Multienzyme electrochemical array sensor for determination of phenols and pesticides.

The screen-printed four-electrode system was used as the amperometric transducer for determination of phenols and pesticides using immobilised tyrosinase, peroxidase, acetylcholinesterase and butyrylcholinesterase. Acetylthiocholine chloride was chosen as substrate for cholinesterases to measure inhibition by pesticides, hydrogen peroxide served as co-substrate for peroxidase to measure phenols. The compatibility of hydrolases and oxidoreductases working in the same array was studied. The detection of p-cresol, catechol and phenol as well as of pesticides including carbaryl, heptenophos and fenitrothion was carried out in flow-through and steady state arrangements. In addition, the effects of heavy metals (Cu(2+), Cd(2+), Fe(3+)), fluoride (NaF), benzene and dimethylsulphoxide on cholinesterase activities were evaluated. It was demonstrated that electrodes modified with hydrolases and oxidoreductases can function in the same array. The achieved R.S.D. values obtained for the flow system were below 4% for the same sensor and less than 10% within a group of five sensors. For the steady state system, R.S.D.s were approximately twice higher. One assay was completed in less than 6min. The limit of detection for catechol using tyrosinase was equal to 0.35 and 1.7muM in the flow and steady state systems, respectively. On the contrary, lower limits of detection for pesticides were achieved in the steady state system-carbaryl 26nM, heptenophos 14nM and fenitrothion 0.58muM.

Gas chromatography-mass spectrometry with headspace for the analysis of volatile organic compounds in waste water.

Headspace analysis combined with high-resolution gas chromatography and detection by mass spectrometry was evaluated for the analysis of 53 volatile organic compounds (VOCs) in river waters, waste waters and treated water samples down to 0.1 microgl(-1) concentration levels. The conditions optimised included sample thermostatting time and temperature, autosampler parameters and the nature of salt, added to the sample. The pollutions origin and their seasonal rippling have been done. It was shown that the content of VOCs in river water mainly correlates to the content of these compounds in waste waters, which shows the anthropogenic character of the pollutions.

Screen-printed multienzyme arrays for use in amperometric batch and flow systems.

Screen-printing technology for electrode fabrication enables construction of amperometric devices suitable for combination of several enzyme electrodes. To develop a biosensor array for characterisation of wastewaters, tyrosinase and horseradish peroxidase (HRP) or cholinesterase-modified electrodes were combined on the same array. The behaviour of the tyrosinase-modified electrode in the presence of hydrogen peroxide (required co-substrate for the HRP-modified electrode) and acetylthiocholine chloride (required co-substrate for cholinesterase) was studied. Performance of bi-enzyme biosensor arrays in the batch mode and in the flow-injection system are discussed.

Amperometric sensors based on tyrosinase-modified screen-printed arrays.

This paper describes the design, development and characteristics of a tyrosinase (polyphenol oxidase) modified amperometric screen-printed biosensor array, with the enzyme cross-linked in a redox-hydrogel namely the PVI(13)-dmeOs polymer. Two types of Au-screen-printed four-channel electrode arrays, differing in design and insulating layer, were compared and investigated. Au-, graphite-coated-Au- and Carbopack C-coated-Au-surfaces, serving as the basis for tyrosinase immobilisation, were investigated and the performances of the different arrays were evaluated and compared in terms of their electrocatalytic characteristics, as well as operational- and storage stability using catechol as model substrate. It was found that the Carbopack C-coated array was the best choice for tyrosinase immobilisation procedure mainly due to a higher mechanical stability of the deposited enzyme layer, combined with good sensitivity and stability for up to 6 months of use. In the batch mode the biosensors responded linearly to catechol up to 30 muM with limits of detection from 0.14 muM. Parameters from cyclic voltammograms indicated that the reversibility of the direct electrochemical reaction for catechol on the three types of electrode surfaces (no tyrosinase modification) was not the limiting factor for the construction and performance of tyrosinase biosensors.