S. V. Dzyadevych

Conductometric tyrosinase biosensor for the detection of diuron, atrazine and its main metabolites.

The determination of diuron, atrazine, desisopropylatrazine (DIA) and desethylatrazine (DEA) were investigated using conductometric tyrosinase biosensor. Tyrosinase was immobilised on the biosensor sensitive part by allowing it to mix with bovine serum albumin (BSA) and then cross-linking in saturated glutaraldehyde (GA) vapour for 30min. The determination of pollutants in a solution was performed by comparison of the output signal (i.e percentage of the enzymatic activity) of the biosensor before and after contact with pollutants. The measurement of the enzymatic activity was performed using 4-chlorophenol, phenol and catechol substrates and response times ranging from 1 to 5min were observed. A 4-chlorophenol substrate was used to detect pesticides. A 30min contact time of the biosensor in the pollutant solution was used. Under the experimental conditions employed, detection limits for diuron and atrazine were about 1ppb and dynamic range of 2.3-2330 and 2.15-2150ppb were obtained for diuron and atrazine, respectively. A relative standard deviation (n=3) of the output signal was estimated to be 5% and a slight drift of 1.5muSh(-1) was observed. The 90% of the enzyme activity was still maintained after 23 days of storage in a buffer solution at 4 degrees C.

Conductometric formaldehyde sensitive biosensor with specifically adapted analytical characteristics

A conductometric enzyme biosensor for determination of formaldehyde in aqueous solutions has been developed using interdigitated thin-film planar electrodes and immobilised alcohol oxidase from Hansenula polymorpha . The biosensor steady-state response was reached after about 1 min. Its dynamic range can vary from 0.05 to 500 mM formaldehyde and depends on the time of enzymatic membrane cross-linking by glutaraldehyde and on the buffer concentration used. The biosensor developed was not completely specific and selective. It demonstrated no response to primary alcohols and other substrates alone. Unfortunately, the response of this biosensor in a mixture of formaldehyde and methanol was decreased in comparison to the one observed for pure formaldehyde, even if no response was obtained with the interfering specie alone. The operational stability was not <20 h and the relative standard deviation appeared to be about 3%. Moreover, the storage stability was more than 1 month.

Novel conductometric biosensor based on three-enzyme system for selective determination of heavy metal ions.

A differential pair of planar thin-film interdigitated electrodes, deposited on a ceramic pad, was used as a conductometric transducer. The three-enzyme system (invertase, mutarotase, glucose oxidase), immobilized on the transducer surface, was used as a bioselective element. The ratio between enzymes in the membrane was found experimentally considering the highest biosensor sensitivity to substrate (sucrose) and heavy metal ions. Optimal concentration of sucrose for inhibitory analysis was 1.25 mM and incubation time in the investigated solution amounted to 10-20 min. The developed biosensor demonstrated the best sensitivity toward ions Hg(2+) and Ag(+). A principal possibility of the biosensor reactivation either by EDTA solution after inhibition with silver ions or by cysteine solution after inhibition with mercury ions was shown.

Application of amperometric biosensors for analysis of ethanol, glucose, and lactate in wine.

This article presents the application of amperometric biosensors based on platinum printed electrodes SensLab and immobilized enzymes, alcohol oxidase, glucose oxidase, and lactate oxidase, for wine analysis. Created devices demonstrate linear response to ethanol, glucose, and lactate within the concentration range 0.3-20 mM, 0.04-2.5 mM, and 0.008-1 mM, respectively. No decrease in ethanol and glucose biosensor activity is revealed during 2 months after fabrication, and the operational stability of the lactate biosensor is sufficient only during 4 days. Developed biosensors showed high selectivity to the substrate and are successfully applied to the analysis of such complex mixtures as wine and must. Good correlation of the results of analysis of different wines and must obtained by amperometric biosensors with immobilized oxidases and traditional methods is shown. Created biosensors can be used as a basis of a commercial device for express analysis of ethanol, glucose, and lactate in wine and must during its fermentation. Application of such devices for quality control in foodstuff industry can have great economical effect because determination by biosensors is less expensive, labor-intensive, and lengthy than traditional methods of analysis.

Highly sensitive electrochemical biosensor for bisphenol A detection based on a diazonium-functionalized boron-doped diamond electrode modified with a multi-walled carbon nanotube-tyrosinase hybrid film

A highly sensitive electrochemical biosensor for the detection of Bisphenol A (BPA) in water has been developed by immobilizing tyrosinase onto a diazonium-functionalized boron doped diamond electrode (BDD) modified with multi-walled carbon nanotubes (MWCNTs). The fabricated biosensor exhibits excellent electroactivity towards o-quinone, a product of this enzymatic reaction of BPA oxidation catalyzed by tyrosinase. The developed BPA biosensor displays a large linear range from 0.01 nM to 100 nM, with a detection limit (LOD) of 10 pM. The feasibility of the proposed biosensor has been demonstrated on BPA spiked water river samples. Therefore, it could be a promising and reliable analytical tool for on-site monitoring of BPA in waste water.

A novel urea conductometric biosensor based on zeolite immobilized urease

A new approach was developed for urea determination where a thin film of silicalite and zeolite Beta deposited onto gold electrodes of a conductometric biosensor was used to immobilize the enzyme. Biosensor responses, operational and storage stabilities were compared with results obtained from the standard membrane methods for the same measurements. For this purpose, different surface modification techniques, which are simply named as Zeolite Membrane Transducers (ZMTs) and Zeolite Coated Transducers (ZCTs) were compared with Standard Membrane Transducers (SMTs). Silicalite and zeolite Beta with Si/Al ratios 40, 50 and 60 were used to modify the conductometric electrodes and to study the biosensor responses as a function of changing zeolitic parameters. During the measurements using ZCT electrodes, there was no need for any cross-linker to immobilize urease, which allowed the direct evaluation of the effect of changing Si/Al ratio for the same type of zeolite on the biosensor responses for the first time. It was seen that silicalite and zeolite Beta added electrodes in all cases lead to increased responses with respect to SMTs. The responses obtained from ZCTs were always higher than ZMTs as well. The responses obtained from zeolite Beta modified ZMTs and ZCTs increased as a function of increasing Si/Al ratio, which might be due to the increased hydrophobicity and/or the acid strength of the medium.

Development of conductometric biosensor array for simultaneous determination of maltose, lactose, sucrose and glucose.

The aim of this work was to develop an array of biosensors for simultaneous determination of four carbohydrates in solution. Several enzyme systems selective to lactose, maltose, sucrose and glucose were immobilised on the surface of four conductometric transducers and served as bio-recognition elements of the biosensor array. Direct enzyme analysis carried out by the developed biosensors was highly sensitive to the corresponding substrates. The analysis lasted 2 min. The dynamic range of substrate determination extended from 0.001 mM to 1.0-3.0mM, and strongly depended on the enzyme system used. An effect of the solution pH, ionic strength and buffer capacity on the biosensors responses was investigated; the conditions of simultaneous operation of all biosensors were optimised. The data on cross-impact of the substrates of all biosensors were obtained; the biosensor selectivity towards possible interfering carbohydrates was tested. The developed biosensor array showed good signal reproducibility and storage stability. The biosensor array is suited for simultaneous, quick, simple, and selective determination of maltose, lactose, sucrose and glucose.

Impedimetric immunosensor based on SWCNT-COOH modified gold microelectrodes for label-free detection of deep venous thrombosis biomarker

Measurement of D-dimer has subsequently become an essential element in the diagnostics of deep vein thrombosis and pulmonary embolism; in this context microelectrodes with an area of 9×10(-4) cm(2) were used to develop impedimetric immunosensor for detecting deep venous thrombosis biomarker (D-dimer). The biosensor is based on functionalized carbon nanotubes (SWCNT-COOH) where the antibody (anti-D-dimer) was immobilized by covalent binding. The electrical properties and the morphology of the biolayer were characterized by electrochemical impedance spectroscopy (EIS), cyclic voltammetry and atomic force spectroscopy (AFM). Impedimetric microimmunosensor allows to obtain sensitivity of 40.1 kΩ μM(-1) and detection limit of 0.1 pg/mL (0.53 fM) with linear range from 0.1 pg/mL to 2 μg/mL (0.53 fM to 0.01 μM). We demonstrate that using carbon nanotubes and microelectrodes, high sensitivity and dynamic range were obtained. The biosensor exhibited a short response time of 10 min. Moreover, the studied immunosensor exhibits good reproducibility (R.S.D. 8.2%, n=4).

Optical fibre biosensors using enzymatic transducers to monitor glucose

The construction and performance of a novel enzyme based optical sensor for in situ continuous monitoring of glucose in biotechnological production processes is presented. Sensitive optical coatings are formed from inorganic?organic hybrid polymers (ORMOCER?s) combined with a flurophore (ruthenium complex) and an enzyme, and applied to lenses, declad polymer optical fibre (POF) and polymer clad silica fibre (PCS). The enzyme, glucose oxidase, catalyzes oxidization of glucose to gluconic acid by depleting oxygen. Oxygen consumption is determined by measuring the fluorescence lifetime of metal organic ruthenium complexes which are quenched by oxygen. The coatings developed were designed to adhere to glass and polymer surfaces, to be compatible with enzymes and ruthenium complexes, and were demonstrated both as double- and single-layer structures. The sensor response to gaseous oxygen, dissolved oxygen and dissolved glucose was measured via fluorescence lifetime changes. A best detection limit of 0.5% (vol) has been determined for gaseous O2 with selected ORMOCER? sensing layers. Glucose concentrations were measured to a detection limit of 0.1 mmol L?1 over a range up to 30 mmol L?1. The sensor was usable for 30 days in a bioreactor. The opto-electronic instrumentation and performance in laboratory bioreactors and in an industrial reactor are evaluated.

Feasibility of application of conductometric biosensor based on acetylcholinesterase for the inhibitory analysis of toxic compounds of different nature

This study was aimed at the development of a conductometric biosensor based on acetylcholinesterase considering the feasibility of its application for the inhibitory analysis of various toxicants. In this paper, the optimum conditions for enzyme immobilization on the transducer surface are selected as well as the optimum concentration of substrate for inhibitory analysis. Sensitivity of the developed biosensor to different classes of toxic compounds (organophosphorus pesticides, heavy metal ions, surfactants, aflatoxin, glycoalkaloids) was tested. It is shown that the developed biosensor can be successfully used for the analysis of pesticides and mycotoxins, as well as for determination of total toxicity of the samples. A new method of biosensor analysis of toxic substances of different classes in complex multicomponent aqueous samples is proposed.