Amperometric uric acid sensors based on polyelectrolyte multilayer films

Uricase (UOx) and polyelectrolyte were used for preparation of a permselective multilayer film and enzyme multilayer films on a platinum (Pt) electrode, allowing the detection of uric acid amperometrically. The polyelectrolyte multilayer (PEM) film composed of poly(allylamine) (PAA) and poly(vinyl sulfate) (PVS) were prepared via layer-by-layer assembly on the electrode, functioning as H₂O₂-selective film. After deposition of the permselective film (PAA/PVS)₂PAA, UOx and PAA were deposited via layer-by-layer sequential deposition up to 10 UOx layers to prepare amperometric sensors for uric acid. Current response to uric acid was recorded at +0.6 V vs. Ag/AgCl to detect H₂O₂ produced from the enzyme reaction. The response current increased with increasing the number of UOx layers. Even in the presence of ascorbic acid, uric acid can be detected over the concentration range 10⁻⁶-10⁻³ M. The response current and deposited amount of UOx were affected by deposition bath pH and the addition of salt. The deposition of PAA/UOx film prepared in 2 mg ml⁻¹ solution (pH 11) of PAA with NaCl (8 mg ml⁻¹) and 0.1 mg ml⁻¹ solution (pH 8.5) of UOx with borate (100 mM) resulted in an electrode which shows the largest response to uric acid. The response of the sensor to uric acid was decreased by 40% from the original activity after 30 days.     

Assay for uric acid level in rat striatum by a reagentless biosensor based on functionalized multi-wall carbon nanotubes with tin oxide

A novel reagentless amperometric uric acid biosensor based on functionalized multi-wall carbon nanotubes (MWCNTs) with tin oxide (SnO₂) nanoparticles has been developed. This was successfully applied to assay uric acid levels from an in vivo microdialysis sampling. Compared with unfunctionalized or traditional carboxylic acid (–COOH)-functionalized MWCNTs, the MWCNTs–SnO₂ electrode exhibited higher electrocatalytic oxidation to uric acid. Here, MWCNTs–SnO₂ may act as an efficient promoter, and the system exhibited a linear dependence on the uric acid concentration over the range from 1.0 × 10^–7 to 5.0 × 10^–4 mol L^–1. In addition, there was little ascorbic acid interference. The high sensitivity of the MWCNTs–SnO₂ modified enzyme electrode enabled the monitoring of trace levels of uric acid in dialysate samples in rat striatum.     

邻菲啰啉对固定尿酸酶催化反应动力学的影响

The activation energy of the enzyme-catalyzed reaction for uric acid decreases markedly in the presence of o-phenanthroline, which activates the bioelectrochemicla activity of the polypyrrole uricase electrode. The response current of the enzyme electrodeis independent of the concentration of o-phenanthroline. Based on the experimental results, the mechamsm of the enzyme-catalyzed reaction for uric acid in the presence of o-phenanthroline is presented as follows: E+A→EA, EA+S EAS, EAS→EA+P, where E, A, S and P are the enzyme, activator, substrate and product, respectively. The effects of pH value, potential and the uric acid concentration on the response currents of the uricase electrode have been studied in the presence of o-phenanthroline. In the presence of o-phenanthroline, the response current of the enzyme electrode increase linearly with increasing concentration of uric acid in the region of 0.07 to 0.67 mmol·L~(-1), therefore the polypyrrole uricase electrode which has once lost its activity can be activated and used again to determine the substrate concentration.     

Amperometric determination of ascorbic acid on screen-printing ruthenium dioxide electrode

Ruthenium oxide (RuO₂) is commonly used in resistive pastes for screen printing. The electrochemical properties of a screen-printing planar RuO₂ electrode have hardly been studied. In this communication, planar electrochemical sensors with a RuO₂ working electrode, an Ag|AgCl reference electrode and a RuO₂ counter electrode are fabricated by screen printing. The electro-oxidation of ascorbic acid, uric acid, and hydrogen peroxide on these RuO₂ electrodes is investigated by means of cyclic voltammetry. Compared to uric acid and hydrogen peroxide, ascorbic acid can be easily oxidized at the low operating potential (<150 mV versus Ag|AgCl). The amperometric measurement of ascorbic acid at 100 mV on a RuO₂ electrode shows fast response and good linearity in the 0–4 mM range. Meanwhile, the electrochemical interference from uric acid and hydrogen peroxide at this potential is very small.     

Fabrication of dissolved O2 metric uric acid biosensor using uricase epoxy resin biocomposite membrane

Uricase purified from 20-day-old leaves of cowpea was immobilized on to epoxy resin membrane with 80% retention of initial activity of free enzyme and a conjugation yield of 0.056 mg/cm². The uricase epoxy resin bioconjugate membrane was mounted over the sensing part of the combined electrode of ‘Aqualytic’ dissolved O₂ (DO) meter to construct a uric acid biosensor. The biosensor measures the depletion of dissolved O₂ during the oxidation of uric acid by immobilized uricase, which is directly proportional to uric acid concentration. The biosensor showed optimum response within 10-12 s at a pH 8.5 and 35 °C. A linear relationship was found between uric acid concentration from 0.025 to 0.1 mM and O₂ (mg/l) consumed. The biosensor was employed for measurement of uric acid in serum. The mean value of uric acid in serum was 4.92 mg/dl in apparently healthy males and 3.11 mg/dl in apparently healthy females. The mean analytic recoveries of added uric acid in reaction mixture (8.9 and 9.8 mg/dl) were 93.6 ± 2.34 and 87.18 ± 3.17% respectively. The within and between batch CVs were <6.5 and <5.0%, respectively. The serum uric acid values obtained by present method and standard enzymic colorimetric method, showed a good correlation (r = 0.996) and regression equation being y = 0.984x + 0.0674. Among the various metabolites tested only, glucose (11%), urea (38%), NaCl (25%) and cholesterol (13%) and ascorbic acid (56%) caused decrease, while, MgSO₄ and CaCl₂ had no effect on immobilized enzyme. The enzyme electrode showed only 32% decrease during its use for 100 times over a period of 60 days at 4 °C.     

Carbon Paste Electrode Modified with ZrO<Subscript>2</Subscript> Nanoparticles and Ionic Liquid for Sensing of Dopamine in the Presence of Uric Acid

A novel carbon paste electrode modified with ZrO₂ nanoparticles and an ionic liquid (n-hexyl-3- methylimidazolium hexafluorophosphate) was fabricated. The electrochemical study of the modified electrode, as well as its efficiency for simultaneous voltammetric oxidation of dopamine and uric acid is described. The electrode was also employed to study the electrochemical oxidation of dopamine and uric acid, using cyclic voltammetry, chronoamperometry and square wave voltammetry as diagnostic techniques. Square wave voltammetry exhibits linear dynamic range from 1.0 × 10^?6 to 9.0 × 10^?4 M for dopamine. Also, square wave voltammetry exhibits linear dynamic range from 9.0 × 10^?6–1.0 × 10^?3 M for uric acid. The modified electrode displayed strong function for resolving the overlapping voltammetric responses of dopamine and uric acid into two well-defined voltammetric peaks. In the mixture containing dopamine and uric acid, the two compounds can be well separated from each other with potential difference of 155 mV, which is large enough to determine dopamine and uric acid individually and simultaneously. Finally, the modified electrode was used for determination of dopamine and uric acid in real samples.     

Preparation and application of a new glucose sensor based on iodide ion selective electrode

An electrode for glucose has been prepared by using an iodide selective electrode with the glucose oxidase enzyme. The iodide selective electrode used was prepared from 10% TDMAI and PVC according our previous study. The enzyme was immobilized on the iodide electrode by holding it at pH 7 phosphate buffer for 10 min at room temperature. The H₂O₂ formed from the reaction of glucose was determined from the decrease of iodide concentration that was present in the reaction cell. The iodide concentration was followed from the change of potential of iodide selective electrode. The potential change was linear in the 4×10⁻⁴ to 4×10⁻³ M glucose concentration (75-650 mg glucose/100ml blood) range. The slope of the linear portion was about 79 mV per decade change in glucose concentration. Glucose contents of some blood samples were determined with the new electrode and consistency was obtained with a colorimetric method. The effects of pH, iodide concentration, the amount of enzyme immobilized and the operating temperature were studied. No interference of ascorbic acid, uric acid, iron(III) and Cu(II) was observed. Since the iodide electrode used was not an AgI-Ag₂S electrode, there was no interference of common ions such as chloride present in biological fluids. The slope of the electrode did not change for about 65 days when used 3 times a day.     

An Electrochemical Determination of Uric Acid in Sera by a Flow-Through Equipment with and Without Uricase

Abstract

A comparison of an enzymeless direct electrochemical oxidation procedure at a platinum electrode for the determination of uric acid, and an enzyme sensor with immobilized urate: oxygen oxidoreductase (uricase), was performed in flow stream systems. The uricase enzyme electrode is based on the H₂O₂ oxidation current. Both amperometric methods were related to the wall-known photometric uricase-catalase-procedure (UCM) as a reference method. The measured values of both methods are of the first derivatives of current change (dI/dt) due to the electrochemical or electrochemical enzymatic reaction, respectively. The analytical quality of the measurements is characterized by: precision s% within run < 2% day to day < 5% accuracy acceptable (control materials) correlation to reference method r >0.93 analysis rate 80 samples/hr     

Electrochemical Enhanced Detection of Uric Acid Based on Peroxidase-like Activity of Fe3O4@Au

Fe₃O₄@Au nanomaterials were prepared by the self-assembly method. An enzyme-free, ultrasensitive electrochemical detection of uric acid was achieved based on the peroxidase-like activity of Fe₃O₄@Au. The proposed procedure has exhibited excellent catalytic activities and achieved significant enhancements of the current responses to uric acid. The detection range was from 0.1 to 10 mmol/L, and the limit of detection was 0.087 μmol/L. Under the action of external magnetic field, the magnetic particles can be easily separated from the bottom liquid, which has the advantages of simple operation and high separation efficiency. Moreover, this detection method combining a simulated enzyme and electrochemical can enhance the effective output of the overall electrochemical signal without modifying the electrode, and excellent reproducibility can be achieved. Compared to colorimetric assay, the electrochemical one has higher sensitivity and selectivity, and was further applied in ultrasensitive detection of uric acid in food samples. In short, the proposed electrochemical assay has great potential in the fields of food quality control and biomedical analysis.     

Electrochemical Behaviors of Ascorbic Acid and Uric Acid in Ionic Liquid

Cyclic voltammetric measurements at platinum electrode have been carried out to investigate the electrochemical oxidation of ascorbic acid and uric acid in ionic liquid, [bmim][BF₄]. It is important that a typical redox couple of ascorbic acid was obtained and it is oxidized to dehydroascorbic acid in [bmim][BF₄]. However, there is no electron-transfer for uric acid and no electrochemical oxidation carried out in the same ionic liquid. It provides a new way to eliminate the interfering between ascorbic acid and uric acid in the study of the electrochemical behaviors for them.     

Amperometric biosensor based on thermally activated polymer-stabilized metal nanoparticles

Polymer-stabilized Pd nanoparticles on carbon support were synthesized by a low thermal procedure that does not involve the utilization of a reducing agent such as NaBH₄ or hydrogen gas for the formation of the metallic nanoparticles. The Pd-catalyzed graphite particles were then mixed with known amounts of glucose oxidase (GOx) enzyme and Nafion to prepare a GOx-immobilized ink. A glassy carbon electrode (GCE) modified with the GOx ink was used to evaluate the performance of the biosensor electrode. The results of TEM and AFM show that the Pd nanoparticles are uniformly distributed on top of the substrate. Results are presented for sensing glucose through the voltammetric measurement of H₂O₂. Coupled with the simplicity of preparation, the biosensor exhibited high sensitivity and extended linear range for glucose measurement. Further, the electrochemical characteristics of the nanocomposite biosensor were evaluated with respect to the electrochemistry of potassium ferricyanide by cyclic voltammetry. Whereas the presence of polymer and Nafion improved the stability of both the ink and biosensor electrode, the concentration of glucose was measured without interferences from oxygen, ascorbic acid and uric acid because of the Nafion.     

A highly selective and sensitive dopamine and uric acid biosensor fabricated with functionalized ordered mesoporous carbon and hydrophobic ionic liquid

An ordered mesoporous carbon material functionalized with carboxylic acid groups was synthesized. It was characterized by powder X-ray diffraction, transmission electron microscopy, Fourier transform IR spectroscopy and N₂ adsorption/desorption. Furthermore, this material was used to modify an electrode surface combined with a hydrophobic ionic liquid. The functionalized ordered mesoporous carbon/ionic liquid gel modified electrode shows excellent electrocatalytic performances for the oxidation of dopamine, uric acid and ascorbic acid. The presence of the ionic liquid promotes the electron transfer. Linear responses for dopamine and uric acid were obtained in the ranges of 0.1 to 500 μM and from 0.1 to 100 μM with detection limits of 4.1 and 2.5 nM (signal-to-noise ratio of 3), respectively, under optimum conditions. A quick and sensitive biosensor based on functionalized ordered mesoporous carbon and an ionic liquid has been developed for the first time for the detection of dopamine and uric acid in the presence of a large amount of ascorbic acid.     

Voltammetric Determination of Uric Acid at a Fullerene‐C60‐Modified Glassy Carbon Electrode

Glassy carbon electrode modified by microcrystals of fullerene‐C₆₀ mediates the voltammetric determination of uric acid (UA) in the presence of ascorbic acid (AA). Interference of AA was overcome owing to the ability of pretreated fullerene‐C₆₀‐modified glassy carbon electrode. Based on its strong catalytic function towards the oxidation of UA and AA, the overlapping voltammetric response of uric acid and ascorbic acid is resolved into two well‐defined voltammetric peaks with lowered oxidation potential and enhanced oxidation currents under conditions of both linear sweep voltammetry (LSV) and Osteryoung square‐wave voltammetry (OSWV). At pH 7.2, a linear calibration graph is obtained for UA in linear sweep voltammetry over the range from 0.5 μM to 700 μM with a correlation coefficient of 0.9904 and a sensitivity of 0.0215 μA μM^?1 . The detection limit (3σ) is 0.2 μM for standard solution. AA in less than four fold excess does not interfere. The sensitivity and detection limit in OSWV were found as 0.0255 μA μM^?1 and 0.12 μM, for standard solution respectively. The presence of physiologically common interferents (i.e. adenine, hypoxanthine and xanthine) negligibly affects the response of UA. The fullerene‐C₆₀‐modified electrode exhibited a stable, selective and sensitive response to uric acid in the presence of interferents.     

Electrochemical determination of uric acid using a mesoporous SiO2-modified electrode

A mesoporous SiO₂-modified carbon paste electrode for the determination of uric acid is described. Owing to the regular and specific mesoporous channels, numerous active sites and a large surface area, the mesoporous SiO₂-modified electrode greatly increases the oxidation peak current of uric acid. Based on this, a highly sensitive, rapid and convenient electrochemical method was developed for the determination of uric acid after optimizing the experimental parameters (supporting electrolyte, content of mesoporous SiO₂, accumulation potential and time). The linear range is from 2.5 × 10⁻⁷ to 2.0 × 10⁻⁵ mol L⁻¹, and the limit of detection is estimated to be 8.0 × 10⁻⁸ mol L⁻¹. The relative standard deviation for 10 mesoporous SiO₂-modified electrodes is 5.8%. The method was used to determine uric acid in human serum samples. Correspondence: Kangbing Wu, Department of Chemistry, Huazhong University of Science and Technology, Wuhan 430074, P.R. China     

Simultaneous Sensitive Determination of Dopamine and Uric Acid in the Presence of Excess Ascorbic Acid with a Magnetic Chitosan Microsphere/Thionine Modified Electrode

Magnetic chitosan microspheres (MCMS) and thionine were incorporated in a modified electrode for the simultaneous sensitive determination of dopamine (DA) and uric acid (UA). Due to the unique properties of the MCMS and the electron mediation of thionine, this modified electrode showed excellent electrocatalytic oxidation toward dopamine and uric acid with a large separation of peak potentials and a significant enhancement of peak currents. However, the electrochemical behavior of ascorbic acid may be depressed at the modified electrode. Differential pulse voltammetry was used for the simultaneous sensitive determination of dopamine and uric acid in the presence of excess ascorbic acid at this modified electrode. The current responses showed excellent linear relationships in the range of 2–30 µM and 9–100 µM for dopamine and uric acid, respectively. The detection limits were estimated to be 0.5 µM and 2.3 µM for dopamine and uric acid, respectively. In addition, this modified electrode showed excellent repeatability, good stability, and satisfactory reliability, thus indicating potential for the practical applications.     

Immobilized Fullerene C60‐Enzyme‐Based Electrochemical Glucose Sensor

A mixed‐valence cluster of cobalt(II) hexacyanoferrate and fullerene C60‐enzyme‐based electrochemical glucose sensor was developed. A water insoluble fullerene C60‐glucose oxidase (C60‐GOD) was prepared and applied as an immobilized enzyme on a glassy carbon electrode with cobalt(II) hexacyanoferrate for analysis of glucose. The glucose in 0.1 M KCl/phosphate buffer solution at pH = 6 was measured with an applied electrode potential at 0.0 mV (vs Ag/AgCl reference electrode). The C60‐GOD‐based electrochemical glucose sensor exhibited efficient electro‐catalytic activity toward the liberated hydrogen peroxide and allowed cathodic detection of glucose. The C60‐GOD electrochemical glucose sensor also showed quite good selectivity to glucose with no interference from easily oxidizable biospecies, e.g. uric acid, ascorbic acid, cysteine, tyrosine, acetaminophen and galactose. The current of H₂O₂ reduced by cobalt(II) hexacyanoferrate was found to be proportional to the concentration of glucose in aqueous solutions. The immobilized C60‐GOD enzyme‐based glucose sensor exhibited a good linear response up to 8 mM glucose with a sensitivity of 5.60 × 10² nA/mM and a quite short response time of 5 sec. The C60‐GOD‐based glucose sensor also showed a good sensitivity with a detection limit of 1.6 × 10^‐6 M and a high reproducibility with a relative standard deviation (RSD) of 4.26%. Effects of pH and temperature on the responses of the immobilized C60‐GOD/cobalt(II) hexacyanoferrate‐based electrochemical glucose sensor were also studied and discussed.     

A novel approach for the determination of paracetamol based on the reduction of N-acetyl-p-benzoquinoneimine formed on the electrochemically treated pencil graphite electrode

A novel approach was proposed for the simple and rapid electrochemical determination of paracetamol (PC) in the presence of uric acid in body fluids. The voltammetric determination of PC is based on the electrochemical reduction of N-acetyl-p-benzoquinoneimine formed simultaneously on the electrochemically treated pencil graphite (ETPG) electrode during the measurement. ETPG electrodes were prepared by the potential cycling between −0.3 V and +2.0 V in 0.1 M H₃PO₄ solution. The electrochemical performance of the ETPG electrode was evaluated by adsorptive transfer stripping differential pulse voltammetry (ATSDPV). The resulting sensor showed good performance for the determination of PC in human blood serum samples with a linear range of 0.05–2.5 μM and a highly reproducible response (RSD of 3.1%). The calculated detection limit was 2.5 nM (S/N = 3). The proposed method does not require any sample pretreatment, prevents the interference of uric acid and allows the determination of PC directly in blood serum samples.     

Eliminating the Interference of Ascorbic Acid and Uric Acid to the Amperometric Glucose Biosensor by Cation Exchangers Membrane and Size Exclusion Membrane

The characteristics of a multiuse planar amperometric biosensor modified with Nafion and/or polyion membrane were investigated. A new enzyme immobilization process was proposed, in which the polyvinyl alcohol bearing a styrylpyridinium (SbQ)/glucose oxidase composite was treated with glutaraldehyde vapor prior to the photocrosslinking reaction. The resulting planar enzyme electrode remains active for at least 150 days. Compared with poly‐L ‐lysine/poly (4‐stryenesulfonate) polyion complex membrane the Nafion membrane showed marked effect to reduce the electrochemical response of the modified planar enzyme electrode to the biological interferents, such as ascorbic acid and uric acid. Furthermore, Nafion membrane can effective restricting the oxidized anionic interferent to adhear on its surface, thereby the fouling of the electrode was avoided.     

Preparation of MoS2‐graphene Hybrid Nanosheets and Simultaneously Electrochemical Determination of Levodopa and Uric Acid

MoS₂ nanoflakes were prepared by exfoliating commercial MoS₂ powders with the assistance of ultrasound and graphene foam was synthesized by chemical vapor deposition using nickel foam as the template. MoS₂‐graphene hybrid nanosheets were developed through the combination of MoS₂ nanoflakes and graphene nanosheets by ultrasonic dispersion. The hybrid nanosheets were sprayed onto the ITO coated glass, which acts as an electrode for the simultaneously electrochemical determination of levodopa and uric acid. The MoS₂‐graphene hybrid nanosheets were characterized by scanning electron microscopy, X‐ray diffraction and Raman spectroscopy. The results show that the hybrid nanosheets are composed of MoS₂ and graphene with a sheet‐like morphology. The sensitivity of the electrode for levodopa and uric acid is 0.36 μA μM⁻¹ and 0.39 μA μM⁻¹, respectively. The electrode also shows low limit of detection, good selectivity, reproducibility and stability. And it is potential for use in clinical research.     

Gold nanoparticles on a thiol-functionalized silica network for ascorbic acid electrochemical detection in presence of dopamine and uric acid

A simple preparation methodology able to stabilize gold nanoparticles and to obtain an electrode which detects ascorbic acid, uric acid, and dopamine by different techniques is presented. A 3-mercaptopropyl-functionalized silica network was synthesized using the sol–gel method. Gold nanoparticles (nAu) were immobilized on the material at synthesis by adding a sol of these previously prepared particles to the reaction mixture. The electrochemical behavior of the SiO₂/MPTS/Au carbon paste electrode was studied using cyclic voltammetry in the presence of a hexacyanoferrate probe molecule. The presence of nAu in the functionalized silica network changes the electrochemical characteristics of the material, favoring the electron transfer process of this complex ion. The SiO₂/MPTS/Au electrode was proven to be an efficient tool in the simultaneous determination of ascorbic acid (H₂AA), dopamine (DA), and uric acid (UA) using square wave voltammetry techniques. With the nAu on the electrode, an increase in the peak current related to the redox process of the H₂AA, DA, and UA was observed. The separations of the anodic peak potentials between DA/H₂AA and UA/H₂AA were 310 and 442?mV, respectively. The results obtained show that the SiO₂/MPTS/Au electrode can be used in the simultaneous determination of H₂AA, DA, and UA.