acetonitrilas , Acetilcolinesterase/metabolismo , Técnicas Biossensoriais/métodos , Carbaril/análise , Carbofurano/análise , Diclorvós/análise , Paraoxon/análogos & derivados , Animais , Carbaril/metabolismo , Carbofurano/metabolismo , Diclorvós/metabolismo , Drosophila melanogaster/enzimologia , Electrophorus/metabolismo , Malus/química , Estrutura Molecular , Paraoxon/análise , Paraoxon/metabolismo , Praguicidas/análise , Praguicidas/metabolismoRESUMO
This paper presents the construction of an amperometric biosensor for the highly sensitive detection of the organophosphorus insecticide dichlorvos, based on the inhibition of acetylcholinesterase (AChE). The sensitivity of three AChEs from different sources were tested and compared: AChEs from Electric eel (Ee) and genetically engineered (B394) and wild type (B1) from Drosophila melanogaster (Dm). The enzymes were immobilized by entrapment in a photocrosslinkable PVA-SbQ polymer on a screen printed graphite electrode. The enzyme activity was estimated amperometrically at 100mV versus Ag/AgCl by measuring the thiocholine produced by the enzymatic hydrolysis of the acetylthiocholine substrate using cobalt phthalocyanine as electron mediator. The pesticide was measured in the presence of 5% acetonitrile without loss of enzyme activity. The best sensitivity was achieved with the Dm mutant B394 with a detection limit of 7x10(-11)M as compared to 1x10(-8)M with the B1 Dm and 6x10(-7)M with the Ee. The B394 biosensor was used to quantify dichlorvos in a sample of skin apple after extraction with acetonitrile.
Assuntos
Técnicas Biossensoriais , Inibidores da Colinesterase/análise , Diclorvós/análise , Eletroquímica/métodos , Malus/química , Resíduos de Praguicidas/análise , Acetilcolinesterase/metabolismo , Calibragem , Sensibilidade e EspecificidadeRESUMO
This work presents the results obtained from a spectrophotometry study performed on some indicators of the sulfonphtaleins like phenol red (PR), thymol blue (TB), bromothymol blue (BTB), xylenol orange (XO) and methylthymol blue (MTB). During the first stage the acidity constants of some of the indicators were determined using the data from spectrophotometry, potentiometry and with the use of the software SQUAD. These were as follows: for the equilibrium 2H+BTB<-->H(2)BTB, log beta(2)=15.069+/-0.046 and for H+BTB<-->HBTB, log beta(1)=8.311+/-0.044. For the XO and the MTB five values were calculated for each, namely, for MTB: log beta(5)=42.035, log beta(4)=38.567+/-0.058, log beta(3)=32.257+/-0.057, log beta(2)=23.785+/-0.057, and log beta(1)=12.974+/-0.045 while for XO: log beta(5)=40.120+/-0.102, log beta(4)=35.158+/-0.062, log beta(3)=29.102+/-0.053, log beta(2)=21.237+/-0.044, and log beta(1)=11.682+/-0.044. During the second stage, a study was conducted on the effect of the substituents present in the indicators to determine the effect of different functional groups on the pK(a) value corresponding to the last indicator's dissociation.
Assuntos
Indicadores e Reagentes/química , Azul de Bromotimol/análogos & derivados , Azul de Bromotimol/química , Simulação por Computador , Concentração de Íons de Hidrogênio , Modelos Químicos , Fenóis , Fenolsulfonaftaleína/química , Prótons , Espectrofotometria Ultravioleta , Sulfóxidos , Timolftaleína/análogos & derivados , Timolftaleína/química , Xilenos/químicaRESUMO
The manufacture and evaluation of a novel sensor built with a composite material, highly selective to nitrate ions using doped polypyrrole as a recognition agent, are presented. When the ratio of recognition agent to graphite was optimized at 1:1, and the sensitivities found closely approached nernstian behavior. The stability times attained were less than 14 min with response times also below 20 s. Batch characterization of the sensor displayed a sensitivity of 57.1 mV/decade of nitrate ion activity (alphaNO3-) and a detection limit of 5.37 x 10(-5) M, which are comparable to those reported for commercial sensors. Evaluation of the selectivity coefficients showed high affinity to nitrate ion, superior to that of commercial sensors and others reported in the literature. The composite material gives the sensor a prolonged service life with the added capability of allowing the regeneration of its active surface. Coupling the sensor and a solid state, composite-type, reference electrode to a flow injection analysis system (FIA) permitted to achieve an effective overall assessment of the system. A nitrate determination test was conducted on real samples. A comparison of the results obtained, either with stationary measurements or with FIA, indicated that there were no significant differences from the values from manufacturer's specifications.
Assuntos
Nitratos/análise , Nitratos/química , Polímeros/química , Pirróis/química , Calibragem , Concentração de Íons de Hidrogênio , Dinitrato de Isossorbida/análogos & derivados , Dinitrato de Isossorbida/química , Sensibilidade e EspecificidadeRESUMO
The equilibrium constants of salicylaldoxime in water at 25 degrees C, 0.5 M of ionic strength with NaCl and concentration of 1x10(-4) M were determined spectrophotometrically. The spectral data were processed using SQUAD program. The salicylaldoxime in acid medium has the value of pKa1=1.224+/-0.027. In alkaline medium the salicylaldoxime has the values of pKa2=8.551+/-0.024 and pKa3=11.728+/-0.016.
Assuntos
Ácido Clorídrico/química , Oximas/análise , Cloreto de Sódio/química , Temperatura , Concentração de Íons de Hidrogênio , Cinética , Concentração Osmolar , Oximas/química , Soluções , Espectrofotometria UltravioletaRESUMO
This work presents the results concerning the first use of activated composite membranes (ACMs) for the removal of Hg(II) ions from aqueous solution, using as the ligand di-(2-ethylhexyl)dithiophosphoric acid (DTPA). The effects on the removal percentage of Hg(ll) of variables such as pH, the nature of the acid, the concentration of mercury (in the feed solutions), and the ligand content (in the membrane) as well as the total surface membrane area exposed to the Hg(II) aqueous solution were studied. During the course of the removal experiments, the membrane was immersed in the Hg(II) aqueous solution in acid media and samples of the solution were taken at different times to enable monitoring of the mercury concentration changes. Itwasfound that when the ACM was prepared with a 1.0 M DTPA casting solution and the feed solution contained 2.49 x 10(-4) M Hg(II) in HCI 0.1 M the amount of mercury extracted was higher than 93%. Straight forwardly, additional experiments were carried out with the free-DTPA composite membranes to make up a set of control reference points to verify that removal of the investigated heavy metal was a consequence of the presence of the organic ligand; otherwise there was no Hg(II) concentration variation at all.
Assuntos
Membranas Artificiais , Mercúrio/isolamento & purificação , Poluentes Químicos da Água/isolamento & purificação , Purificação da Água/instrumentação , Purificação da Água/métodos , Concentração de Íons de Hidrogênio , Mercúrio/análise , Mercúrio/química , Microscopia Eletrônica de Varredura , Ácido Pentético/química , Poluentes Químicos da Água/análiseRESUMO
The importance assigned to chelating agents in diverse areas has impelled studies concerning their development as related to metal ions representing a biological concern. The synthesis of di-isopropyliminodiacetoamide (D) is presented in this work. The acidity constant obtained for D was pKa = 5.79 +/- 0.04 with the aid of program SUPERQUAD. The equilibrium constants for D with Cu(II) were obtained with the aid of program SQUAD for CuD2+ and CuD2(2)+ species giving log beta1 = 4.795 +/- 0.002 and log beta2 = 8.374 +/- 0.004, respectively.