Ex-vivo determination of blood glucose by microdialysis in combination with infrared attenuated total reflection spectroscopy

Several biosensors have been developed for continuous monitoring of human blood glucose, which is desirable for insulin-dependent diabetic patients. Developments in the field of quantitative assays using infrared attenuated total reflection spectroscopy allow the determination of metabolites at low concentrations. The microdialysis technique can provide a continuous sampling of extracellular body fluids. As only compounds of low molecular weight are passed on, infrared spectrometric quantitation is eased considerably. Samples were obtained by microdialysis of human blood plasma and aqueous glucose solutions. Multivariate calibration by partial least-squares was evaluated for its analytical performance in ex-vivo blood glucose monitoring. Mean squared prediction errors obtained by cross validation were 5.4 mg/dL for dialysate samples from different patients and 1.3 mg/ dL for dialysates from glucose solutions. Further investigations were carried out to achieve miniaturization of the measuring and detection device. Received: 5 December 1996 / Revised: 20 March 1997 / Accepted: 29 March 1997     

Determination of blood glucose parameter from human blood serum by using a quartz crystal microbalance sensor coated with phthalocyanines compounds

Determining the blood glucose level is important for the prevention and treatment of diabetes mellitus. We developed a sensor system using Quartz Crystal Microbalance (QCM) to determine the blood glucose level from human blood serum. This study consists of two experimental stages: artificial glucose/pure water solution tests and human blood serum tests. In the first stage of the study, the QCM sensor with the highest performance was identified using artificial glucose solution concentrations. In the second stage of the study, human blood serum measurements were performed using QCM to determine blood glucose levels. QCM sensors were coated with phthalocyanines (Pcs) by jet spray method. The blood glucose values of 96 volunteers, which ranged from 71 mg/dL to 329 mg/dL, were recorded. As a result of the study, human glucose values were determined with an average error of 3.25%.     

Continuous glucose monitoring by means of mid-infrared transmission laser spectroscopy in vitro

The continuous surveillance of glucose concentration reduces short-term risks and long-term complications for people with diabetes mellitus, a disorder of glucose metabolism. As a first step towards the continuous monitoring of glucose, reagent-free transmission spectroscopy in the mid-infrared region has been carried out in vitro using a quantum cascade laser and an optical silver halide fiber. A 30 μm gap in the fiber allowed for transmission spectroscopy of aqueous glucose solutions at a wavelength of 9.69 μm, which is specific to a molecular vibration of glucose. A noise-equivalent concentration as low as 4 mg/dL was achieved at an average power of 1.8 mW and an integration time of 50 s. This is among the most precise of glucose measurements using mid-infrared spectroscopy. Even with the very low average laser power of 0.07 mW the sensitivity of previous results (using a fiber optical evanescent field analysis) has been improved upon by almost one order of magnitude. Finally, the impact of potentially interfering substances such as other carbohydrates was analyzed.     

The promise of electrochemical impedance spectroscopy as novel technology for the management of patients with diabetes mellitus

Self-monitoring of blood glucose is the standard of care in management of hyperglycemia among patients with diabetes mellitus. To increase the sensitivity and specificity of current devices, a novel method of detecting glucose using electrochemical impedance spectroscopy (EIS) technology is explored. The enzyme glucose oxidase (GOx) was fixed to gold electrodes and a sine wave of sweeping frequencies was induced using a wide range of concentrations of glucose. Each frequency in the impedance sweep was analyzed for the highest response and R-squared value. The frequency with both factors optimized is specific for the glucose-GOx binding interaction and was determined to be 1.17 kHz in purified solutions in both higher and lower ranges of glucose. The correlation between the impedance response and concentration at the low range of detection (0-100 mg dL(-1) of glucose) was determined to be 3.53 ohm/ln (mg dL(-1)) with an R-squared value of 0.90 with a 39 mg dL(-1) lower limit of detection. The same frequency of 1.17 kHz was verified in whole blood under the same glucose range. The above data confirm that EIS offers a new method of glucose detection as an alternative to current technology in use by patients. Additionally, the unique frequency response of individual markers allows for modulation of signals so that several other markers important in the management of diabetes could be measured with a single sensor.     

A Novel Chronoimpedimetric Glucose Sensor in Real Blood Samples Modified by Glucose‐imprinted Pyrrole‐Aminophenylboronic Acid Modified Screen Printed Electrode

In this study, a molecularly imprinted sensor technology is engineered to detect glucose in real blood samples by chronoimpedimetrically. The imprinting process of glucose (Glc) was carried out by electrochemical polymerization of aminophenylboronic acid (APBA) and pyrrole (Py) by performing cyclic voltammetry (CV). Afterwards, glucose molecule was removed from imprinted surface by 5 % acetic acid to reveal glucose imprinted cavities. Electrochemical Impedance Spectroscopy (EIS) was used to characterize sensor modification steps and glucose removal. Glucose monitoring process was carried out chronoimpedimetrically(CI) for the first time in real blood samples. Calibration curve was prepared between 20–800 mg/dL. The standard deviations of the 18 calibration curves R² were calculated as 0.9866±0.0066 to assess reproducibility. Recovery was calculated by using 105 mg/dL Glc Serum Sample, which was monitored by auto analyzer and into this sample 50 mg/dL Glc added and our sensor response was 147.92±2.43 mg/dL, 98.6±1.62 % (n=5). Non‐imprinted (NIP) sensor gave no signal for the glucose concentration.     

A Microdialysis Probe Coupled with A Miniaturized Thermal Glucose Sensor for In Vivo Monitoring

Abstract

A miniaturized thermal flow injection analysis biosensor has been coupled with a microdialysis probe for continuous subcutaneous glucose monitoring. Thermal biosensors are based on the principle of measuring the heat evolved during enzyme catalysed reactions. The system presented here consists of a miniaturized thermal biosensor with a small column containing coimmibolized glucose oxidase and catalase. The analysis buffer passes through the column at a flow rate of 60μL/min via an 1μL sample loop which is connected to a microdialysis probe.

Invitro results showed constant permeability of the probe and stability of the biosensor response during 24 hours. The response time was 85 sec giving a sample rate of 42 samples/hour.

During a load experiment, the glucose profile in a healthy volunteer was followed both in the subcutaneous tissue and blood using the microdialysis set-up proposed and comparing to blood glucose analyser.     

Fiber-optic flow-through sensor for online monitoring of glucose

A new microdialysis-based glucose-sensing system with an integrated fiber-optic hybrid sensor is presented. Design and dimensions of the cell are adapted for its coupling with commercially available microdialysis techniques, thereby providing a new system for continuous glucose monitoring. The glucose level is detected via oxygen consumption which occurs as a consequence of enzymatic reaction between immobilized glucose oxidase and glucose. The use of gas-permeable Tygon tubing ensures complete and constant air-saturation of the measuring fluid in the cell. Nevertheless, a reference oxygen optode is used to detect and to compensate response changes caused by events like bacterial growth, temperature fluctuations, or failure of the peristaltic pump. In contrast to widely used electrochemical sensors, the response of the microdialysis-based fiber-optic glucose sensor is highly selective, making this sensor approach particularly advantageous for continuous glucose monitoring of patients in intensive care units. The effects of flow rate, pH, temperature, and common interferences on the sensor response are presented and discussed in detail. The sensor is evaluated in vitro using a 3-day continuous test in glucose-spiked plasma. The ability to measure glucose in humans is demonstrated by coupling the flow-through cell and commercially available microdialysis catheter CMA60. A 24-h monitoring test using this setup is successfully applied to a healthy volunteer.     

Infrared spectroscopic analysis of human interstitial fluid in vitro and in vivo using FT-IR spectroscopy and pulsed quantum cascade lasers (QCL): Establishing a new approach to non invasive glucose measurement

Interstitial fluid, i.e. the liquid present in the outermost layer of living cells of the skin between the Stratum corneum and the Stratum spinosum, was analyzed by Fourier transform infrared spectroscopy and by infrared spectroscopy using pulsed quantum cascade infrared lasers with photoacoustic detection. IR spectra of simulated interstitial fluid samples and of real samples from volunteers in the 850-1800cm(-1) range revealed that the major components of interstitial fluid are albumin and glucose within the physiological range, with only traces of sodium lactate if at all. The IR absorbance of glucose in interstitial fluid in vivo was probed in healthy volunteers using a setup with quantum cascade lasers and photoacoustic detection previously described. A variation of blood glucose between approx. 80mg/dl and 250mg/dl in the volunteers was obtained using the standard oral glucose tolerance test (OGT). At two IR wavelengths, 1054cm(-1) and 1084cm(-1), a reasonable correlation between the photoacoustic signal from the skin and the blood glucose value as determined by conventional glucose test sticks using blood from the finger tip was obtained. The infrared photoacoustic glucose signal (PAGS) may serve as the key for a non-invasive glucose measurement, since the glucose content in interstitial fluid closely follows blood glucose in the time course and in the level (a delay of some minutes and a level of approx. 80-90% of the glucose level in blood). Interstitial fluid is present in skin layers at a depth of only 15-50μm and is thus within the reach of mid-IR energy in an absorbance measurement. A non-invasive glucose measurement for diabetes patients based on mid-infrared quantum cascade lasers and photoacoustic detection could replace the conventional measurement using enzymatic test stripes and a drop of blood from the finger tip, thus reducing pain and being a cost-efficient alternative for millions of diabetes patients.     

Non-invasive determination of glucose directly in raw fruits using a continuous flow system based on microdialysis sampling and amperometric detection at an integrated enzymatic biosensor

A non-destructive, rapid and simple to use sensing method for direct determination of glucose in non-processed fruits is described. The strategy involved on-line microdialysis sampling coupled with a continuous flow system with amperometric detection at an enzymatic biosensor. Apart from direct determination of glucose in fruit juices and blended fruits, this work describes for the first time the successful application of an enzymatic biosensor-based electrochemical approach to the non-invasive determination of glucose in raw fruits. The methodology correlates, through previous calibration set-up, the amperometric signal generated from glucose in non-processed fruits with its content in % (w/w). The comparison of the obtained results using the proposed approach in different fruits with those provided by other method involving the same commercial biosensor as amperometric detector in stirred solutions pointed out that there were no significant differences. Moreover, in comparison with other available methodologies, this microdialysis-coupled continuous flow system amperometric biosensor-based procedure features straightforward sample preparation, low cost, reduced assay time (sampling rate of 7 h⁻¹) and ease of automation.     

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.     

Continuous monitoring of unbound flomoxef levels in rat blood using microdialysis and its new pharmacokinetic analysis

Microdialysis sampling method coupled to high-performance liquid chromatography with UV detection was applied to continuous monitoring of in vivo unbound flomoxef concentration in rat blood. By comparison with ultrafiltration method, it was demonstrated that it gave reliable results for the unbound drug monitoring in blood. Furthermore, a new method was presented for the calculation of pharmacokinetic parameters from the data obtained by the microdialysis method.     

Noninvasive determination of blood constituents using an array of modulated laser diodes and a photoacoustic sensor head

Using photoacoustic laser spectroscopy, the noninvasive determination of blood constituents like hemoglobin and glucose is feasible. The aim of our investigations is the development of a sensor which is suitable for continuously noninvasive monitoring of blood glucose concentrations in diabetic patients. For this purpose a photoacoustic sensor head was developed and coupled via an optical fiber bundle to an array of 8 laser diodes emitting at various wavelengths in the near infrared region. Applying a special modulation scheme, the tiny changes of the absorption coefficient of whole blood caused by the variations of blood glucose concentrations could be measured. A resolution of 70 mg/dl was achieved, a value which is already close to the clinical requirements for a continuously working glucose sensor.     

Noninvasive determination of blood constituents using an array of modulated laser diodes and a photoacoustic sensor head

Using photoacoustic laser spectroscopy, the noninvasive determination of blood constituents like hemoglobin and glucose is feasible. The aim of our investigations is the development of a sensor which is suitable for continuously noninvasive monitoring of blood glucose concentrations in diabetic patients. For this purpose a photoacoustic sensor head was developed and coupled via an optical fiber bundle to an array of 8 laser diodes emitting at various wavelengths in the near infrared region. Applying a special modulation scheme, the tiny changes of the absorption coefficient of whole blood caused by the variations of blood glucose concentrations could be measured. A resolution of 70 mg/dl was achieved, a value which is already close to the clinical requirements for a continuously working glucose sensor.     

Variable Selection Towards Classification of Digital Images: Identification of Altered Glucose Levels in Serum

Identification of altered glucose levels in serum is the main indicator for diabetes, where control levels are classed as <100?mg/dL, and altered levels are classified as pre-diabetic (100–125?mg/dL) or diabetic (>125?mg/dL). Herein, we propose a method to identify control, pre-diabetic, or diabetic simulated and real-world samples based on their glucose levels using classification-based variable selection algorithms [successive projections algorithm (SPA) or genetic algorithm (GA)] coupled to linear discriminant analysis (SPA-LDA and GA-LDA) towards analyzing red–green–blue digital images. Images were recorded after glucose enzymatic reaction, whereby 250?μL of reactant content of samples were captured by using a common cell phone camera. Processing was applied to the images at a pixel level, where 72.2% of the pixels were correctly classified as control, 79.2% as pre-diabetic, and 90.9% as diabetic using SPA-LDA algorithm; and 76.8% as control, 81.4% as pre-diabetic, and 91.7% as diabetic using GA-LDA algorithm in the validation set containing nine simulated samples. Eight real-world samples were measured as an external test set, where the accuracy using GA-LDA was found to be 92%, with sensitivities ranging from 70% to 100 and specificities ranging from 90% to 99%. This method shows the potential of variable selection techniques coupled with digital image analysis towards blood glucose monitoring.     

Monitoring of glucose in biological fluids by fourier-transform infrared spectrometry with a cylindrical internal reflectance cell

The feasibility of using internal reflectance infrared spectrometry to monitor changing glucose concentrations in biological fluids is described. A FTIR spectrophotometer and a cylindrical internal reflectance cell are used to obtain spectra of glucose in normal saline solutions, blood serum, and cerebral spinal fluid. The slopes obtained for the standard additions of glucose to each of these solutions compare well and indicate that although the changes in absorbance are small, they are quantifiable. The working range was 0–8 mg ml^?1, which brackets normal blood glucose concentrations.     

Determination of Unbound Ampicillin in Rat Blood by Microdialysis and Liquid Chromatography

ABSTRACT

An in vivo microdialysis sampling method coupled to high-performance liquid chromatography has been applied for continuous monitoring of unbound ampicillin in rat blood. A microdialysis probe was inserted into the jugular vein/right atrium of Sprague-Dawley rats, and doses of 100 and 200 mg/kg ampicillin were then administered via the femoral vein. Dialysates were collected and directly injected into a liquid chromatographic system. Isocratic elution of ampicillin was achieved within 10 min using the liquid chromatographic system. The chromatographic mobile phase consisted of methanol-100 mM monosodium phosphoric acid (25:75, v/v, pH 5.5). The wavelength of the UV detector was set at 230 nm. The calibration curves from 0.25 to 50 μg/ml were linear with correlation coefficients of 0.995. The method provides a simple technique for rapid analysis of unbound ampicillin in rat blood for used in pharmacokinetic study.     

Using blood hemoglobin for blood analysis

This paper demonstrates that the spectrophotometric properties of blood hemoglobin (Hb) can be used for the direct determination of biochemical compounds in blood. Glucose is used as a model, but the methodology can be applied to many other compounds (only a previous enzymatic reaction producing H(2)O(2) is needed). In order to develop the method, a model relating the Hb absorbance variation during the reaction with the glucose concentration has been developed to provide theoretical support for the method and to predict its application to other compounds. In addition, clear blood samples need to be prepared without pre-treatment and lateral reactions of H(2)O(2) with other blood constituents need to be blocked; this has been achieved with 100 : 1 v/v blood dilution in bi-distilled water and azide addition. The linear response range of the method can be fitted between 2 and 540 mg dL(-1) glucose relative to the original blood sample (RSD about 4%, 70 mg dL(-1)). The analyte concentration can be obtained by an absolute calibration method or by the standard addition method; both have been applied for direct glucose determination in several blood samples and good correlations with those obtained by an automatic analyzer have been obtained.     

Trends of flow injection sample pretreatment approaching the new millennium

The important role of flow injection (FI) techniques for the automation, acceleration and miniaturization of solution handling in sample pretreatment as well as some recent trends in the development of the field are discussed, illustrated mainly by recent achievements in the author’s laboratory, including: (a) sample pretreatment for vapor generation and electrothermal AAS based on sequential injection (SI) techniques, with low reagent consumption and enhanced ruggedness; (b) combination of FI and SI sample pretreatment (filtration, dialysis, gas diffusion, column sorption) with capillary electrophoresis (CE) giving enhanced reproducibility and efficiency; (c) application of on-line microdialysis in in vivo monitoring of blood glucose in test animals and (d) application of on-line microdialysis and solvent extraction in continuous monitoring of drug dissolution processes with high resolution of process events. Future perspectives of FI sample pretreatment are discussed, emphasizing the improvement in ruggedness of the equipment and methods, the combination and synchronization of different means for liquid propulsion, and the development of miniaturized systems.     

Fabrication of interdigitated high-performance zinc oxide nanowire modified electrodes for glucose sensing

Diabetes is a metabolic disease with a prolonged elevated level of glucose in the blood leads to long-term complications and increases the chances for cardiovascular diseases. The present study describes the fabrication of a ZnO nanowire (NW)-modified interdigitated electrode (IDE) to monitor the level of blood glucose. A silver IDE was generated by wet etching-assisted conventional lithography, with a gap between adjacent electrodes of 98.80 μm. The ZnO-based thin films and NWs were amended by sol–gel and hydrothermal routes. High-quality crystalline and c-axis orientated ZnO thin films were observed by XRD analyses. The ZnO thin film was annealed for 1, 3 and 5 h, yielding a good-quality crystallite with sizes of 50, 100 and 110 nm, and the band gaps were measured as 3.26, 3.20 and 3.17 eV, respectively. Furthermore, a flower-modeled NW was obtained with the lowest diameter of 21 nm. Our designed ZnO NW-modified IDE was shown to have a detection limit as low as 0.03 mg/dL (correlation coefficient = 0.98952) of glucose with a low response time of 3 s, perform better than commercial glucose meter, suitable to instantly monitor the glucose level of diabetes patients. This study demonstrated the high performance of NW-mediated IDEs for glucose sensing as alternative to current glucose sensors.     

Recent progress in analytical instrumentation for glycemic control in diabetic and critically ill patients

Implementing strict glycemic control can reduce the risk of serious complications in both diabetic and critically ill patients. For this reason, many different analytical, mainly electrochemical and optical sensor approaches for glucose measurements have been developed. Self-monitoring of blood glucose (SMBG) has been recognised as being an indispensable tool for intensive diabetes therapy. Recent progress in analytical instrumentation, allowing submicroliter samples of blood, alternative site testing, reduced test time, autocalibration, and improved precision, is comprehensively described in this review. Continuous blood glucose monitoring techniques and insulin infusion strategies, developmental steps towards the realization of the dream of an artificial pancreas under closed loop control, are presented. Progress in glucose sensing and glycemic control for both patient groups is discussed by assessing recent published literature (up to 2006). The state-of-the-art and trends in analytical techniques (either episodic, intermittent or continuous, minimal-invasive, or noninvasive) detailed in this review will provide researchers, health professionals and the diabetic community with a comprehensive overview of the potential of next-generation instrumentation suited to either short- and long-term implantation or ex vivo measurement in combination with appropriate body interfaces such as microdialysis catheters.