Glucose Biosensor Based on Oxygen Electrode Part III: Long-Term Performance of the Glucose Biosensor in Blood Plasma at Body Temperature

Abstract

A potentially implantable glucose biosensor for continuous monitoring of glucose levels in diabetic patients has been developed. The glucose biosensor is based on an amperometric oxygen electrode and Glucose Oxidase immobilized on carbon powder held in a form of a liquid suspension. The enzyme material can be replaced (the sensor recharged) without sensor disassembly. Glucose diffusion membranes from polycarbonate (PC) and from polytetrafluorethylene (PTFE) coated with silastic are used.

Sensors were evaluated continuously operating in phosphate buffer solution and in undiluted blood plasma at body temperature. Calibration curves of the sensors were periodically obtained. The sensors show stable performance during at least 1200 hours of operation without refilling of the enzyme. The PTFE membrane demonstrates high mechanical stability and is little effected by long-term operation in undiluted blood plasma.     

Glucose Biosensor Based on Oxygen Electrode. Part II: Long-Term Operational Stability of the Rechargeable Glucose Biosensor

Abstract

A potentially implantable glucose biosensor for measuring blood or tissue glucose levels in diabetic patients has been developed. The glucose biosensor is based on an amperometric oxygen electrode and immobilized glucose oxidase enzyme, in which the immobilized enzyme can be replaced (the sensor recharged) without surgical removal of the sensor from the patient. Recharging of the sensor is achieved by injecting fresh immobilized enzyme into the sensor using a septum. A special technique for immobilization of the enzyme on Ultra-Low Temperature Isotropic (ULTI) carbon powder held in a liquid suspension has been developed.

In vitro studies of the sensors show stable performance during several recharge cycles over a period of 3 months of continuous operation.

Diffusion membranes which ensure linear dependence of the sensor response on glucose concentration have been developed. These membranes comprise silastic latex-rubber coatings over a microporous polycarbonate membrane. Calibration curves of the amperometric signal show linearity over a wide range of glucose concentrations (up to 16 mM), covering hypoglycemic, normoglycemic and hyperglycemic conditions.

The experimental results confirm the suitability of the sensors for in vitro measurements in undiluted human sera.     

Glucose Biosensor Based on Oxygen Electrode Part IV: In Vivo Evaluation of the Rechargeable Glucose Sensor

Abstract

A glucose monitoring system consisting of a pair of amperometric sensors: a glucose biosensor based on oxygen electrode and an oxygen sensor, two miniature potentiostats, an instrumentation amplifier and a data logger has been developed. The glucose sensor has linear response to the glucose concentration in vitro at 37°C up to 26 mM (480 mg/dL) in the phosphate buffer solution (pH 7.4), and linear range up to 21 mM (380 mg/dL) in undiluted bovine plasma. The system was evaluated in vivo with the sensors subcutaneously implanted in healthy mongrel dogs. During the implantation the system output was continuously recorded. The results of short-term subcutaneous implantation of the integrated system demonstrated good agreement between the glucose concentration measured by the biosensor and that obtained using standard glucose determination methods. The delay-time between the tissue glucose level (measured by the biosensor) and the blood glucose level (obtained by standard methodology) was 3 to 10 minutes. During the chronic implantation the biosensor was refilled in vivo. Rejuvenation of the sensor response after refilling was observed demonstrating the potential of such sensors for long-term implantation.     

Measurement of Glucose Concentration in Blood Plasma Based on a Wireless Magnetoelastic Biosensor

Abstract

A wireless magnetoelastic glucose biosensor in blood plasma is described, based on using a mass sensitive magnetoelastic sensor as transducer. The glucose biosensor was fabricated by coating the ribbon‐like, magnetoelastic sensor with a pH sensitive polymer and a biolayer of glucose oxidase (GOx) and catalase. The pH response polymer swells or shrinks, thereby changing sensor mass loading, respectively, in response to increase or decrease of pH values. The GOx–catalyzed oxidation of the glucose in blood plasma produces gluconic acid, resulting in the pH sensitive polymer shrinking, which in turn decreases the sensor mass loading. The results show that the proposed magnetoelastic glucose biosensor can be successfully applied to determine the concentration of glucose in blood plasma. At glucose concentration range of 2.5–20.0 mmol/l, the biosensor responses are reversible and linear, with a detection limit of 1.2 mmol/l. Since no physical connections between the sensor and the monitoring instruments are required, this proposed biosensor can potentially be applied to in vivo and in situ measurement of glucose concentration in physiological fluids.     

Fluorescent Determination of Glucose Using Silicon Nanodots

ABSTRACT

Here is reported a fluorescent biosensor for glucose detection based on water-soluble and pH-responsive silicon nanodots. The silicon nanodots were prepared using a facile hydrothermal method. The advantages of using the silicon nanodots as glucose sensor are twofold. Firstly, the fluorescence of silicon nanodots was quenched by hydrogen peroxide that was produced from glucose oxidation. Secondly, the fluorescence of silicon nanodots was highly sensitive to gluconic acid that was also produced by glucose oxidation. Our results show that this method detected glucose as low as 0.54?µM with a good selectivity and allowed the determination of glucose in serum samples. This method is also simple, rapid, low-toxic and low-cost, thereby hold high application potential for biological assays.     

Hydrogel Membrane Improves Batch‐to‐Batch Reproducibility of an Enzymatic Glucose Biosensor

A permselective membrane is a critical component that defines the linear detection limits, the sensitivity, and thus the ultimate efficacy of an enzymatic biosensor. Although membranes like epoxy‐polyurethane (epoxy‐PU) and Nafion are widely used and provide the desired glucose detection limits of 2 to 30 mM, both the within batch and batch‐to‐batch variability of sensors that use these materials is a concern. The hypothesis for this study was that a crosslinked hydrogel would have a sufficiently uniform porosity and hydrophilicity to address the variability in sensor sensitivity. The hydrogel was prepared by crosslinking di‐hydroxyethyl methacrylate, hydroxyethyl methacrylate and N‐vinyl pyrrolidone with 2.5 mol% ethylene glycol dimethacrylate using water soluble initiators – ammonium persulfate and sodium metabisulfite under a nitrogen atmosphere. The hydrogel was applied to the sensor by dip coating during polymerisation. Electrochemical measurements revealed that the response characteristics of sensors coated with this membrane are highly consistent. Scanning electrochemical microscopy (SECM) was used to spatially resolve glucose diffusion through the membrane by measuring the consequent H₂O₂ release and compared with an epoxy‐PU membrane. Hydrogen peroxide measurements using SECM revealed that the epoxy‐PU membranes had uneven lateral diffusion profiles compared to the uniform profile of the hydrogel membranes. The uneven diffusion profiles of epoxy‐PU membranes are attributed to a fabrication method that results in uneven membrane properties, while the uniform diffusion profiles of the hydrogel membranes are primarily dictated by their uniform pore size.     

Glucose Biosensor Based on Oxygen Electrode. Part I: Silastic Coated Polycarbonate Membranes for Biosensor Application

Abstract

An amperometric glucose biosensor based on the oxygen electrode principle has been developed. Polycarbonate membranes (pore size from 0.01 μm to 0.4 μm) were used as external glucose diffusion membranes in order to obtain direct proportionality of the amperometric signal to the substrate concentration in the entire physiological range. The commercially available membranes - standard (hydrophilic, treated with Polyvinylpyrrolidone/(PVP)) and PVP-free membranes were compared with membranes coated with a silicone elastomer (silastic). Spindrop coating technique was used to create stable, adhesive coatings over the polycarbonate membranes. These coated membranes achieved diffusion control of the glucose flux such that the amperometric signal of the biosensor was linearly proportional to the substrate concentration up to 16 mM glucose. The membrane parameters were optimized by varying the parameters of the coating process-spin rate of the membrane rotation and the silastic/water ratio in the coating emulsion.     

Amperometric Glucose Biosensor Based on Ultrafine Platinum Nanoparticles

Abstract

In the present paper the ultrafine and highly dispersed platinum nanoparticles (average size 3 nm) were used for the construction of a glucose biosensor in a simple method. An excellent response to glucose has been obtained with a high sensitivity (137.7 µA mM^?1 cm^?2) and fast response time (5 s). The biosensor showed a detection limit of 5 µM (at the ratio of signal to noise, S/N=3) and a linear range form 0.2 to 3.2 mM with a correlation coefficient r=0.999. The apparent Michaelis–Menten constant (k _m) and the maximum current were estimated to be 9.36 and 1.507 mA mM^?1 cm^?2, respectively. In addition, effects of pH value, applied potential and the interferents on the amperometric response of the sensor were investigated and discussed.     

A Novel Potentiometric Glucose Biosensor Based on Polyaniline Semiconductor Films

Abstract

Application of polyaniline semiconductor films to potentiometric biosensor development provides certain advantages comparing with the known systems. Using self-doped polyaniline instead of common polymer as pH transducer the stable potentiometric response of 70 mV/pH was obtained. Taking as an example glucose biosensor we showed that polyaniline based electrode possessed three-four fold increased potential shift than glucose-sensitive field-effect transistor did. One can increase the sensitivity of potentiometric biosensor using thick ion-exchange membranes (in our case Nafion) in order to concentrate product near electrode surface. Such sensor possessed higher response time.     

Amperometric Glucose Sensor Fabricated by Combining Glucose Oxidase Micelle Membrane and Aminated Glassy Carbon Electrode

Abstract

An amperometric glucose biosensor based on the detection of the reduction of oxygen has been developed by combining an aminated glassy carbon electrode with a polystyrene (PS) membrane containing glucose oxidase (GOD) micelles. The structure of GOD micelles contained in PS membrane was observed by scanning electron microscope. The micelle has a roughly spherical shape, and the enzyme colony is contained inside the micelle. This glucose sensor exhibited good sensitivity with short response time (within 2 min). A good linear relationship was observed in the concentration range of 0.2 mM to 2.6 mM when the applied potential was ? 0.45 V vs. Ag/AgCl.     

The Nitrite Oxidizing Activity of Nitrobacter Strains as a Base of Microbial Biosensor for Nitrite Detection

ABSTRACT

Parameters of three Nitrobacter based sensor models have been studied for the purpose of creation of a nitrite analyzer that could be used for assay of nitroaromatics. It was shown that the sensor based on N. vulgaris strain was most suitable for development of such a device. The sensor possessed high     

Biosensors for determination of glucose with glucose oxidase immobilized on an eggshell membrane

A glucose biosensor using an enzyme-immobilized eggshell membrane and oxygen electrode for glucose determination has been fabricated. Glucose oxidase was covalently immobilized on an eggshell membrane with glutaraldehyde as a cross-linking agent. The glucose biosensor was fabricated by positioning the enzyme-immobilized eggshell membrane on the surface of a dissolved oxygen sensor. The detection scheme was based on the depletion of dissolved oxygen content upon exposure to glucose solution and the decrease in the oxygen level was monitored and related to the glucose concentration. The effect of glutaraldehyde concentration, pH, phosphate buffer concentration and temperature on the response of the glucose biosensor has been studied in detail. Common matrix interferents such as ethanol, d-fructose, citric acid, sodium benzoate, sucrose and l-ascorbic acid did not give significant interference. The resulting sensor exhibited a fast response (100 s), high sensitivity (8.3409 mg L⁻¹ oxygen depletion/mmol L⁻¹ glucose) and good storage stability (85.2% of its initial sensitivity after 4 months). The linear response is 1.0×10⁻⁵ to 1.3×10⁻³ mol L⁻¹ glucose. The glucose content in real samples such as commercial glucose injection preparations and wines was determined, and the results were comparable to the values obtained from a commercial glucose assay kit based on a spectrophotometric method.     

Acid Phosphatase Assay with a Wireless Magnetoelastic Biosensor

Abstract

A wireless remote‐query disposable magnetoelastic (ME) biosensor was developed for the assay of acid phosphatase (ACP). The sensor was fabricated by applying a magnetoelastic ribbon with a layer of pH‐sensitive polymer and upon it a sensing film containing bovine serum albumin (BSA) and adenosine‐5′‐monophosphate (5′‐AMP). The ACP‐catalyzed hydrolysis of 5′‐AMP decreases the solution pH, resulting in the polymer shrinking and consequently the resonance frequency of the magnetoelastic sensor increasing. The kinetic parameters were measured to be 1.64×10^?3 M (Michaelis constant) and 130 Hz/min (maximum initial rate). The proposed sensor can determine 0.2 to 1.2 U/ml of ACP.     

A Novel Microbial Sensor for Anionic Surfactant Determination

Abstract

A novel whole cell biosensor was constructed for detection of anionic surfactants in an aquatic environment. The sensor response to linear alkyl benzene sulfonates was linear up to 6 mg l^?1 which is a range suitable for the detection of anionic surfactant concentration in polluted river water. Under optimum conditions, the sensor response time was less than 15 min. Anionic surfactant analysis was rapid and convenient and did not require organic reagents which are harmful to an environment.     

Optical Fiber Chemiluminescence Biosensor for Antioxidants Based on an Immobilized Luminol/Hematin Reagent Phase

ABSTRACT

A chemiluminescence (CL) biosensing system for antioxidants was developed based on luminol and hematin co-immobilized on a cellulose membrane disc. The concentration of the antioxidant was quantified through the measurement of the inhibition of the CL emitted when hydrogen peroxide was introduced into the reagent phase. The instrumentation employed in the measurement was a fabricated luminometer employing optical fibers and a UV-enhanced photodiode transducer. Under optimum conditions, linear calibration curves were obtained for antioxidant concentrations ranging from 1 × 10^-4 M to 0.10 M, with an average relative standard deviation of about 5%. The minimum detectable concentration was 100 μM, and the response time was less than 60 seconds. The sensor response correlated well (r = 0.9979) with the results of a standard colorimetric method for a specific antioxidant (propyl gallate). The sensor was used to assess the antioxidant capacity of water infusions prepared from the dried leaves of some Philippine medicinal plants.     

Acid Phosphatase Assay with a Wireless Magnetoelastic Biosensor

Abstract

A wireless remote‐query disposable magnetoelastic biosensor is developed for the assay of acid phosphatase (ACP). The sensor was fabricated by applying a layer of pH‐sensitive polymer to a magnetoelastic ribbon and, on top of it, a sensing film containing bovine serum albumin and adenosine‐5′‐monophosphate (5′‐AMP), the substrate of ACP. In response to an externally applied time‐varying magnetic field, the magnetoelastic sensor mechanically vibrates at a characteristic frequency that is inversely dependent on the mass of the attached film. Because the magnetoelastic sensor is magnetostrictive, the mechanical vibrations of the sensor launch magnetic flux that can be detected remotely from the sensor using a pick‐up coil. The ACP‐catalyzed hydrolysis of 5′‐AMP decreases the solution pH, resulting in the polymer shrinking and, consequently, the resonance frequency of the magnetoelastic sensor increasing. The experimental condition was optimized to be 37°C, in 2.5 mM sodium citrate buffer solution (pH 6.5). Using citrate as buffer can enhance the ACP activity, partly offsetting the effect of the buffer capacity on the sensitivity. The kinetic parameter were measured to be: 1.64×10^–3 M (Michaelis constant) and 130 Hz/min (maximum initial rate). This work provides a remote enzymatic assay of ACP. The relative standard deviation in the measurements of six sensors in parallel is 3.4%. The proposed sensor can determine 0.2～1.2 U/ml of ACP.     

Activation‐Based Catalase Enzyme Electrode and its Usage for Glucose Determination

Abstract

In this study, a novel type amperometric biosensor, which is based on the activation of catalase enzyme by glucose, was developed and used for the sensitive determination of glucose. For the preparation of the biosensor catalase enzyme was immobilized in gelatin by using cross‐linking agent glutaraldehyde and fixed on a pretreated teflon membrane of a dissolved oxygen probe. Glucose was used as an activator for the catalase enzyme and determination of glucose is based on the assay of the differences on the catalase activity of the biosensor on the oxygenmeter in the absence and the presence of glucose in the reaction medium. The responses of the activation based catalase biosensor have a linear relation to glucose concentrations and good measurement correlation between 0.5 and 5.0 µM with 2 min response time. In the optimization studies of the biosensor the most suitable catalase amount were found as 1324 U cm^?2 and also phosphate buffer (pH: 7.0; 50 mM) and 35°C were obtained as the optimum working conditions. For the characterization studies of the biosensor some parameters such as activator and interference effects of some substances on the biosensor response, reproducibility and operational stability were performed.     

基于二氧化钛/碳纳米管/壳聚糖纳米复合薄膜制备葡萄糖生物传感器

利用壳聚糖(CHI)溶液分散了纳米二氧化钛(nano-TiO2)和多壁碳纳米管(MWCNT),将该分散液修饰于玻碳电极表面形成纳米复合薄膜;用戊二醛为交联剂在该纳米复合层上固定了葡萄糖氧化酶(GOx),同时以二茂铁为电子媒介体构建了一种新型葡萄糖传感器。利用扫描电镜(SEM)、交流阻抗(AC)对所制备的传感器进行了表征,同时用循环伏安法(CV)和计时电流法(CA)考察了其对葡萄糖的电催化氧化性能。实验结果表明,在优化测试条件下该传感器对葡萄糖在0.5～20.0 mmol.L-1范围内有线性响应,检出限为0.2 mmol.L-1;电流达到95%的稳态时间小于5 s;此生物传感器具有良好的重现性和选择性,能有效排除抗坏血酸、尿酸等常见干扰物的影响并成功应用于饮料中葡萄糖含量的测定。     

Effect of Urea on the Morphology of Co3O4 Nanostructures and Their Application for Potentiometric Glucose Biosensor

In this study, an effect of different concentrations of urea on the morphology of cobalt oxide (Co₃O₄) nanostructures was investigated. The Co₃O₄ nanostructures are fabricated on gold coated glass substrate by the hydrothermal method. The morphological and structural characterization was performed by scanning electron microscopy, and X‐ray diffraction techniques. The Co₃O₄ nanostructures exhibit morphology of flowers‐like and have comprised on nanowires due to the increasing amount of urea. The nanostructures were highly dense on the substrate and possess a good crystalline quality. The Co₃O₄ nanostructures were successfully used for the development of a sensitive glucose biosensor. The presented glucose biosensor detected a wide range of glucose concentrations from 1×10^?6 M to 1×10^?2 M with sensitivity of a ?56.85 mV/decade and indicated a fast response time of less than 10 s. This performance could be attributed to the heterogeneous catalysis effect at glucose oxidase enzyme, nanoflowers, and nanowires interfaces, which have enhanced the electron transfer process on the electrode surface. Moreover, the reproducibility, repeatability, stability and selectivity were also investigated. All the obtained results indicate the potential use of the developed glucose sensor for monitoring of glucose concentrations at drugs, human serum and food industry related samples.     

Glucose Oxidase Electrode Based on Graphite and Methylthio Tetrathiafulvalene as Mediator

Abstract

A glucose sensor based on glucose oxidase and a new mediator - 4,5-dimethyl-4′-methylthio-Δ 2,2′-bi-1,3-dithiole (MTTTF) is described. The background for sensor action is the effective MTTTF cation interaction (apparent bimolecular constant (2.0+/-0.5)?10⁶ M^?1 s^?1 at 25°C and pH 7.0) with reduced glucose oxidase and the high electrochemical rate of mediator transformation.

A glucose sensor was prepared by adsorbing mediator (MTTTF) and glucose oxidase on graphite rods. The sensor responds to glucose at electrode potentials higher than 50 mV vs SCE, but the maximal activity is obtained at a potential of 250 mV. In air saturated solution the electrode shows a non-linear calibration curve with a half-saturation concentration 10.4 mM and Hill coefficient 2.08 at 250 mV. Sensor response changes little at pH 6.5–8.0. The energy of activation of the sensor response calculated from the Arrhenius equation was 64.5 kJ/mol, and the temperature coefficient at 25°C was 9.2%.