Uricase-adsorbed carbon-felt reactor coupled with a peroxidase-modified carbon-felt-based H2O2 detector for highly sensitive amperometric flow determination of uric acid.
ABSTRACT Uricase (urate oxidase, UOx) was adsorbed onto a porous carbon-felt (CF) surface and the resulting UOx-adsorbed CF (UOx-CF) was successfully used as a column-type enzyme reactor coupled with a peroxidase-adsorbed CF-based bioelectrocatalytic H(2)O(2) flow-detector to fabricate a flow-amperometric biosensor for uric acid. In this flow-biosensor system, H(2)O(2) produced in the UOx-CF reactor was cathodically detected by horseradish peroxidase (HRP) and a thionine (Th)-coadsorbed CF (HRP/Th-CF)-based bioelectrocatalytic flow-detector at -0.05V vs. Ag/AgCl. Various adsorption conditions of the UOx (i.e., pH of the adsorption solution, type and concentration of the buffer used as the adsorption solvent, UOx concentration and adsorption time) and the operational conditions of the UOx-CF and HRP/Th-CF-coupled flow-biosensor (i.e., carrier flow rate and carrier pH) were optimized to obtain highly sensitive, selective and stable peak current responses to uric acid. The analytical performance of the UOx-CF and HRP/Th-CF-coupled flow biosensor for uric acid was as follows: sensitivity, 0.25μA/uM; linear range, 0.3-20μM; lower detection limit, 0.18μM; and sample throughput, ca. 30-90 samples/h. The resulting amperometric flow-biosensor for uric acid allowed the determination of uric acid in highly diluted body fluids (human serum and urine), and the analytical results obtained by the present biosensor were in fairly good agreement with those obtained by conventional enzyme-based spectrophotometry.
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ABSTRACT: We report a new and efficient protocol for rapid electrodeposition of a gold-Prussian blue ((Au-PB)REd) nanocomposite with ultrahigh electroactivity on an Au electrode by cyclic voltammetry in 0.1 M aqueous K2SO4 containing 1 mM K3Fe(CN)6, 1 mM HAuCl4, and 0.1 mM Fe2(SO4)3, and an electrochemical quartz crystal microbalance and scanning electron microscopy were utilized to investigate the electrodeposition. The (Au-PB)REd/Au electrode was then cast-coated with a urate oxidase (UOx)-poly(anilineboronic acid) (PABA)-Pt nanoparticle (PtNP) bionanocomposite and chitosan (CS) for high-performance amperometric biosensing of uric acid (UA) in the dual-potential mode. The UOx-PABA-PtNP bionanocomposite was prepared through chemical oxidation of anilineboronic acid (ABA) by sodium chloroplatinate in the presence of UOx. The thus-fabricated CS/UOx-PABA-PtNP/(Au-PB)REd/Au enzyme electrode worked well under optimized conditions through both oxidation and reduction determination of enzyme-generated H2O2, which responded linearly to UA concentrations from 0.3 μM to 0.65 mM with a sensitivity of 223 μA mM(-1) cm(-2) and a limit of detection (LOD) of 0.2 μM (0.7 V vs. SCE), and from 0.2 μM to 0.25 mM with a sensitivity of 247 μA mM(-1) cm(-2) and a LOD of 0.1 μM (-0.05 V vs. SCE), being superior to most analogues hitherto reported for biosensing of UA.The Analyst 04/2014; · 4.23 Impact Factor