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.     

Nitric Oxide Release for Enhanced Biocompatibility and Analytical Performance of Implantable Electrochemical Sensors

The real-time, continuous monitoring of glucose/lactate, blood gases and electrolytes by implantable electrochemical sensors holds significant value for critically ill and diabetic patients. However, the wide-spread use of such devices has been seriously hampered by implant-initiated host responses (e. g., thrombus formation, inflammatory responses and bacterial infection) when sensors are implanted in blood or tissue. As a result, the accuracy and usable lifetime of in vivo sensors are often compromised. Nitric oxide (NO) is an endogenous gas molecule able to inhibit platelet adhesion/activation, inflammatory responses and bacterial growth. As such, the release of NO from the surfaces of in vivo sensors is a promising strategy for enhancement of their biocompatibility and analytical performance. In this review, the physiological functions of NO to improve the biocompatibility of implantable electrochemical sensors are introduced, followed by a brief analysis of chemical approaches to realize NO release from such devices. A detailed summary of the various types of NO releasing electrochemical sensors reported to date and their performance in benchtop and/or in vivo testing are also provided. Finally, the prospects of future developments to further advance NO releasing sensor technology for clinical use are discussed.     

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.     

Improved thromboresistance and analytical performance of intravascular amperometric glucose sensors using optimized nitric oxide release coatings

In this work, nitric oxide (NO) release coatings designed for intravenous amperometric glucose sensors are optimized through the use of a polylactic acid (PLA) layer doped with a lipophilic diazeniumdiolated species that releases NO through a proton-driven mechanism. An Elast-Eon E2As polyurethane coating is used to both moderate NO release from the sensor surface and increase the sensor''s linear detection range toward glucose. These sensors were evaluated for thromboresistance and in vivo glucose performance through implantation in rabbit veins. By maintaining NO flux on a similar scale to endogenous endothelial cells, implanted glucose sensors exhibited reduced surface clot formation which enables more accurate quantitative glucose measurements continuously. An in vivo time trace of implanted venous sensors demonstrated glucose values that correlated well with the discrete measurements of blood samples on a benchtop point-of-care sensor-based instrument. The raw measured currents from the implanted glucose sensors over 7 h time periods were converted to glucose concentration through use of both a one-point in vivo calibration and a calibration curve obtained in vitro within a bovine serum solution. Control sensors, assembled without NO release functionality, exhibit distinctive surface clotting over the 7 h in vivo implantation period.     

A Telemetry-Instrumentation System for Long-Term Implantable Glucose and Oxygen Sensors

Abstract

This paper describes the development of a compact, low power, implantable system for in vivo monitoring of oxygen and glucose concentrations. The telemetry-instrumentation system consists of two amperometric sensors: one oxygen and one glucose biosensor and two potentiostats for biasing the sensors, an instrumentation amplifier to subtract and amplify sensor output signals, and a signal transmitter subunit to convert and transmit glucose dependent signal from the sensors to a remote data acquisition system. The system produces a unipolar glucose dependent voltage in the range of 1 to 3.6 V which is converted to a frequency and then transmitted using a frequency-modulated (FM) oscillator. Initial tests were performed on an open model electronic circuit using resistors to simulate sensor outputs in the 10 to 1000 nA range. Further in vitro evaluation of the system was conducted with a compact printed circuit board embedded in silicone elastomer, entirely submerged in buffer solution using actual sensors. The test results indicated satisfactory operation of the system in simulated implantation conditions for seven days. Response curve of transmitted signal vs glucose concentration was obtained. The results of the in vitro evaluation of the telemetry system permits its subcutaneous implantation in an animal model.     

Novel CeO2–CuO-decorated enzymatic lactate biosensors operating in low oxygen environments

The detection of the lactate level in blood plays a key role in diagnosis of some pathological conditions including cardiogenic or endotoxic shocks, respiratory failure, liver disease, systemic disorders, renal failure, and tissue hypoxia. Here, we described for the first time the use of a novel mixed metal oxide solution system to address the oxygen dependence challenge of first generation amperometric lactate biosensors. The biosensors were constructed using ceria-copper oxide (CeO₂–CuO) mixed metal oxide nanoparticles for lactate oxidase immobilization and as electrode material. The oxygen storage capacity (OSC, 492 μmol-O₂/g) of these metal oxides has the potential to reduce the oxygen dependency, and thus eliminate false results originated from the fluctuations in the oxygen concentration. In an effort to compare the performance of our novel sensor design, ceria nanoparticle decorated lactate sensors were also constructed. The enzymatic activity of the sensors were tested in oxygen-rich and oxygen-lean solutions. Our results showed that the OSC of the electrode material has a big influence on the activity of the biosensors in oxygen-lean environments. While the CeO₂ containing biosensor showed an almost 21% decrease in the sensitivity in a O₂-depleted solution, the CeO₂–CuO containing electrode, with a higher OSC value, experienced no drop in sensitivity when moving from oxygen-rich to oxygen-lean conditions. The CeO₂–CuO decorated sensor showed a high sensitivity (89.3 ± 4 μA mM⁻¹ cm⁻²), a wide linear range up to 0.6 mM, and a low limit of detection of 3.3 μM. The analytical response of the CeO₂–CuO decorated sensors was studied by detecting lactate in human serum with good selectivity and reliability. The results revealed that CeO₂–CuO containing sensors are promising candidates for continuous lactate detection.     

Amperometric biosensors employing an insoluble oxidant as an interference-removing agent

A strong oxidant membrane is introduced to amperometric biosensors in order to solve the problem associated with interference from readily oxidizable species. The proposed biosensors are in planar format, and are composed of four components, i.e. a base amperometric transducer, an enzyme layer, a protecting membrane, and an oxidant membrane. In this sensor format, interfering species are removed by an oxidation reaction during their diffusion through the oxidant membrane. The oxidant membrane is introduced by dispensing a mixture of an oxidant and a polymer matrix as dissolved in an organic solvent, and thus, could be easily adapted to mass fabrication of miniature biosensors. In this work, several different reagents are examined as oxidants: BaO₂, CeO₂, MnO₂ and PbO₂. Of these, PbO₂ is shown to yield biosensors with the best performance, in terms of reducing interfering signals. Two different matrix systems are devised for use in formulating oxidant membranes: hydrophilic polyurethane (HPU) and cellulose acetate incorporating poly(ethylene glycol) (CA/PEG). While the CA/PEG-type sensor displays better sensitivity and faster response behavior, the HPU-type is shown to exhibit more pronounced interference-removing ability. The analytical utility of the proposed oxidant membrane is demonstrated by fabricating amperometric glucose and creatinine sensors as the model biosensor systems, and by investigating their response characteristics.     

Selectivity coefficients for amperometric sensors

Selectivity coefficients (k(amp)(ij)) are introduced as well-defined measures of the selectivity of amperometric sensors. For any amperometric device the total current response in the presence of interfering species (j) can be described by the general equation; i(t) = K(C(i) + Sigmak(amp)(ij)C(j)), where i is the target analyte. Equations are derived for k(amp)(ij) under different conditions common in amperometric sensing. Factors influencing the selectivity coefficients of amperometric probes, including the recognition and transduction mechanisms, the controlled-potential operating technique, and the transducer geometry are discussed. The selectivity coefficient strategy is illustrated experimentally with two widely used applications of amperometric devices, the anodic monitoring of dopamine at Nafion-coated electrodes and the biosensing of glucose at glucose-oxidase enzyme probes. It is anticipated that the selectivity coefficient strategy will become widespread in amperometric devices, and for chemical sensors in general, and will not be limited only to potentiometric probes.     

A Highly Sensitive Nonenzymatic Glucose Sensor Based on Carbon Electrode Amplified with PdxCuy Catalyst

Approximately global Pd and Pd₉₄Cu₆ alloy nano catalysts of average diameter 10.5 and 5.9 nm respectively, have been synthesized hydrothermally by wet chemical reduction and co-reduction methods without addition of any capping agent. X-ray diffraction and various microscopic studies are used to characterize the crystal phase and the morphology of the catalysts. Non-enzymatic amperometric glucose sensors based on these synthesized catalyst materials are tested and compared in alkali at different potentials by cyclic voltammetry and chronoamperometry. The sensors characterized by fixed potential chronoamperometry are found to be sufficiently sensitive to glucose at different negative potentials like −0.65 V, −0.40 V, −0.10 V with respect to Hg/HgO electrode (E⁰≈0.1 V), where the reactions of glucose oxidation are different. The sensor constructed with Pd₉₄Cu₆ nanocatalyst shows an outstanding sensitivity of 10.1 mA cm⁻² mM⁻¹ which is considerably higher than that constructed with similarly synthesized Pd nanoparticles at any potential and that found in the literature of Pd based glucose sensors. The lower detection limit and response time obtained with Pd₉₄Cu₆ nanoparticles are 10 μM and 3 s respectively. These sensors also exhibit high specificity to glucose and significant anti-interference property against some common species like ascorbic acid (AA), uric acid (UA) and some monosaccharides whose interfering effects are found to decrease with decrease of potential of glucose oxidation. The electrocatalytic ability of the synthesized Pd and Pd₉₄Cu₆ nanoparticles toward glucose oxidation has also found promising in blood sample at different potentials.     

Glucose biosensor based on graphite electrodes modified with glucose oxidase and colloidal gold nanoparticles

The electrochemistry of glucose oxidase (GOx) immobilized on a graphite rod electrode modified by gold nanoparticles (Au-NPs) was studied. Two types of amperometric glucose sensors based on GOx immobilized and Au-NPs modified working electrode (Au-NPs/GOx/graphite and GOx/Au-NPs/graphite) were designed and tested in the presence and the absence of N-methylphenazonium methyl sulphate in different buffers. Results were compared to those obtained with similar electrodes not containing Au-NPs (GOx/graphite). This study shows that the application of Au-NPs increases the rate of mediated electron transfer. Major analytical characteristics of the amperometric biosensor based on GOx and 13 nm diameter Au-NPs were determined. The analytical signal was linearly related to glucose concentration in the range from 0.1 to 10 mmol L^?1. The detection limit for glucose was found within 0.1 mmol L^?1 and 0.08 mmol L^?1 and the relative standard deviation in the range of 0.1–100 mol L^?1 was 0.04–0.39%. The τ_1/2 of V _max characterizes the storage stability of sensors: this parameter for the developed GOx/graphite electrode was 49.3 days and for GOx/Au-NPs/graphite electrode was 19.5 days. The sensor might be suitable for determination of glucose in beverages and/or in food.     

Studies on optimization of a platinum catalyst and porphine modified, pyrolytic graphite, amperometric, glucose sensor by sequential level elimination experimental design

A new amperometric glucose sensor based on the glucose oxidase immobilized on pyrolytic graphite (PG) modified with tetraammineplatinum(II) chloride (TAPtCl) and 5,10,15,20-tetrakis (4-methoxy-phenyl)-21H,23H-porphine cobalt(II) (TMPPCo) as well as Nafion was studied. The performances amongst the glucose sensors with or without TAPtCl or/and TMPPCo measured with oxygen present in the solution were compared. The compositions of the membranes of the glucose sensors were optimized by a new orthogonal experimental design technique–sequential level elimination method according to chemometric approaches. Our studies show that the prepared sensor with optimal membrane composition in this study gives satisfactory performance in terms of long-term stability, fast amperometric response, good detection limits and satisfactory recovery. The study provides a useful basis for developing other sensors with corresponding optimal membranes.     

An implantable biochip to influence patient outcomes following trauma-induced hemorrhage

Following hemorrhage-causing injury, lactate levels rise and correlate with the severity of injury and are a surrogate of oxygen debt. Posttraumatic injury also includes hyperglycemia, with continuously elevated glucose levels leading to extensive tissue damage, septicemia, and multiple organ dysfunction syndrome. A temporary, implantable, integrated glucose and lactate biosensor and communications biochip for physiological status monitoring during hemorrhage and for intensive care unit stays has been developed. The dual responsive, amperometric biotransducer uses the microdisc electrode array format upon which were separately immobilized glucose oxidase and lactate oxidase within biorecognition layers, 1.0–5.0 μm thick, of 3 mol% tetraethyleneglycol diacrylate cross-linked p(HEMA-co-PEGMA-co-HMMA-co-SPA)-p(Py-co-PyBA) electroconductive hydrogels. The device was then coated with a bioactive hydrogel layer containing phosphoryl choline and polyethylene glycol pendant moieties [p(HEMA-co-PEGMA-co-HMMA-co-MPC)] for indwelling biocompatibility. In vitro cell proliferation and viability studies confirmed both polymers to be non-cytotoxic; however, PPy-based electroconductive hydrogels showed greater RMS 13 and PC12 proliferation compared to controls. The glucose and lactate biotransducers exhibited linear dynamic ranges of 0.10–13.0 mM glucose and 1.0–7.0 mM and response times (t ₉₅) of 50 and 35–40 s, respectively. Operational stability gave 80% of the initial biosensor response after 5 days of continuous operation at 37 °C. Preliminary in vivo studies in a Sprague–Dawley hemorrhage model showed tissue lactate levels to rise more rapidly than systematic lactate. The potential for an implantable biochip that supports telemetric reporting of intramuscular lactate and glucose levels allows the refinement of resuscitation approaches for civilian and combat trauma victims.     

An amperometric glucose biosensor based on the immobilization of glucose oxidase on the CuGeO3 nanowire modified electrode

A new amperometric biosensor based on adsorption of glucose oxidase (GOx) at the CuGeO₃ nanowires (NW) modified glassy carbon electrode (GCE) is reported in this article. The properties of the biosensor were characterized by Fourier transform infrared spectroscopy (FTIR), electrochemical impedance spectroscopy (EIS) and electrochemical techniques. The GCE modified with the CuGeO₃ NW/GOD showed excellent electrocatalytical response to the oxidation of glucose. Different parameters including GOx concentration, working potential and pH of supporting electrolyte that governed the analytical performance of the biosensor have been studied in detail and optimized. The biosensor was applied to detect glucose with a linear range of 0.5 to 7 mM. The biosensor exhibited excellent reproducibility and stability.     

Mediator-free amperometric determination of glucose based on direct electron transfer between glucose oxidase and an oxidized boron-doped diamond electrode

A mediator-free glucose biosensor, termed a “third-generation biosensor,” was fabricated by immobilizing glucose oxidase (GOD) directly onto an oxidized boron-doped diamond (BDD) electrode. The surface of the oxidized BDD electrode possesses carboxyl groups (as shown by Raman spectra) which covalently cross-link with GOD through glutaraldehyde. Glucose was determined in the absence of a mediator used to transfer electrons between the electrode and enzyme. O₂ has no effect on the electron transfer. The effects of experimental variables (applied potential, pH and cross-link time) were investigated in order to optimize the analytical performance of the amperometric detection method. The resulting biosensor exhibited fast amperometric response (less than 5 s) to glucose. The biosensor provided a linear response to glucose over the range 6.67×10⁻⁵ to 2×10⁻³ mol/L, with a detection limit of 2.31×10⁻⁵ mol/L. The lifetime, reproducibility and measurement repeatability were evaluated and satisfactory results were obtained.     

Glucose Biosensor Using Glucose Oxidase and Electrospun Mn2O3‐Ag Nanofibers

The highly porous Mn₂O₃‐Ag nanofibers were fabricated by a facile two‐step procedure (electrospinning and calcination). The structure and composition of the Mn₂O₃‐Ag nanofibers were characterized by SEM, TEM, XRD, EDX and SAED. The as‐prepared Mn₂O₃‐Ag nanofibers were then employed as the immobilization matrix for glucose oxidase (GOD) to construct an amperometric glucose biosensor. The biosensor shows fast response to glucose, high sensitivity (40.60 µA mM^?1 cm^?2), low detection limit (1.73 µM at S/N=3), low K_m,app value and excellent selectivity. These results indicate that the novel Mn₂O₃‐Ag nanfibers‐GOD composite has great potential application in oxygen‐reduction based glucose biosensing.     

Electrochemical Fabrication of Cobalt Oxides/Nanoporous Gold Composite Electrode and its Nonenzymatic Glucose Sensing Performance

A nonenzymatic glucose sensor was successfully established by electrochemically decorating cobalt oxides (CoO_x) on a nanoporous gold electrode (NPG) using cobalt hexacyanoferrate (CoHCF) as a precursor. It exhibited high sensitivity and long‐term stability as well as satisfactory quantification of glucose concentration in human serum samples. The morphology and surface analysis of the resulting CoO_x/NPG were carefully characterized. Two detection methods, cyclic voltammetry and amperometry, were employed to evaluate the performance of CoO_x/NPG towards glucose sensing in alkaline solution. Using cyclic voltammetry, at ?0.5 V, the glucose partial oxidation peak current is linear to the glucose concentration up to 14 mM with a sensitivity of 283.7 µA mM^?1 cm^?2. A linear amperometric response at 0.55 V was obtained in the glucose concentration range from 2 µM to 2 mM with a sensitivity of 2025 µA mM^?1 cm^?2 and a response time <3 s.     

Ultrasensitive non-enzymatic glucose sensing at near-neutral pH values via anodic stripping voltammetry using a glassy carbon electrode modified with Pt3Pd nanoparticles and reduced graphene oxide

We describe an anodic stripping voltammetric (ASV) method for glucose sensing that widely expands the typical amperometric i-t response of glucose sensors. The electrode is based on a working electrode consisting of a glassy carbon electrode modified with Pt-Pd nanoparticles (NPs; in an atomic ratio of 3:1) on a reduced graphene oxide (rGO) support. The material was prepared via the spontaneous redox reaction between rGO, PdCl₄ ²⁻ and PtCl₄ ²⁻ without any additional reductant or surfactant. Unlike known Pt-based sensors, the use of Pt₃Pd NPs results in an ultrasensitive ASV approach for sensing glucose even at near-neutral pH values. If operated at a working voltage as low as 0.06 V (vs. SCE), the modified electrode can detect glucose in the 2 nM to 300 μM concentration range. The lowest detectable concentration is 2 nM which is much lower than the LODs obtained with other amperometric i-t type sensing approaches, most of which have LODs at a μM level. The sensor is not interfered by the presence of 0.1 M of NaCl.

We describe an anodic stripping voltammetric method for glucose sensing that widely expands the typical amperometric i-t response of glucose sensors (2 nM to 300 μM). The electrode is based on a glassy carbon electrode modified with Pt-Pd nanoparticles on a reduced graphene oxide (rGO) support.

     

Polymer-Complex Mediated Enzyme Electrodes for Amperometric Determination of Glucose

The immobilized enzyme chemically modified solid-state electrodes based on bilayer-film coating for amperometric determination of glucose have been fabricated and their sensor characteristics have been examined. The electrode substrate was coated with two kinds of polymeric films in a bilayer state, that is, System I: first with the cobalt tetrakis(o-aminophenyl)porphyrin polymer (poly-CoTAPP) film and then with an enzyme film consisting of bovine serum albumin and glucose oxidase (GOx), and System II: first with the Ru(NH₃)³⁺ ₆-containing montmorillonite clay film and then with the GOx enzyme film. The glucose concentration could be monitored by measuring the currents corresponding to the O₂ reduction and the H₂O₂ reduction which are electrocatalyzed by the poly-CoTAPP film (System I) and the clay film (System II), respectively. The reproducible relationship between glucose concentration and sensor output was obtained for both systems with a dynamic range of ～ 1-100 mM (for System I as an electrochemical detector for a flow injection analysis) and ～0.4-4 mM (for System II). In addition, the sensors showed long-term stability (more than 1 and 2 months in System I and System II, respectively) and relatively rapid response (response times of System I and System II are ? 5-10 and 40-60 s, respectively).     

A Multistep Enzyme Sensor for Sucrose Based on S-Layer Microparticles As Immobilization Matrix

Abstract

In this paper we report on the construction principle and performance of an amperometric 3-enzyme sensor for sucrose based on crystalline bacterial cell surface layers (S-layers) as immobilization matrix for the biological components.

Isoporous, crystalline surface layers (S-layers) have been identified as outermost cell envelope layer in many bacteria. Since they are composed of identical protein or glycoprotein subunits with functional groups in well defined positions and orientations, they represent ideal matrices for the controlled and reproducible immobilization of functional macromolecules, as required for the development of biosensors. Apart from single enzyme sensors, which were described earlier, a strikingly simple method for the assembly and optimization of multistep systems was developed. For the fabrication of an amperometric sucrose sensor invertase, mutarotase and glucose oxidase were individually immobilized on S-layer fragments isolated from Clostridium thermohydrosulfuricum L111-69 via aspartic acid as spacer molecules. Subsequently, appropriate mixtures of enzyme loaded S-layer fragments were deposited on a microfiltration membrane and finally, the composite multifunctional sensing layer was sputtered with gold in order to establish a good metal contact. Amperometric sucrose measurements based on H₂O₂ oxidation revealed a high signal level (1 μA^?1/cm^2?mmol sucrose), 5 min response time and a linear range up to 30 mM sucrose as the main characteristics of the S-layer sucrose sensor.     

Continuous glucose monitoring in interstitial fluid using glucose oxidase-based sensor compared to established blood glucose measurement in rats

Glucose monitoring is of importance for success of complex therapeutic interventions in diabetic patients. Its impact on treatment and glycemic control is demonstrated in large clinical trials. Up to eight blood glucose measurements per day are recommended. Notwithstanding, a substantial number of diabetic patients cannot or will not monitor their blood glucose appropriately. Considerable progress in control of disturbed metabolism in diabetic patients can be expected by continuous glucose monitoring. The aim of the study was to evaluate the performance of a new amperometric glucose oxidase-based glucose sensor in vitro and in vivo after subcutaneous implantation into rats.For in vitro testing current output of sensors was measured by exposure to increasing and decreasing glucose concentrations up to 472 mg dL⁻¹ over a time period of 7 days. After subcutaneous implantation of sensors into interscapular region of male rats glucose in interstitial fluid was evaluated and compared to glucose in arterial blood up to 7 days. Hyper- and hypoglycaemia were induced by intravenous application of glucose and insulin, respectively. Current of each implanted sensor was converted into glucose concentration using the first blood glucose measurement only.A change of current with glucose of 0.35 nA mg⁻¹ dL⁻¹ indicates high sensitivity of the sensor in vitro. The response time (90% of steady state) was calculated by approximately 60 s. Test strips for blood glucose measurement as reference for sensor readings was found as an appropriate and rapidly available method in rats by comparison with established hexokinase method in an automated lab analyzer with limits of agreement of +32.8 and −25.7 mg dL⁻¹ in Bland-Altman analysis. In normo- and hypoglycaemic range sensor readings in interstitial fluid correlated well with blood glucose measurements whereas hyperglycaemia was not reflected by the sensor completely when blood glucose was changing rapidly.The data given characterize a sensor with high sensitivity, long term stability and short response time. A single calibration of the sensor is required only in measurement periods up to 7 days. The findings demonstrate that the sensor is a highly promising candidate for assessment in humans.