Capillary-flow driven microfluidic sensor based on tyrosinase for fast user-friendly assessment of pesticide exposures

Pesticides are primarily used in agriculture to protect crops and extend their longevity. However, pesticide exposure has been linked to various acute and chronic health effects, raising significant environmental concerns. Current detection methods are often expensive and time-consuming, relying on complex instruments. Although enzyme-inhibition-based microfluidic paper-based analytical device (mPAD) platforms offer an easier alternative, they suffer from slow analyte transport and analyte adsorption issues in microchannels. Consequently, there is a need for a fast, simple, and cost-effective point-of-need platform for pesticide sensing. In this study, we present a rapid microfluidic platform for on-site pesticide residue detection. Unlike traditional mPAD platforms, our system transports pesticide samples through hollow capillary channels within seconds without adsorption of pesticides in the microchannels. While much research has focused on acetylcholinesterase inhibition on paper, this study is the first to introduce a tyrosinase inhibition-based assay on a paper platform for pesticide detection. Ziram, a representative dithiocarbamate pesticide, was detected using a colorimetric enzymatic inhibition assay. A limit of detection (LoD) of 1.5 ppm was obtained. In this study, we optimized the fast-flow device, assessed its stability and susceptibility to various interferences, and conducted real-sample tests using glove extraction to evaluate its capability in real-world settings. Spike recovery analysis revealed an extraction efficiency of 82.5% to 87.5% for leather gloves and 68.9% to 71.9% for nitrile gloves. This platform demonstrates strong selectivity against interferences, with the enzyme retaining 90% activity even after a week under the established storage protocols with room for further investigation. While primarily a proof of concept, this device shows promise as an additional tool for pesticide detection, with potential future integration into multiplexed devices. Assessment of pesticide exposure using a fast-flow capillary device (created with Bio.Render.com).