Threaded 3D microfluidic paper analytical device-based ratiometric fluorescent sensor for background-free and visual...

With the increasing concerns of food safety and environmental protection, it is desirable to develop reliable, effective, and portable sensors for detection of organophosphorus pesticides (OPs). Here, a cascade reaction system integrated with threaded 3D microfluidic paper analytical device (3D μPAD) was firstly developed for background-free and visual detection of OPs in agricultural samples. Butyrylcholinesterase (BChE) hydrolyzed acetylcholine into thiocholine (TCh), which reduced MnO₂ nanosheets into Mn²⁺. With addition of OPs, BChE activity was irreversibly inhibited, and the generation of TCh and the reduction of MnO₂ nanosheets were prevented. Then the remaining MnO₂ nanosheets oxidized o-phenylenediamine into 2,3-diaminophenazine with yellow-emission fluorescence, which quenched the fluorescence intensity of red-emission carbon dots (RCDs) via inner-filter effect. Based on above mechanism, a ratiometric fluorescent system was established for OPs detection. Threaded 3D μPAD consisted of 4 layers, which allowed to load and/or add reagents to trigger the cascade reaction system for OPs detection. The fluorescent images presented distinguishable color variations from red to yellow with dichlorvos concentrations ranging from 2.5 to 120 μg L^-1, and the limit of detection was 1.0 μg L^-1. In the practical samples testing, threaded 3D μPAD can eliminate background influence on fluorescent signal for OPs detection. Threaded 3D μPAD integrated with ratiometric sensing platform has merits of accuracy response, facile operation, and background-free detection, which supplies a new alternative approach for on-site pesticide detection.