Actas de congresos
Potentiometric Biosensor For L-ascorbic Acid Based On Ascorbate Oxidase Of Natural Source Immobilized On Ethylene-vinylacetate Membrane
Registro en:
Analytica Chimica Acta. , v. 385, n. 1-3, p. 3 - 12, 1999.
32670
10.1016/S0003-2670(98)00327-4
2-s2.0-0032999632
Autor
Fernandes J.C.B.
Kubota L.T.
Neto G.D.O.
Institución
Resumen
The development of a potentiometric biosensor for L-ascorbic acid based on ascorbate oxidase is described. The enzyme was extracted of the epicarp of Cucumis sativus L. fruit and immobilized in a graphite/epoxy electrode by occlusion in a poly(ethylene-co-vinyl acetate) matrix (EVA 40%m/m). The electrode showed a sub-Nerntian response (slope of 50.3±0.6mV), between 8.0x10-6 and 4.5x10-4moll-1 for ascorbate, when 0.1moll-1 KH2PO4 buffer at pH 5.0 and 25.0°C was employed. The biosensor was continuously tested, during 15 days (about 300 analyses), and no decrease in the response was observed, for vitamin C determination in pharmaceuticals samples, without any previous treatment. The biosensor was stable for at least 1 month when stored in a refrigerator. The relative standard deviation was about 4% compared to the spectrophotometric method. The response time was about 5min at room temperature. Copyright (C) 1999 Elsevier Science B.V. 385 1-3 3 12 Voet, D., Voet, J.G., (1990) Biochemistry, pp. 160F. , Wiley 251F, 536F Tolbert, B.M., Ward, J.B., Ascorbic acid, chemistry, metabolism and uses (1982) Advances in Chemistry, 200. , in: P.A. Seib, B.M. Tolbert (Eds.), American Chemical Society, Washington, DC Clemetson, C.A.B., (1989) Vitamin C, , CRC Press, Boca Raton, FL Nakamura, T., Makino, N., Ogura, Y., (1968) J. Biochem., 64, p. 189 Meyer, T.E., Marchesini, A., Cusanovich, M.A., Tollin, G., (1991) Biochemistry, 30, p. 4619 Cole, J.L., Avigliano, L., Morpurgo, L., Solomon, E.I., (1991) J. Am. Chem. Soc., 113, p. 9080 Maritano, S., Kohzuma, T., Suzuki, S., Marchesini, A., (1996) Phytochemistry, 41, p. 349 Esaka, M., Suzuki, K., Kubota, K., (1985) Agric. Biol. Chem., 49, p. 2955 Stevanato, R., Avigliano, L., Finazzi-Agro, A., Rigo, A., (1985) Anal. Biochem., 149, p. 537 Greenway, G.M., Ongomo, P., (1990) Analyst, 115, p. 1297 Matsumoto, K., Yamada, K., Osajima, Y., (1981) Anal. Chem., 53, p. 1974 Bin Saari, N., Fujita, S., Miyazoe, R., Okugawa, M., (1995) J. Food Biochem., 19, p. 321 Głab, S., Koncki, R., Kopczewska, E., Wałcerz, I., Hulanicki, A., (1994) Talanta, 41, p. 1201 Inniello, R.M., Yacynych, A.M., (1983) Anal. Chim. Acta, 146, p. 249 Alexander, P.W., Joseph, J.P., (1981) Anal. Chim. Acta, 131, p. 103 Et-Jammal, A., Bouklouse, A.A., Patriarche, G.J., (1991) Talanta, 38, p. 929 Rover L., Jr., Garcia, C.A.B., De Oliveira Neto, G., Kubota, L.T., (1998) Anal. Chim. Acta, 366, p. 103 Thomas, J.D.R., (1986) Anal. Chim. Acta, 180, p. 289 Fiedler, U., Růžička, J., (1973) Anal. Chim. Acta, 67, p. 179 Li, J., Shen, G., Yu, R., (1995) Analyst, 120, p. 2259 Li, J., Hu, M., Yu, R., (1996) Sensors and Actuators B, 30, p. 65 Janda, P., Weber, J., Dunsch, L., Lever, A.B.P., (1996) Anal. Chem., 68, p. 960 Tse, Y.H., Janda, P., Lever, A.B.P., (1994) Anal. Chem., 66, p. 384 Moser, O., Kanellis, A.K., (1994) J. Exp. Botany, 45, p. 717 (1995) Catalog Fluka Chemika-Biochemika Analytica, , Fluka Chemie AG Industriestrasse 25 CH-9471 Buchs, Switzerland 156 pp Buck, R.P., Lindner, E., (1994) Pure Appl. Chem., 66, p. 2528 Bailey, P.L., (1980) Analysis with Ion-Selective Electrodes, 2nd Ed., , London Carvalho L.B., Jr., Lima, C.J., Medeiros, P.H., (1981) Phytochemistry, 20, p. 2423 Farver, O., Wherland, S., Pecht, I., (1994) J. Biol. Chem., 269, p. 22933 Garret, R.H., Grisham, C.M., (1995) Biochemistry, , Saunders College Publishing, Orlando Suñer, S.P., (1956) Bioquímica, , editorial Paz Montalvo, Madrid Fernandes, J.C.B., De Oliveira Neto, G., Kubota, L.T., (1998) Anal. Chim. Acta, 366, p. 11 MacCarrone, M., D'Andrea, G., Salucci, M.L., Avigliano, L., Finazzi-Agro, A., (1993) Phytochemistry, 32, p. 795 Gerwin, B., Burstein, S.R., Westley, J., (1974) J. Biol. Chem., 249, p. 2005