Novel electrochemical assay for H2O2 determination in aqueous solutions:: A non time-critical method for H2O2 trace level detection

被引:11
作者
Gajovic-Eichelmann, N [1 ]
Bier, FF [1 ]
机构
[1] Fraunhofer Inst Biomed Engn, Dept Mol Bioanalyt & Bioelect, D-14558 Bergholz Rehbrucke, Germany
关键词
H2O2; peroxidase; resorufin; N-acetyl-3,7-dihydroxyphenoxazine; exhaled breath condensate;
D O I
10.1002/elan.200403212
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A two-step method for H2O2 trace level determination has been developed, utilizing, for the first time, the fluorogenic peroxidase substrate, Amplex Red (N-acetyl-3,7-dihydroxyphenoxazine), in conjunction with an electroanalytical detection scheme: Upon peroxidase catalyzed turnover of H2O2 with Amplex Red, the fluorescent and electroactive product resorufin (7-hydroxy-3H-phenoxazin-3-one) was detected amperometrically in a thin-layer flow cell with high selectivity. A linear detection range from 100 nmol L-1 through 1000 nmol L-1 was obtained. The performance was validated using exhaled breath condensates (EBC) with and without H2O2 added, as real samples. Due to the improved chemical stability of resorufin as compared to H2O2, processing larger numbers of EBC samples is greatly facilitated with the new assay, since the enzymatic reaction can be performed up to 18 hours before the actual measurement.
引用
收藏
页码:1043 / 1050
页数:8
相关论文
共 23 条
[1]   Increased hydrogen peroxide and thiobarbituric acid-reactive products in expired breath condensate of asthmatic patients [J].
Antczak, A ;
Nowak, D ;
Shariati, B ;
Krol, M ;
Piasecka, G ;
Kurmanowska, Z .
EUROPEAN RESPIRATORY JOURNAL, 1997, 10 (06) :1235-1241
[2]  
Antczak Adam, 1999, Archivum Immunologiae et Therapiae Experimentalis, V47, P119
[3]   Microplate-based fluorometric methods for the enzymatic determination of L-glutamate: application in measuring L-glutamate in food samples [J].
Chapman, J ;
Zhou, MJ .
ANALYTICA CHIMICA ACTA, 1999, 402 (1-2) :47-52
[4]   THE SCOPOLETIN ASSAY FOR HYDROGEN-PEROXIDE - A REVIEW AND A BETTER METHOD [J].
CORBETT, JT .
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 1989, 18 (04) :297-307
[5]   The use of polymers coupled with metallised electrodes to allow H2O2 detection in the presence of electrochemical interferences [J].
Daly, DJ ;
O'Sullivan, CK ;
Guilbault, GG .
TALANTA, 1999, 49 (03) :667-678
[6]   Increased exhalation of hydrogen peroxide in patients with stable and unstable chronic obstructive pulmonary disease. [J].
Dekhuijzen, PNR ;
Aben, KKH ;
Dekker, I ;
Aarts, LPHJ ;
Wielders, PLML ;
vanHerwaarden, CLA ;
Bast, A .
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 1996, 154 (03) :813-816
[7]   Mediatorless biosensor for H2O2 based on recombinant forms of horseradish peroxidase directly adsorbed on polycrystalline gold [J].
Ferapontova, EE ;
Grigorenko, VG ;
Egorov, AM ;
Börchers, T ;
Ruzgas, T ;
Gorton, L .
BIOSENSORS & BIOELECTRONICS, 2001, 16 (03) :147-157
[8]   ADP-regulation of mitochondrial free radical production is different with complex I- or complex II-linked substrates: Implications for the exercise paradox and brain hypermetabolism [J].
Herrero, A ;
Barja, G .
JOURNAL OF BIOENERGETICS AND BIOMEMBRANES, 1997, 29 (03) :241-249
[9]   Amperometric biosensor for glutamate using Prussian Blue-based "artificial peroxldase" as a transducer for hydrogen peroxide [J].
Karyakin, AA ;
Karyakina, EE ;
Gorton, L .
ANALYTICAL CHEMISTRY, 2000, 72 (07) :1720-1723
[10]   Electrochemical glucose and lactate sensors based on ''wired'' thermostable soybean peroxidase operating continuously and stably at 37 degrees C [J].
Kenausis, G ;
Chen, Q ;
Heller, A .
ANALYTICAL CHEMISTRY, 1997, 69 (06) :1054-1060