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.

journal_name

J Pharm Biomed Anal

authors

Wang Y,Hasebe Y

doi

10.1016/j.jpba.2011.08.021

subject

Has Abstract

pub_date

2012-01-05 00:00:00

pages

125-32

eissn

0731-7085

issn

1873-264X

pii

S0731-7085(11)00477-8

journal_volume

57

pub_type

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