A Conductometric Urea Biosensor by Direct Immobilization of Urease on Pt Electrode

Document Type : Research Article

Authors

Laboratory of Microanalysis, Institute of Biochemistry & Biophysics, University of Tehran, P.O. Box 13145-1384, Tehran, I.R. IRAN

Abstract

Urease was immobilized on platinum electrode both by chemical binding and electropolymerization.The conductometric urea biosensor thus prepared showed a detection limit of 4.9×10-5 M and linear dynamic range from 4.9×10-5 to 5.8×10-3 M for urea concentration when the enzyme is covalently immobilized on Pt electrodes. Conductometric transducers respond to the changes in ionic strength thereby leading to uncontrolled inaccuracies. Such interferences were effectively suppressed by here the use of differential sensor pairs. It was shown that measurements in diluted fluids are possible with the use of areference sensor having no immobilized enzyme.Specifically, the sensor constructed by covalent binding provided more reproducible responses than that prepared by electropolymerization. Namely the former preserved 80% of its initial activity over a period of 25 days. In order to provide more reproducibility,a method was developed for regeneration of the sensor surface so that, the renewed sensor would responded to urea almost as new. 

Keywords

Main Subjects


[1] Komaba, Shinichi; Seyama, Michiko; Momma, Toshiyuki, Osaka Tetsuya, Potentiometric biosensor for urea based on electropolymerized electroinactive polypyrrole; Electrochimica   ActaVolume,   42 (3), p. 383 (1997).
[2] Singhal Rahul, Gambhir Anamika, Pandey, M.K., Annapoorni, S., Malhotra, B.D., Immobilization of urease on poly(N-vinyl carbazole)/stearic acid Langmuir–Blodgett films for application to urea biosensor; Biosensors and Bioelectronics, 17 (8), p. 697 (2002).
[3] Magalhães, Júlia M.C.S., Machado, Adélio A.S.C.,Urea potentiometric biosensor based on urease immobilized on chitosan membranes, Talanta 47 (1), p. 183 (1998).
[4] Seki, Atsushi; Ikeda, Syuu-ichi; Kubo, Izumi; Karube, Isao; Biosensors based on light-addressable potentiometric sensors for urea, penicillin and glucose, Analytica Chemica Acta, 373 (1) p. 9 (1998).
[5] Mandolfo, S., Colasanti, G., Santoro, A., Fustolo, G. Arrigo, G., Bucci, R., Spongano, M., Tetta, C., Imbasciati, E., Clinical evaluation of a new biosensor for on-line urea monitoring, Biosensors and Bioelectronics, 11 (5) p. xx (1996).
[6] Walcerz, I., Koncki, R., Leszczynska, E., Glab, S., Enzyme biosensors for urea determination, Biosensors and Bioelectronics, 11 (5) p. viii (1996).
[7] Baohong, Liu, Renqi, Hu, Jiaqi, Deng; Studies on a potentiometric urea biosensor based on an ammonia electrode and urease, immobilized on a γ, Christa; Cammann, Karl; Knoll, Meinhard; Spener, Friedrich; A disposable biosensor for urea determination in blood based on an ammonium-sensitive transducer, Biosensors and Bioelectronics, 14 (1), p. 33 (1999).
[8] Eggenstein, Claudia; Borchardt, Michaek; Diekmann, Christoph; Gründig, Bernd; Dumschat, Christa; Cammann, Karl; Knoll, Meinhard; Spener, Friedrich; A disposable biosensor for urea determination in blood based on an ammonium-sensitive transducer, Biosensors and Bioelectronics, 14 (1) p. 33, January 1 (1999).
[9] Krawczynski Vel Krawczyk, Tadeusz; Moszczynska, Ma&lz.shtsls; gorzata, Trojanowicz, Marek, Inhibitive determination of mercury and other metal ions by  potentiometric  urea biosensor; Biosensors and Bioelectronics,15 (11-12) p. 681 (2000).
[10] Pandey, Prem C., Singh, Govind, Tetraphenylborate doped polyaniline based novel pH sensor and solid-state urea biosensor, Talanta,  55 (4)p. 773 (2001).
[11] Torbicz, W., Pijanowska, D.G., pH-ISFET based urea biosensor, Sensors and Actuators B: Chemical, volume 44 (1-3)p. 370 (1997).
[12] Boubriak, O.A., Soldatkin, A.P., Starodub, N.F., El'skaya, A.K., Sandrovsky, A.K., Determination of urea in blood serum by a urease biosensor based on an ion-sensitive field-effect transistor, Sensors and Actuators B: Chemical Volume: 27 (1-3) p. 429, (1995).
[13] Zamponi, S., Cicero, B. Lo, Mascini, M., Della Ciana, L., Sacco, S., Urea solid-state biosensor suitable for continuous dialysis control, Talanta, 43 (8) p. 1373 (1996).
[14] Tinkilic,  N.,  Cubuk,  O., Isildak, I., Glucose and urea biosensors based on all solid-state PVC–NH2 membrane electrodes, Analytica Chimica Acta, 452 (1) p. 29 (2000).
[15] de Gracia, J., Poch, M., Martorell, D., Alegret, S., Use of mathematical models to describe dynamic behavior of potentiometric biosensors: comparison of deterministic and empirical approaches to a urea flow-through biosensor, Biosensors and Bioelectronics, 11 (1-2) p. 53 (1996).
[16] Adeloju, S.B., Shaw, S.J., Wallace, G.G., Flow injection biosensor for urea, Biosensors and Bioelectronics, 11 (10) p. viii (1996).
[17] Pizzariello,  Andrea,  Stredanský,  Miroslav, Stredanská, Silvia, Miertuš, Stainslav, Urea biosensor based on amperometric pH-sensing with hematein as a pH-sensitive redox mediator, Talanta 54 (4) p. 763 (2001).
[18] Stred’anský,  Miroslav;  Pizzariello,  Andrea; Stred’anská, Silvia; Miertuš, Stanislav; Ampero-metric pH-sensing biosensors for urea, penicillin, and oxalacetate, Analytica Chimica Acta, 415 (1-2),  p. 151 (2000).
[19] Vostiar, Igor, Tkac, Jan; Sturdik, Ernest; Gemeiner, Peter, Amperometric urea biosensor based on urease and electropolymerized toluidine blue dye as a pH-sensitive redox probe, Bioelectrochemistry, 56 (1-2) p. 113 (2002).
[20] Adeloju, S.B., Shaw, S.J., Wallace, G.G., Pulsed-amperometric detection of urea in blood samples on a conducting polypyrrole-urease biosensor, Analytica Chimica Acta 341 (2-3) p. 155 (1997).
[21] Adeloju, Samuel B., Shaw, Shannon J., Wallace, Gordon G., Polypyrrole-based amperometric flow injection  biosensor for urea, Analytica Chimica Acta, 323 (1-3) p. 107 (1996).
[22] Xie, B., Harborn, U., Mecklenburg, M., Danielsson, B., Miniature thermal biosensor determines urea and lactate, Biosensors and Bioelectronics, 10 (5) p. i-ii (1995).
[23] Bjarnason, Bjarni, Johansson, Peter, Johansson, Gillis, A novel thermal biosensor: evaluation for determination of urea in serum; Analytica Chimica Acta, 372 (3) p. 341 (1998).
[24] Qin, Wei, Zhang, Zhujun; Peng, Youyuan, Plant tissue-based chemiluminescence flow biosensor for urea Analytica Chimica Acta, 407 (1-2) p. 81 (2000).
[25] Jenkins,  Daniel  M.,  Delwiche,  Michael  J., Manometric biosensor for on-line measurement of milk urea, Biosensors and Bioelectronics, 17 (6-7) p. 557 (2002).
[26]  Lee, Won-Yong, Kim, Seung-Ryeol, Kim, Tae-Han, Lee, Kang Shin, Shin, Min-Chol, Park, Je-Kyun, Sol–gel-derived thick-film conductometric biosensor for urea determination in serum; Analytica Chimica Acta, 404 (2) pp. 195 (2000).
[27] Castillo-Ortega, M.M., Rodriguez, D.E., Encinas, J.C., Plascencia, M., Méndez-Velarde, F.A., Olayo, R.; Conductometric uric acid and urea biosensor prepared from electroconductive polyaniline–poly(n-butyl methacrylate) composites, Sensors and ActuatorsB:Chemical, volume85 (1-2) p. 19 (2002).
[28] Sheppard Jr, Norman F., Mears, David J., Guiseppi-Elie, Anthony, Model of an immobilized enzyme conductimetric urea biosensor, Biosensors and Bioelectronics, 11 (10) p. 967 (1996).
[29] Jacobs P., Suls, J., Sansen W., Hombrouckx R., A disposable urea sensor for continuous monitoring of hemodialysis efficiency, ASAIO J., Jul-Sep, 39 (3) 353 (1993).
[30] Mikkelsen, R. S., Rechnitz, A. G., Conductometric transducers for enzyme-based biosensors, Anal.Chem., 61, p. 1737 (1989).
[31] Shul`ga, A. A., Soldatkin, A. P., El`skaya, A.V., Dzyadevich S.V., Patskovsky S.V., Strikha V.I., Thin film conductometric biosensors for glucose and urea determination, Biosensors& Bioelectronics9, p. 217 (1994).
[32] Lawrence, A.J., and Moores, G.R., Conductometry in enzyme studies, Eur. J. Biochem., 24, 538 (1972).
[33] Lawrence, A.J., Conductometric enzyme assays, Eur. J. Biochem., 18, 221 (1971).
[34] Chin, W. T., KroontJe, W., Conductivity method for estimation of urease, Analytical Chemistry, 33 (12) p. 1757 (1961).
[35] Walker, John M., “The Protein Protocols Handbook”, Humana Press Inc., (1996).
[36] Wortington Enzyme Manual, Wortington Bio-chemical Corporation, Free Hold, New Jersey, pp. 339–341 (1972).
[37] Bo Mattiasson and Rajni Kaul, Determination of coupling yields and handling of labile proteins in immobilization technology, in: “Protein immo-bilization fundamentals and applications”, Richard F. Taylor (ed), Chapter 5, P. 161, Marcel Dekker Inc. USA (1991).
[38] Guilbault, G.G., Recommendations for publishing manuscripts on ion selective electrodes, “Ion Selective Electrode Review”, 1, p. 139 Pergamon Press Ltd., UK (1978).
[39] Khodakarami, Hassan, Ghourchian, Hedayat O., PVC-based cerium phosphate membrane exhibiting selectivity for molybdate anions, Iran.