There has been urgent demand for rapid, sensitive and cost-effective pesticide assay technologies due to the global attention of environmental and food-safety problems. Acetycholinesterase (AChE)-based electrochemical sensors have attracted significant interest toward this goal. In this contribution, we introduced multiwalled carbon nanotubes (MWNTs) into our sensor design, where they played dual enhancement roles; first is that MWNTs loaded on glassy carbon (GC) electrodes significantly increase surface areas, facilitating the electrochemical polymerization of prussian blue (PB), a redox mediator for the electrochemical oxidation of the enzymatic product, thiocholine (TCh). Second, MWNTs enhance the enzymatic activity of AChE, as manifested by the decreased Michaelis-Menten constant (K(m)). As a result of these two important enhancement factors offered by MWNTs, our electrochemical pesticide sensor exhibited rapid response and high sensitivity toward the detection of a series of pesticides. Moreover, we demonstrated that this sensor was stable, reproducible and selective enough for detection in real samples.
Recombinant Drosophila melanogaster acetylcholinesterase (R-DmAChE), multiwalled carbon nanotubes (MWCNTs), and Prussian blue have been combined for development of a three-electrode biosensor with more rapid responses and higher stability than in our previous study. A new disposable screen-printed electrode (SPE) was developed for rapid detection of organophosphate and carbamate pesticides. After optimization, 10 μg MWCNT and 5 μL enzyme immobilization solution consisting of 0.2% glutaraldehyde, 0.1% Nafion®, 0.2% bovine serum albumin, 0.1 g/L MWCNT, and 1.5 mU R-DmAChE were fixed on each of the R-DmAChE/MWCNT SPEs. The LOD of this biosensor was 0.5 μg/L for pesticide standards of dichlorvos (DDV) and carbofuran. The performance of this biosensor was tested for vegetable and water samples at various spiked levels, and good stability and sensitivity were found. The obtained recoveries were from 82.6 to 110.5% for DDV at levels of 0.5–5 μg/L and 73.4 to 118.4% for carbofuran at 1–10 μg/L in lake and sea water samples, demonstrating that the proposed approach is an alternative means for rapid detection of pesticide residues and contaminants in food safety and environmental monitoring.
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