ORIGINAL RESEARCH article
Front. Pharmacol.
Sec. Ethnopharmacology
This article is part of the Research TopicNovel Technologies and Methods for Monitoring Exogenous Harmful Residues in Traditional and Local Medicinal Plants and FungiView all 5 articles
Electrostatically Driven Fluorescent Sensor for Rapid Detection of AChE activity and Organophosphate Pesticides via Dual-Enzyme Cascade Amplification
Provisionally accepted
Xie-an Yu*
Xiaoyi Liu
Kunhui Sun
Sitong LaiGuojing LiuMeifang LiPing WangShuhong Wang
Lan Ma
Bing Wang
Wei Lei- Shenzhen Institute For Drug Control, Shenzhen, China
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The widespread use of organophosphorus pesticides (OPs) in Chinese herbal medicines cultivation raises urgent concerns about residue contamination. Conventional detection methods (e.g., gas chromatography-mass spectrometry (GC-MS) and enzyme-linked immunosorbent assay (ELISA)) suffer from poor portability and instability of antibody inactivation in complex matrices, hindering on-site analysis. Here, this study proposed a novel "electrostatic adsorption-driven cascade reaction chain" strategy for rapid detection of acetylcholinesterase (AChE) activity and OPs. Leveraging the electrostatic self-assembly between a positively charged acetylcholine chloride (ACh, 26.47 ± 1.63 mV) and a negatively charged choline oxidase (CHO, –30.81 ± 1.85 mV), a nanoscale fluorescence sensor (CA-B NPs) was constructed by encapsulating the Azo-Bodipy 685. This design created a spatially confined and componentially co-localized nanoreactor that restricted substrate diffusion distance to the nanoscale and utilized a dual-enzyme cascade system (AChE-CHO) to yield a signal amplification effect. The obtained CA-B NPs exhibited excellent analytical performance, including: (1) a low detection limit of 4.1 ng/mL for triazophos; (2) high recovery of 88.13–113.09% in complex Citrus reticulata Blanco matrices, along with strong anti-interference capabilities by organically dividing the reaction and detection sections; (3) a total assay time of only 20 minutes for real samples, suitable for rapid, on-site, high-throughput screening. This study not only embedded the entire reaction chain (AChE-CHO-hydrogen peroxide (H2O2)) into the sensor to improve space utilization efficiency and detection efficiency, but also established a novel paradigm for enzyme spatial organization based on electrostatic complementarity, providing new insights into the rational design of nanostructured multi-enzyme sensing platforms.
Keywords: Azo-Bodipy 685, Fluorescent sensor, Acetylcholinesterase, Organophosphorus Pesticides, Electrostatic self-assembly
Received: 05 Aug 2025; Accepted: 18 Nov 2025.
Copyright: © 2025 Yu, Liu, Sun, Lai, Liu, Li, Wang, Wang, Ma, Wang and Lei. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Xie-an Yu, yuxieanalj@126.com
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