Journal
ANALYTICA CHIMICA ACTA
Volume 796, Issue -, Pages 68-74Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2013.08.016
Keywords
Droplet; Microfluidic; Electrochemical detection; Concentration gradient; Enzyme inhibition
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Funding
- National Natural Science Foundation of China [21271127, 61171033]
- Nano-Foundation of Science and Techniques Commission of Shanghai Municipality [12nm0504200]
- China Postdoctoral Science Foundation [2012M510828]
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A simple but robust droplet-based microfluidic system was developed for dose-response enzyme inhibition assay by combining concentration gradient generation method with electrochemical detection method. A slotted-vials array and a tapered tip capillary were used for reagents introduction and concentration gradient generation, and a polydimethylsiloxane (PDMS) microfluidic chip integrated with microelectrodes was used for droplet generation and electrochemical detection. Effects of oil flow rate and surfactant on electrochemical sensing were investigated. This system was validated by measuring dose-response curves of three types of acetylcholinesterase (AChE) inhibitors, including carbamate pesticide, organophosphorus pesticide, and therapeutic drugs regulating Alzheimer's disease. Carbaryl, chlorpyrifos, and tacrine were used as model analytes, respectively, and their IC50 (half maximal inhibitory concentration) values were determined. A whole enzyme inhibition assay was completed in 6 min, and the total consumption of reagents was less than 5 mu L. This microfluidic system is applicable to many biochemical reactions, such as drug screening and kinetic studies, as long as one of the reactants or products is electrochemically active. (C) 2013 Elsevier B.V. All rights reserved.
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