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Sniffer-1: a biosensor for the early detection of the oil and fuel leakage in industrial plants

Sniffer-1: a biosensor for the early detection of the oil and fuel leakage in industrial plants Bignetti E.* & Baldini G.° (*Dipartimento di fisica, Parma; °CESI, Milano). INTRODUCTION

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Sniffer-1: a biosensor for the early detection of the oil and fuel leakage in industrial plants

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  1. Sniffer-1: a biosensor for the early detection of the oil and fuel leakage in industrial plants • Bignetti E.* & Baldini G.° • (*Dipartimento di fisica, Parma; °CESI, Milano) INTRODUCTION In thermal power plants some anomalous events such as lubricating-oil or fuel leakage might occur; these fluids spilling on high-temperature surfaces (e.g. pipelines) could cause damage or fire; thus, a specific device should be envisaged in order to early detect the vapors in the ambient air, recognize the relevant volatile compounds and eventually warn the remote-control room operator. To this aim, the equipment should be positioned close to, or even moved around the main plant components, such as the turbine and the burners. In the previous experiments, simulations were carried out in order to collect and identify the dangerous pollutants; we faced up to aromatic compounds, volatile but partially soluble in water; even more interesting, we discovered that their solutions absorb light in the 250-280 nm range and do interfere with the Cytochrome P-450 (2B4-isoform)/P450-Reductase activity assay, inhibiting the reaction in a quantitative mode (Conti et al. (2000) Biosensors 2000, San Diego, P-57, 299). The inhibition mechanism is still under study; however, the evidences above collectively encouraged us to develop a biosensor, as expected. Since the two enzymes have been not yet immobilized succesfully and in a durable way, we moved towards a FIA (flow-injection analysis) strategy. THE DEVICE The final prototype, called 'Sniffer-1', is composed by: a) an electronic processor for the overall control; b) an extracting device to collect pollutants from the surrounding air into distilled water; c) a spectro-photometer to monitor the pollutants level; d) an enzyme-based FIA apparatus for the specific recognition of the pollutants in the water-solutions. FLUORIMETRIC P-450 SYSTEM ASSAY The assay was based on the method of: Burke et al. (1985) Biochem. Pharmacol., 34, 3337, with minor modifications. Top) protocol of reagents required. Right) fluorescence time-course of P450-system-catalyzed Resorufin production, in the absence (a) and the presence of oil extracts (b-e). FUNCTIONAL SCHEME OF SNIFFER-1 A) Reference-water (distilled water circuit). B) Test-water circuit; aromatic volatile compounds can be extracted from the ambient air into distilled water by means of an air pump; test-water 280 nm-wavelength light-absorbance is constantly analyzed by an U.V.-monitor (Bio Rad) connected with an electronic controller (scheda elettronica di controllo). C) Reagents for the enzymatic analysis. D) Cytochrome-P450/Cytochrome-P450 Reductase enzymatic mixture. P1) and P2) peristaltic pumps. E) Electrovalves. MG) Magnetic stirrer. Scheda elettronica) Hitachi H8/3644 controller (8 bit CPU, 32k flash memory, 1k RAM, 1 display, 10 bit A/D converter and 1K EEPROM memory. Note that: A, B, C and D are located in a refrigerated container (4°C peltier). MODE OF ACTION When the U.V.-monitor signal overcomes the threshold, the procedure is automatically activated by the electronic board to carry out a couple of enzymatic assays. This controller sends commands to: the electrovalves, the fluorimeter (Bio Rad), the magnetic stirrer and the peristaltic pump (P2); the aim is to: fill the fluorimeter cuvette, follow the reaction assay and void the cuvette at the end (waste). Fixed reagent amounts are mixed within the cuvette at each assay, either the reference-water or the test-water being the carrier. The controller is programmed to wait for a 1,5 min-loading time and then starts picking the fluorimeter data points up every 5secs. Actually, the mean difference between three data points (at 4, 5 and 6 minutes) and a background value is memorized during the reference assay and compared with the test assay; if the mean-value in the test assay is lower than 50% of the reference then the board sends an alarm. Acknowledgements This research was done within the INFM B1 research line of Parma, under contract P1323 between CESI and University of Parma; this work is part of the activity called 'System Research', assigned to CESI by the Italian Ministry of Industry, Commerce and Handicraft. We are greatly indebted to Giulio Galotta and Raffaele Pellicelli for their skilful technical assistance during the realization of the device.

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