Amperometric biosensor based on reductive H2O2 detection using pentacyanoferrate-bound polymer for creatinine determination

Abstract

Pentacyanoferrate-bound poly(1-vinylimidazole) (PVI[Fe(CN)5]) was selected as a mediator for amperometric creatinine determination based on the reductive H2O2 detection. Creatinine amidohydrolase (CNH), creatine amidohydrolase (CRH), sarcosine oxidase (SOD), peroxidase (POD), and PVI[Fe(CN)5] were crosslinked with poly(ethylene glycol) diglycidyl ether (PEGDGE) on a glassy carbon (GC) electrode for a creatinine biosensor fabrication. Reduction current was monitored at -0.1V in the presence of creatinine and O2. It is revealed that PVI[Fe(CN)5] is suitable as a mediator for a bioelectrocatalytic reaction of POD, since PVI[Fe(CN)5] neither reacts with reactants nor works as an electron acceptor of SOD. The amounts of PVI[Fe(CN)5], PEGDGE, and enzymes were optimized toward creatinine detection. Nafion as a protecting film successfully prevented the enzyme layer from interferences. The detection limit and linear range in creatinine determination were 12μM and 12-500μM (R(2)=0.993), respectively, and the sensitivity was 11mAcm(-2)M(-1), which is applicable for urine creatinine tests. The results of the creatinine determination for four urine samples measured with this proposed method were compared with Jaffe method, and a good correlation was obtained between the results.

Direct Electron Transfer-type Bioelectrocatalysis of Peroxidase at Mesoporous Carbon Electrodes and Its Application for Glucose Determination Based on Bienzyme System

“…The proposed bienzyme system can be applied to the detection of several substrates by just replacing GOD with the corresponding oxidase, such as D-amino acid oxidase, 11 sarcosine oxidase, 12 and glutamate oxidase. 14 In addition, the reduction of H2O2 at such high electrode potentials on PODimmobilized mesoporous carbon electrodes can be utilized as bio-cathodes in biofuel cells in place of multi-copper oxidasebased bio-cathodes, since the activity of the latter bio-cathodes are liable to be inhibited by H2O2 and halogen ions.…”

“…Considering the fact that the composition of POD and an H2O2-generated oxidase may affect the biosensing performance of bienzyme biosensors, 11,12 the ratio of POD to GOD was examined. Figure S4 (Supporting Information) shows the effects of the POD/GOD ratio on the relative current response to 1 mM glucose (ratios from 0.05 to 0.30 at a constant GOD concentration of 5 mg mL -1 ) in the mixed enzyme/reagent solutions for immobilization.…”

“…7,8 MET-type POD sensors have been frequently combined with an H2O2-generating oxidase for determination of the first substrate of the oxidase. [9][10][11][12][13][14][15] One of the issues to be considered for such MET-type POD/oxidase bienzyme-electrode reactions may be that the oxidized form of a mediator generated in the POD reaction frequently works as an electron acceptor of the oxidase in the bienzyme system (due to the dehydrogenase activity), 13 which results in negative interferences.…”

Two low-cost digital camera-based platforms for quantitative creatinine analysis in urine

“…Due to its medical relevance the development of a fast and accurate assay for the determination of creatinine in human serum or urine is required. In recent literature several techniques have been reported for creatinine assay such as HPLC [3], potentiometric biosensors [4,5], electro-analytical [2,6] and enzymatic [7] based methods, tandem mass spectroscopy [8e11], microchip capillary electrophoresis [12e14], strip assay [15,16] and Raman scattering spectroscopy [17]. Typical standard methods for creatinine quantification in urine samples are high performance liquid chromatography (HPLC) and capillary electrophoresis.…”

Plataformas de baixo custo à base de papel para testes imunodiagnósticos e enzimáticos

“…As enzimas oxidases estão sendo amplamente utilizadas na detecção de moléculas em testes amperométricos, fluorimétricos e colorimétricos [24]. Colesterol, glicose, creatinina entre outros analitos podem ser detectados utilizando-se oxidases [37][38][39][40] tanto que a peroxidase é uma das enzimas mais utilizadas na clínica médica [41].…”