Development and characterization of a microfluidic glucose sensing system based on an enzymatic microreactor and chemiluminescence detection  

作　　者：MOON B.-U. de VRIES M.G. WESTERINK B.H.C. VERPOORTE E. 

机构地区：[1]Biomonitoring and Sensoring, Department of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands [2]Pharmaceutical Analysis, Department of Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands [3]Brains-on-Line, PO Box 4030, 9713AV Groningen, The Netherlands

出　　处：《Science China Chemistry》2012年第4期515-523,共9页中国科学（化学英文版）

摘　　要：Chemiluminescence detection was developed as an alternative to amperometric detection for glucose analysis in a portable, microfluidicsbased continuous glucose monitoring system. Amperometric detection allows easy determination of hydrogen peroxide, a product of the glucose oxidasecatalyzed reaction of glucose with oxygen, by oxidation at a microelectrode. However, （micro）electrodes in direct contact with physiological sample are subject to electrode fouling, which leads to signal drift, decreased reproducibility and shortened detector lifetimes. Moreover, there are a few species present in the body （e.g. ascorbic acid, uric acid） which can undergo oxidation at the same applied potential as hydrogen peroxide. These species can thus inter- fere with the glucose measurement, reducing detection specificity. The rationale for exploring chemiluminescence as opposed to amperometric detection is thus to attempt to improve the lifetime and reproducibility of glucose analysis for monitoring purposes, while reducing interference caused by other chemicals in the body. The study reported here represents a first step in this direction, namely the realization of a microfluidic device with integrated silicon photodiode for chemiluminescence detection of glucose. This microflow device uses a chaotic mixing approach to perform enzymatic conversion of glucose, followed by reaction of the hydrogen peroxide produced with luminol to produce light at 425 nm. The chemil reaction is catalyzed by horseradish peroxidase in the presence of iodophenol. The performance of the fabricated chip was characterized to establish optimal reaction conditions with respect to sample and reagent flow rates, pH, and concentrations. A linear calibra- tion curve was obtained for current response as a function of glucose concentration in the clinically relevant range between 2 and 10 mM, with a sensitivity of 39 pA/mM （R = 0.9963, one device, n = 3） and a limit of detection of 230 ktM （S/N - 3）.Chemiluminescence detection was developed as an alternative to amperometric detection for glucose analysis in a portable, microfluidics-based continuous glucose monitoring system. Amperometric detection allows easy determination of hydrogen peroxide, a product of the glucose oxidase-catalyzed reaction of glucose with oxygen, by oxidation at a microelectrode. However, (micro)electrodes in direct contact with physiological sample are subject to electrode fouling, which leads to signal drift, decreased reproducibility and shortened detector lifetimes. Moreover, there are a few species present in the body (e.g. ascorbic acid, uric acid) which can undergo oxidation at the same applied potential as hydrogen peroxide. These species can thus interfere with the glucose measurement, reducing detection specificity. The rationale for exploring chemiluminescence as opposed to amperometric detection is thus to attempt to improve the lifetime and reproducibility of glucose analysis for monitoring purposes, while reducing interference caused by other chemicals in the body. The study reported here represents a first step in this direction, namely the realization of a microfluidic device with integrated silicon photodiode for chemiluminescence detection of glucose. This microflow device uses a chaotic mixing approach to perform enzymatic conversion of glucose, followed by reaction of the hydrogen peroxide produced with luminol to produce light at 425 nm. The chemiluminescence reaction is catalyzed by horseradish peroxidase in the presence of iodophenol. The performance of the fabricated chip was characterized to establish optimal reaction conditions with respect to sample and reagent flow rates, pH, and concentrations. A linear calibration curve was obtained for current response as a function of glucose concentration in the clinically relevant range between 2 and 10 mM, with a sensitivity of 39 pA/mM (R = 0.9963, one device, n = 3) and a limit of detection of 230 μM (S/N = 3).

关 键 词：microfluidic chip – glucose sensing system – enzymatic microreactor – chemiluminescence – chaotic mixing 

分 类 号：R446.11[医药卫生—诊断学]