Abstract
A L-ascorbic acid biosensor based on ascorbate oxidase has been developed. The enzyme wasextracted from the mesocarp of cucumber (Cucumis sativus) by using 0.05 mol L -1 phosphate buffer, pH
5.8 containing 0.5 mol L -1 NaCl. After the dialysis versus phosphate buffer
0.05 mol L -1 pH 5.8, the enzyme was immobilized onto nylon net through glutaraldehyde covalent
bond. The membrane was coupled to an O 2 electrode and the yielding reaction monitored by oxygen
depletion at -600 mV using flow injection analysis optimized to 0.1 mol L -1 phosphate buffer pH 5.8, as the
carrier solution and flow-rate of 0.5 mL min -1 . The ascorbic acid calibration curve was linear from 1.2x10 -4 to
1.0x10 -3 mol L -1 . The evaluation of biosensor lifetime leads to 500 injections. Commercial pharmaceutical
samples were analyzed with the proposed method and the results were compared with those obtained by
high-performance liquid chromatography (HPLC).
References
B. A. Fox, A. G. Cameron, Food Science, Nutrition and
Healths, 5th Edition, Edward Arnold, London, 1989, p. 261.
M. J. Villanueva, M. D. Tenorio, M. Sagardoy, A.
Redondo, M. D. Saco, Food Chem. 91(2005)609.
A.Hodgkinson, Oxalic acid in biological and medicine. Aca-
demic Press, London, 1977.
E. J. Oliveira, D. G. Watson, J. Chromatogr. A 764(2001)3.
H. J. Iwase, J. Chromatogr. A 881(2000)327.
A. Rizzolo, A. Brambilla, S. Valsecchi, P. Eccher-Zerbini,
Food Chem. 77(2002)257.
M. Rodríguez-Comsaña, M. S. García-Falcón, J. Siman-
Gándara, Food Chem. 79(2002)141.
W. M. Zeng, F. Martinuzzi, A. MacGregor, J. Pharm.
Biomed. Anal. 36(2005)1107.
L. Fernandez, H. Carrero, Electrochim. Acta
(2005)1233.
J. M. Zen, D. M. Tsai, A. S. Kumar, Electroanal.
(2003)1171.
J. Kulys, E. J. D’Costa, Anal. Chim. Acta 243(1991)173.
J. Kulys, A. Drungiliene, Electroanal. 3(1991)209.
M. Mascini, M. Iannello, G. Palleschi, Anal. Chim. Acta
(1983)135.
L. Macholán, B. Chmlíková, Anal. Chim. Acta 185(1986)187.
S. Uchiyama, Y. Umetsu, Anal. Chim. Acta 255(1991)53.
K. Matsumoto, K. Yamada, Y. Osajima, Anal. Chem.
(1981)1974.
E. Akyilmaz, E. Dinçkaya, Talanta 50 (1999) 87.
O. Fatibello-Filho, I.C. Vieira, J. Braz. Chem. Soc.
(4)(2000)412.
A. Manzoli, I. N. Tomita, F. L. Fertonani, G. Oliveira-
Neto, M. Mascini, H. Yamanaka, Anal. Lett. 37(2004)1823.
M. K. Sezgintürk, E. Dinçkaya, Bios. & Bioelect. 19
(2004) 835.
K. Rekha, M. D. Gouda, M. S. Thakur, N. G. Karanth,
Bios. & Bioelect. 15 (2000) 499.
R. F. Itzhaki, D. M. Gill, Anal. Biochem. 9(1964)401.
H. U. Bergmeyer, M. GrabL, H.E. Walter, Biochemical
reagents for general use. In: H. U. Bergmeyer (Ed.), Meth.
Enz. Anal. Weinheim: VCH Publishers, Inc., 1988, v. II, p. 157.
H. U. Bergmeyer (Ed.), Meth. Enz. Anal. Weinheim:
VCH Publishers, Inc., 1988, v. VI, p. 668.
I. A. Nicolson, R. Macrae, Analyst 109(1984)267.
H. M. A. B. Cardelo, M. A. C. Moraes, L. Cardelo, Alim.
Nutr. 5(1993/94)65.
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