Daniel Thevenot

Electrochemical biosensors: recommended definitions and classification

Two Divisions of the International Union of Pure and Applied Chemistry (IUPAC), namely Physical Chemistry (Commission 1.7 on Biophysical Chemistry formerly Steering Committee on Biophysical Chemistry) and Analytical Chemistry (Commission V.5 on Electroanalytical Chemistry) have prepared recommendations on the definition, classification and nomenclature related to electrochemical biosensors: these recommendations could, in the future, be extended to other types of biosensors. An electrochemical biosensor is a self-contained integrated device, which is capable of providing specific quantitative or semi-quantitative analytical information using a biological recognition element (biochemical receptor) which is retained in direct spatial contact with an electrochemical transduction element. Because of their ability to be repeatedly calibrated, we recommend that a biosensor should be clearly distinguished from a bioanalytical system, which requires additional processing steps, such as reagent addition. A device that is both disposable after one measurement, i.e. single use, and unable to monitor the analyte concentration continuously or after rapid and reproducible regeneration, should be designated a single use biosensor. Biosensors may be classified according to the biological specificity-conferring mechanism or, alternatively, to the mode of physico-chemical signal transduction. The biological recognition element may be based on a chemical reaction catalysed by, or on an equilibrium reaction with macromolecules that have been isolated, engineered or present in their original biological environment. In the latter cases. equilibrium is generally reached and there is no further, if any, net consumption of analyte(s) by the immobilized biocomplexing agent incorporated into the sensor. Biosensors may be further classified according to the analytes or reactions that they monitor: direct monitoring of analyte concentration or of reactions producing or consuming such analytes; alternatively, an indirect monitoring of inhibitor or activator of the biological recognition element (biochemical receptor) may be achieved. A rapid proliferation of biosensors and their diversity has led to a lack of rigour in defining their performance criteria. Although each biosensor can only truly be evaluated for a particular application, it is still useful to examine how standard protocols for performance criteria may be defined in accordance with standard IUPAC protocols or definitions. These criteria are recommended for authors. referees and educators and include calibration characteristics (sensitivity, operational and linear concentration range, detection and quantitative determination limits), selectivity, steady-state and transient response times, sample throughput, reproducibility, stability and lifetime.

Electrochemical Biosensors: Recommended Definitions and Classification

Two Divisions of the International Union of Pure and Applied Chemistry (IUPAC), namely Physical Chemistry (Commission I.7 on Biophysical Chemistry formerly Steering Committee on Biophysical Chemistry) and Analytical Chemistry (Commission V.5 on Electroanalytical Chemistry) have prepared recommendations on the definition, classification and nomenclature related to electrochemical biosensors; these recommendations could, in the future, be extended to other types of biosensors. An electrochemical biosensor is a self-contained integrated device, which is capable of providing specific quantitative or semi-quantitative analytical information using a biological recognition element (biochemical receptor) which is retained in direct spatial contact with an electrochemical transduction element. Because of their ability to be repeatedly calibrated, we recommend that a biosensor should be clearly distinguished from a bioanalytical system, which requires additional processing steps, such as reagent addition. A device which is both disposable after one measurement, i.e., single use, and unable to monitor the analyte concentration continuously or after rapid and reproducible regeneration should be designated a single use biosensor. Biosensors may be classified according to the biological specificity-conferring mechanism or, alternatively, to the mode of physico-chemical signal transduction. The biological recognition element may be based on a chemical reaction catalysed by, or on an equilibrium reaction with macromolecules that have been isolated, engineered or present in their original biological environment. In the latter cases, equilibrium is generally reached and there is no further, if any, net consumption of analyte(s) by the immobilized biocomplexing agent incorporated into the sensor. Biosensors may be further classified according to the analytes or reactions that they monitor: direct monitoring of analyte concentration or of reactions producing or consuming such analytes; alternatively, an indirect monitoring of inhibitor or activator of the biological recognition element (biochemical receptor) may be achieved. A rapid proliferation of biosensors and their diversity has led to a lack of rigour in defining their performance criteria. Although each biosensor can only truly be evaluated for a particular application, it is still useful to examine how standard protocols for performance criteria may be defined in accordance with standard IUPAC protocols or definitions. These criteria are recommended for authors, referees and educators and include calibration characteristics (sensitivity, operational and linear concentration range, detection and quantitative determination limits), selectivity, steady-state and transient response times, sample throughput, reproducibility, stability and lifetime.

Study and development of multilayer needle-type enzyme-based glucose microsensors

Glucose oxidase (GOD) was covalently coupled to a cellulose acetate (CA) layer, using bovine serum albumin (BSA) and parabenzoquinone (PBQ) linkages. Prior to GOD coupling this CA layer was deposited on the platinum tip of a needle-type sensor and covered with an outer layer of polyurethane (PU). Such microsensors were found to be active, their GOD load reaching 1.6 to 3.0 micrograms mm-2 and their glucose response reaching 1 to 3 microA M-1 mm-2, even when the upper limit of their linear range reached 10-30 mM. Due to the multilayer structure and composition of these microsensors, small anions such as ascorbate were partially discriminated from neutral molecules such as hydrogen peroxide. When implanted subcutaneously in anaesthetized rats, sensor responses correlated correctly with blood glucose concentration but presented sensitivity coefficients significantly different to those determined in vitro: a 2 point calibration procedure was found necessary for in vivo experiments.

BATCH METAL REMOVAL BY PEAT : KINETICS AND THERMODYNAMICS

Peat moss, a natural inexpensive material, is able to play an important rrle in treatment processes of metal-bearing industrial effluents since it adsorbs, complexes or exchanges various metal cations. This paper presents kinetics and thermodynamics of batch metal removal reactions by 50 g/l (dry wt) eutrophic or oligotrophic peat particles using Cu 2+, Cd 2÷, Zn 2÷ and Ni 2÷ concentrations ranging from 0.01 to 100 mM. Metal cation removal reactions are moderately rapid in l0 mM metal unbuffered solutions: the forward kinetic constant ranges between 0.005 and 0.17 M-1 s-1, and equilibrium is reached within about 1 h. Under these conditions of pH (2.2-4.2) and concentrations, apparent binding equilibrium constants were found to range between 2 and 3150 M-1 depending upon the peat origin and the metal cation. In 0-6.5 pH-buffered metal cation solutions, the four cations binding reactions behaved differently demonstrating that metal binding equilibrium constant decrease in the order Ni 2+ > Cu 2+ > Cd 2+ = Zn 2÷. When pH is higher than 6.7, more than 90% of a 10 mM metal cation solution is removed by 50 g/l peat particles and metal binding capacities equal 200 mmol kg-1 dry wt, whatever the metal nature and the peat origin. Except for nickel cation which is very strongly bound to peat, all metal cations are completely released when pH is fixed below 1.5.

Towards continuous glucose monitoring: in vivo evaluation of a miniaturized glucose sensor implanted for several days in rat subcutaneous tissue

SummaryA miniaturized amperometric, enzymatic, glucose sensor (outer diameter 0.45 mm) was evaluated after implantation in the subcutaneous tissue of normal rats. A simple experimental procedure was designed for the long-term assessment of the sensors function which was performed by recording the current during an intraperitoneal glucose load. The sensor was calibrated by accounting for the increase in the current during the concomitant increase in plasma glucose concentration, determined in blood sampled at the tail vein. This made it possible to estimate the glucose concentration in subcutaneous tissue. During the glucose load, the change in subcutaneous glucose concentration followed that in blood with a lag time consistently shorter than 5 min. The estimations of subcutaneous glucose concentration during these tests were compared to the concomitant plasma glucose concentrations by using a grid analysis. Three days after implantation (n=6 experiments), 79 estimations were considered accurate, except for five which were in the acceptable zone. Ten days after implantation (n=5 experiments), 101 estimations were accurate, except for one value, which was still acceptable. The sensitivity was around 0.5 nA mmol−1·l−1 on day 3 and day 10. A longitudinal study on seven sensors tested on different days demonstrated a relative stability of the sensors sensitivity. Finally, histological examination of the zone around the implantation site revealed a fibrotic reaction containing neocapillaries, which could explain the fast response of the sensor to glucose observed in vivo, even on day 10. We conclude that this miniaturized glucose sensor, whose size makes it easily implanted, works for at least ten days after implantation into rat subcutaneous tissue.

ELECTROCHEMICAL BIOSENSORS: RECOMMENDED DEFINITIONS AND CLASSIFICATION*

*A special report on the International Union of Pure and Applied Chemistry, Physical Chemistry Division, Commission I.7 (Biophysical Chemistry), Analytical Chemistry Division, Commission V.5 (Electroanalytical Chemistry).

Trace metal determination in total atmospheric deposition in rural and urban areas

The wet, dry and total atmospheric depositions of some metals (Al, Cd, Cr, Cu, Fe, Na, Pb and Zn) were sampled at two sites and atmospheric fallout fluxes were determined for these locations. This work, led by two different research groups, allowed to reach two main goals: to define a simple analytical procedure to secure accurate shipboard sampling and analysis of atmospheric deposition, and to assess anthropogenic impacts of heavy metals to the environment. The first step about the validation step showed that the prevalent deposition type was dry deposition which represents 40, 60 and 80% for Cd, Cu and Pb, respectively. This prevalence of dry deposition in total atmospheric fallout supported the necessity of funnel wall rinsing which contains 30, 50 and 40% of collected Cd, Cu and Pb, respectively. Moreover, the reproducibility of atmospheric deposition collection was determined. The second step was performed by comparing two sampling sites. A rural sampling site, situated in Morvans regional park (250 km south-east of Paris), was chosen for its isolation from any local and regional contamination sources. Fluxes obtained in this area were compared with those obtained at an urban site (Créteil, suburb of Paris) allowing comparison between urban and rural areas and demonstrating the impact of anthropogenic activities on atmospheric deposition of Cr, Cu and Pb.

Batch copper ion binding and exchange properties of peat

Cupric ion fixation by raw peat is likely involved in both cation exchange with H+, Ca2+, Mg2+ and adsorption-complexation, i.e. fixation of the same equivalent of copper ions and anions (NO3−) without any ion release. The importance of both reactions depends largely on initial copper concentration, peat type and pH. Isotherms of copper (initial concentration ranging between 1 and 20 mM) fixation on two types of peat (eutrophic and oligotrophic peat at 30 g d.w./l at pH ranging between 2 and 4) showed that the higher the initial cupric concentration, the more important is this complexation reaction; over this initial cupric concentration range, ion exchange sites were relatively saturated and reached 308 and 101 mmol/kg d.w. for eutrophic and oligotrophic peat whereas no saturation was found for complexation sites, their capacity attaining up to 74 and 119 mmol/kg d.w., respectively. The apparent equilibrium constant for ion exchange with acid-treated peat (initial pH 4.0, 30 g d.w./l) for various metal binding on both peat sites ranged between 1.1 and 10.8 in 15 mM metallic solutions. The apparent affinity in batch conditions for 5 elements may be compared according to the apparent global equilibrium constants, ranging between 1.1 × 10−6 and 20.2 × 10−6: Pb > Cu > Ca > Mg, Zn for eutrophic peat and Pb > Ca > Cu > Mg, Zn for oligotrophic peat.

Fluorescein diacetate hydrolysis as a measure of microbial activity in aquatic systems: Application to activated sludges

Fluorescein diacetate (FDA) hydrolysis has mainly been used, in soil studies, for measurement of microbial activity and/or for enumeration of bacteria. A protocol is proposed to apply the method to sewage treatment plant activated sludge. The results are compared with values of ETS (electron transport system) activity and oxygen consumption. Unlike ETS activity, FDA hydrolysis is not expected to be proportional to O2 consumption. However, its application to aquatic systems is simpler, less expensive and give results with better reproducibility. FDA hydrolysis may thus allow study of biological activity in situations where extensive surveys in time and/or space are needed.

In Vitro and In Vivo Stability of Electrode Potentials in Needle-Type Glucose Sensors: Influence of Needle Material

Enzymatic glucose sensors are based on the amperometric detection of an oxidable species generated during the oxydation of glucose by glucose oxidase. This measurement usually requires a working electrode (anode), an auxiliary electrode (cathode), and a reference electrode, the function ofthe latter being to keep constant the working potential of the anode, which is responsible for current generation. However, in the needle-type glucose sensors proposed so far, the reference electrode is missing, and its function is performed by the auxiliary electrode. We investigated, in vitro and in vivo in rats, the ability of several cathode-needle materials to behave as a reference electrode in two-electrode glucose sensors, i.e., to present a stable auxiliary electrode potential. In vitro, when glucose concentration was raised from 0 to 30 mM, the auxiliary potential of both gold- and silvercoated sensors presented a cathodic drift, whereas that of silver/silver chloride-coated sensors remained stable. In vivo, during insulin-induced hypoglycemia (5.9–2.4 mM), the auxiliary potentials of all sensors remained stable, whereas during glucose infusion (mean blood glucose concentration 11.2 mM), the auxiliary potentials of both gold- and silver-coated sensors presented an anodic drift, whereas those of silver/silver chloride-coated sensors remained stable. We also indirectly quantified the changes in sensor response induced by variations in the working potential in vitro and in vivo, simulating those that might be produced by a drift in the auxiliary potential. Such changes in the working potential could bring about a 30% unspecific variationin sensor response. We conclude that improvements in sensor analytical characteristics should be obtained with silver/silverchloride—coated cathodes.