Development of an amperometric biosensing method for the determination of L-fucose in pretreated urine

 
This item is provided by the institution :
University of Ioannina
Repository :
Repository of UOI Olympias
see the original item page
in the repository's web site and access all digital files if the item*
share



2004 (EN)
Development of an amperometric biosensing method for the determination of L-fucose in pretreated urine (EN)

Tsiafoulis, C. G. (EN)

Πανεπιστήμιο Ιωαννίνων. Σχολή Θετικών Επιστημών. Τμήμα Χημείας (EL)
Tsiafoulis, C. G. (EN)

The first amperometric biosensing method for the determination of L-fucose is described. L-Fucose is the objective of much current research, as it is considered as a potential marker for various pathologic disorders. Recombinant L-fucose dehydrogenase, having as cofactor beta-nicotinamide adenine dinucleotide phosphate (NAD(+)P), was cross-linked in a water-soluble photosensitive polymer matrix, that is, polyvinyl alcohol (PVA) modified with styrylpyridinium (SbQ), in the presence of BSA and glutaraldehyde. The resulting membrane was sandwiched between two polycarbonate membranes and was mounted in an amperometric cell. The oxidation of the enzymatically produced NADPH was monitored at a platinum anode at +0.25 V versus a silver pseudoreference electrode in the presence of ferricyanide. The system was fully optimized with respect to various analytical parameters. Regarding to the mechanical properties of the membrane and the storage stability of the immobilized enzyme, various parameters were also optimized. Several methods for the pretreatment of urine samples were investigated. Treatment of the samples with PbO2 found to eliminate the interference effect of various electroactive species exist in urine; optimum incubation time was determined since at prolonged incubation times L-fucose is also affected. Calibration curves for the direct and the mediated monitoring of NADPH were liner over the concentration ranges 0.04-1.0 mM (r(2) = 0.9995) and 0.03-3.0 mM (r(2) = 0.9997) fucose, respectively. The detection limits (S/N 3) were 2 and 1.5 muM fucose, respectively. The R.S.D. of the mediated biosensor is better than 1.5% (n = 10, 0.5 mM fucose). The proposed biosensor correlates well with a reference enzymatic method and exhibits very good working and storage stability. (C) 2004 Elsevier B.V. All rights reserved. (EN)

fucose biosensing method (EN)

Πανεπιστήμιο Ιωαννίνων (EL)
University of Ioannina (EN)

Biosens Bioelectron (EN)

2004

<Go to ISI>://000225009000030

Elsevier (EN)



*Institutions are responsible for keeping their URLs functional (digital file, item page in repository site)