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J Agric Food Chem. 2019 Oct 9;67(40):11244-11255. doi: 10.1021/acs.jafc.9b03624. Epub 2019 Sep 27.

One-Step Facile Synthesis of Nitrogen-Doped Carbon Dots: A Ratiometric Fluorescent Probe for Evaluation of Acetylcholinesterase Activity and Detection of Organophosphorus Pesticides in Tap Water and Food.

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Guangxi Key Laboratory of Natural Polymer Chemistry and Physics, College of Chemistry and Materials , Nanning Normal University , Nanning 530001 , P. R. China.
Department of Food Science and Technology , University of Nebraska-Lincoln , Lincoln , Nebraska 68588 , United States.
School of Food Science and Biotechnology , Zhejiang Gongshang University , Hangzhou 310018 , P. R. China.


Evaluation of acetylcholinesterase (AChE) activity and determination of organophosphorus pesticides (OPs) are of great importance for the clinical diagnosis of several serious diseases correlated with their variations in human blood serum. In this study, a highly selective and sensitive ratiometric fluorescent probe was innovatively fabricated for the evaluation of AChE activity and the determination of OPs in tap water and food on the basis of the inner filter effect (IFE) between nitrogen-doped carbon dots (N-CDs) and 2,3-diaminophenazine (DAP). N-CDs were synthesized via a one-pot hydrothermal method by using pancreatin and 1,2-ethanediamine as precursors. N-CDs showed excellent fluorescence properties and negligible cytotoxicity on human cervical carcinoma HeLa cells and human embryonic kidney 293T cells, suggesting their further biological applications. Upon the addition of AChE and choline oxidase, acetylcholine was catalyzed to produce choline that was further oxidized to produce H2O2. In the presence of horseradish peroxidase, o-phenylenediamine reacted with H2O2 to produce fluorescent DAP. Therefore, a ratiometric fluorescent probing platform existed via IFE between N-CDs with a fluorescence signal at 450 nm and DAP with a fluorescence signal at 574 nm. OPs irreversibly impeded the catalytic activity of AChE, finally leading to the decrease of DAP amount and the variation of ratiometric fluorescent signal. Under optimal conditions, such a fluorescent probe showed relatively low detection limits of 0.38 U/L for AChE, 3.2 ppb for dichlorvos, and 13 ppb for methyl-parathion. Practical application of this ratiometric fluorescent probe to detect OPs was further verified in tap water and food samples with satisfying results that were highly consisted with the results obtained by GC-MS.


enzyme catalytic activity; inner filter effect; nitrogen-doped carbon dots; organophosphorus pesticides; ratiometric fluorescent probes


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